This application claims benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 63/276,868 filed Nov. 8, 2021 the contents of which are incorporated herein by reference in their entirety.

›GOVERNMENT SUPPORT

This invention was made with Government support under DK124361-01A1 and DK133404-01 awarded by the National Institute of Health. The Government has certain rights in the invention.

›SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on Jan. 6, 2023, is named 701586-191080USPX_SL.xml and is 106,377 bytes in size.

›TECHNICAL FIELD

The technology described herein relates to compositions and methods for engineered mRNAs encoding liver regeneration factors, e.g., engineered sequences and/or comprising modified nucleosides, to treat acute and chronic liver diseases.

›BACKGROUND

End stage liver disease (ESLD) is the 12th most common cause of death in the United States. As the result of a chronic damage of the liver tissue, ESLD begins as steatosis and inflammation and progresses to fibrosis and irreversible cirrhosis, and ultimately hepatocellular carcinoma. The current therapies to prevent the progression of the liver disease are designed to eliminate the underlying causes of injury including obesity or hepatitis C virus infection. Although some anti-fibrotic drugs are presently tested in clinical trials, none of them have been approved by the FDA. Liver transplantation remains the only treatment for ESLD, which is critically challenged by the shortage of liver donors. Currently, more than 6,000 liver transplants are performed each year in the United States, yet over 16,000 Americans are on the waiting list for a liver transplant. Given the scarcity of donor organs, hepatocyte transplantation has been attempted in patients with inherited metabolic liver and acute liver failure as treatment and a bridge for liver transplantation. Although the safety of the procedure is well established and the clinical results are encouraging, the application for liver cell therapy is still hampered by poor engraftment of transplanted cells, lack of optimal immunosuppression regiments and most importantly, a limited source of hepatocytes. The use of primary human hepatocytes could become a viable cell therapy for liver diseases only if cell engraftment mechanisms are significantly improved. Alternatively, hepatocytes derived from human induced pluripotent stem cells (hiPSC) could provide an unlimited supply for patient-specific cell replacement therapy. However, generation of iPSC-derived hepatocyte-like cells (HLC) that engraft, defined here as the ability to survive and proliferate, and are mature enough to function in a damaged liver remains a challenge, and a major gap.

Similarly, acute liver injury induced by overdose of acetaminophen (acetyl-para-aminophenol, APAP), the most common pain reliever consumed in the United States, is the leading cause of acute liver failure. Currently the only available treatment is the glutathione precursor N-acetyl cysteine (NAC), whose short window of effectiveness (˜10 h) ends while organ toxicity is frequently still asymptomatic, leading to liver failure and necessitating liver transplant. Alternative treatments for acute and chronic liver injuries including cell therapies and or therapies that would harness intrinsic liver repair mechanisms are therefore urgently needed.

›SUMMARY · 1 of 3

Chronic liver disease, acute liver disease, and acetaminophen (acetyl-para-aminophenol, APAP) overdose are major health burdens, for which improved treatments are needed. Uniquely, the liver is known for its remarkable regenerative ability through proliferation of hepatocytes. Therefore, an alternative strategy would be to promote regeneration of the injured liver tissues through endogenous mechanisms. Another strategy would be to improve primary hepatocyte and iPSC-derived hepatocyte-like cell transplantation and engraftment to take advantage of promising cell therapies.

The technology described herein is directed to compositions and methods of use of engineered liver regenerative factor mRNAs complexed to lipid nanoparticles (mRNA-LNP). This technology harnesses liver regeneration to accelerate intrinsic liver repair and to enhance cell therapy to ultimately treat both acute and chronic liver injuries. This technology also improves engraftment of primary human hepatocytes as well as induced pluripotent stem cell-derived hepatocyte-like cells generated from patient cells for successful cell therapy to treat acute and chronic liver diseases.

In one aspect of any of the embodiments, described herein is a composition comprising at least one engineered liver regenerative factor mRNA, the at least one engineered liver regenerative factor mRNA comprising one or more of:

a) a sequence encoding Growth Hormone (GH) and comprising one or more of the following modifications relative to SEQ ID NO: 1 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-63; T→C modification of nucleotide 69; A→C modification of nucleotide 72; G→C modification of nucleotide 81; G→C modification of nucleotide 84; C→G modification of nucleotide 92; T→C modification of nucleotide 99; T→C modification of nucleotide 102; C→G modification of nucleotide 111; T→G modification of nucleotide 126; A→G modification of nucleotide 129; A→T modification of nucleotide 136; G→C modification of nucleotide 137; T→C modification of nucleotide 138; A→C modification of nucleotide 147; T→C modification of nucleotide 153; T→C modification of nucleotide 157; A→G modification of nucleotide 159; A→C modification of nucleotide 163; G→C modification of nucleotide 165; T→G modification of nucleotide 168; T→C modification of nucleotide 171; T→C modification of nucleotide 180; C→G modification of nucleotide 186; T→C modification of nucleotide 195; T→C modification of nucleotide 198; T→C modification of nucleotide 216; T→C modification of nucleotide 234; A→G modification of nucleotide 237; A→G modification of nucleotide 240; T→C modification of nucleotide 246; A→C modification of nucleotide 252; A→G modification of nucleotide 258; T→C modification of nucleotide 267; A→C modification of nucleotide 270; C→G modification of nucleotide 297; T→C modification of nucleotide 300; A→C modification of nucleotide 306; T→C modification of nucleotide 312; T→C modification of nucleotide 315; G→C modification of nucleotide 318; A→C modification of nucleotide 321; A→C modification of nucleotide 331; G→C modification of nucleotide 333; A→G modification of nucleotide 339; A→C modification of nucleotide 342; A→G modification of nucleotide 345; A→G modification of nucleotide 351; A→G modification of nucleotide 360; C→G modification of nucleotide 369; C→G modification of nucleotide 387; G→C modification of nucleotide 396; C→G modification of nucleotide 420; A→C modification of nucleotide 421; G→C modification of nucleotide 423; A→T modification of nucleotide 424; G→C modification of nucleotide 425; T→C modification of nucleotide 426; C→G modification of nucleotide 429; A→T modification of nucleotide 439; G→C modification of nucleotide 440; T→C modification of nucleotide 459; A→T modification of nucleotide 463; G→C modification of nucleotide 464; C→G modification of nucleotide 471; T→C modification of nucleotide 474; C→G modification of nucleotide 480; A→G modification of nucleotide 483; A→G modification of nucleotide 492; A→G modification of nucleotide 498; A→G modification of nucleotide 507; G→C modification of nucleotide 510; G→C modification of nucleotide 519; A→C modification of nucleotide 520; G→C modification of nucleotide 522; A→G modification of nucleotide 528; T→C modification of nucleotide 531; A→T modification of nucleotide 535; G→C modification of nucleotide 536; G→C modification of nucleotide 543; T→C modification of nucleotide 546; G→C modification of nucleotide 549; A→T modification of nucleotide 571; G→C modification of nucleotide 572; A→C modification of nucleotide 585; A→C modification of nucleotide 591; T→C modification of nucleotide 600; A→C modification of nucleotide 606; A→G modification of nucleotide 609; C→G modification of nucleotide 612; G→C modification of nucleotide 624; C→G modification of nucleotide 630; A→C modification of nucleotide 640; G→C modification of nucleotide 642; C→G modification of nucleotide 660; A→C modification of nucleotide 666; T→C modification of nucleotide 693; A→T modification of nucleotide 703; G→C modification of nucleotide 704; T→C modification of nucleotide 708; and deletion of nucleotides 717-823; b) a sequence encoding Epidermal Growth Factor (EGF) and comprising one or more of the following modifications relative to SEQ ID NO: 2 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-453; C→G modification of nucleotide 462; T→C modification of nucleotide 465; T→G modification of nucleotide 468; T→C modification of nucleotide 474; T→C modification of nucleotide 478; A→C modification of nucleotide 483; A→G modification of nucleotide 486; T→G modification of nucleotide 489; A→C modification of nucleotide 492; A→G modification of nucleotide 495; T→C modification of nucleotide 498; A→T modification of nucleotide 499; G→C modification of nucleotide 500; T→C modification of nucleotide 501; T→C modification of nucleotide 504; T→G modification of nucleotide 507; A→T modification of nucleotide 508; G→C modification of nucleotide 509; T→C modification of nucleotide 510; G→C modification of nucleotide 513; A→C modification of nucleotide 516; A→C modification of nucleotide 519; G→C modification of nucleotide 522; A→T modification of nucleotide 532; G→C modification of nucleotide 533; T→C modification of nucleotide 537; T→C modification of nucleotide 540; A→G modification of nucleotide 543; T→C modification of nucleotide 546; T→C modification of nucleotide 549; C→G modification of nucleotide 552; A→C modification of nucleotide 555; A→C modification of nucleotide 558; T→C modification of nucleotide 561; G→C modification of nucleotide 564; T→C modification of nucleotide 567; T→C modification of nucleotide 570; T→C modification of nucleotide 573; T→C modification of nucleotide 576; T→C modification of nucleotide 582; T→C modification of nucleotide 585; A→C modification of nucleotide 588; T→C modification of nucleotide 595; A→G modification of nucleotide 597; T→C modification of nucleotide 600; T→C modification of nucleotide 609; A→C modification of nucleotide 612; T→C modification of nucleotide 615; A→T modification of nucleotide 616; G→C modification of nucleotide 617; T→C modification of nucleotide 618; T→C modification of nucleotide 624; A→C modification of nucleotide 625; G→C modification of nucleotide 627; T→C modification of nucleotide 630; A→C modification of nucleotide 636; A→G modification of nucleotide 639; A→C modification of nucleotide 642; T→C modification of nucleotide 648; T→C modification of nucleotide 651; A→G modification of nucleotide 657; T→C modification of nucleotide 658; T→C modification of nucleotide 669; T→C modification of nucleotide 672; T→C modification of nucleotide 675; C→G modification of nucleotide 678; A→C modification of nucleotide 681; T→C modification of nucleotide 693; T→C modification of nucleotide 696; T→C modification of nucleotide 699; T→C modification of nucleotide 702; T→C modification of nucleotide 705; A→G modification of nucleotide 711; A→C modification of nucleotide 712; A→C modification of nucleotide 714; T→C modification of nucleotide 720; T→C modification of nucleotide 729; T→C modification of nucleotide 730; A→G modification of nucleotide 732; A→G modification of nucleotide 735; A→C modification of nucleotide 736; A→C modification of nucleotide 738; A→G modification of nucleotide 741; T→G modification of nucleotide 744; T→C modification of nucleotide 745; A→G modification of nucleotide 750; A→C modification of nucleotide 751; A→C modification of nucleotide 753; T→G modification of nucleotide 756; T→C modification of nucleotide 759; T→C modification of nucleotide 765; G→C modification of nucleotide 768; A→C modification of nucleotide 771; A→C modification of nucleotide 772; G→C modification of nucleotide 774; A→G modification of nucleotide 777; A→C modification of nucleotide 781; A→C modification of nucleotide 783; A→G modification of nucleotide 786; T→C modification of nucleotide 789; T→C modification of nucleotide 792; A→C modification of nucleotide 795; A→G modification of nucleotide 801; T→C modification of nucleotide 804; T→G modification of nucleotide 807; T→C modification of nucleotide 810; A→C modification of nucleotide 813; A→C modification of nucleotide 819; A→C modification of nucleotide 822; T→C modification of nucleotide 825; A→C modification of nucleotide 831; T→C modification of nucleotide 834; A→G modification of nucleotide 837; A→G modification of nucleotide 840; T→G modification of nucleotide 843; T→C modification of nucleotide 846; A→C modification of nucleotide 852; T→C modification of nucleotide 855; A→G modification of nucleotide 858; A→G modification of nucleotide 864; A→C modification of nucleotide 867; T→C modification of nucleotide 873; A→C modification of nucleotide 876; A→G modification of nucleotide 879; A→C modification of nucleotide 882; T→C modification of nucleotide 885; A→G modification of nucleotide 891; A→C modification of nucleotide 894; T→C modification of nucleotide 897; T→C modification of nucleotide 900; T→C modification of nucleotide 909; T→G modification of nucleotide 912; T→C modification of nucleotide 913; A→G modification of nucleotide 915; A→T modification of nucleotide 916; G→C modification of nucleotide 917; T→C modification of nucleotide 918; T→C modification of nucleotide 921; T→C modification of nucleotide 922; A→G modification of nucleotide 924; A→G modification of nucleotide 927; T→C modification of nucleotide 930; T→C modification of nucleotide 933; A→C modification of nucleotide 936; T→C modification of nucleotide 939; A→G modification of nucleotide 942; A→C modification of nucleotide 945; T→G modification of nucleotide 948; T→C modification of nucleotide 951; A→C modification of nucleotide 954; A→G modification of nucleotide 957; A→G modification of nucleotide 960; A→C modification of nucleotide 961; T→C modification of nucleotide 966; A→C modification of nucleotide 972; T→C modification of nucleotide 978; A→C modification of nucleotide 981; T→C modification of nucleotide 990; A→C modification of nucleotide 993; A→T modification of nucleotide 994; G→C modification of nucleotide 995; T→G modification of nucleotide 999; T→C modification of nucleotide 1002; A→C modification of nucleotide 1003; A→C modification of nucleotide 1005; A→C modification of nucleotide 1008; T→C modification of nucleotide 1011; C→G modification of nucleotide 1014; T→C modification of nucleotide 1017; T→C modification of nucleotide 1020; A→C modification of nucleotide 1026; T→C modification of nucleotide 1035; T→C modification of nucleotide 1039; A→C modification of nucleotide 1047; A→C modification of nucleotide 1050; A→G modification of nucleotide 1056; A→C modification of nucleotide 1059; A→C modification of nucleotide 1062; T→C modification of nucleotide 1065; A→C modification of nucleotide 1071; T→C modification of nucleotide 1072; T→C modification of nucleotide 1077; T→G modification of nucleotide 1083; T→C modification of nucleotide 1086; G→C modification of nucleotide 1092; T→C modification of nucleotide 1098; T→C modification of nucleotide 1104; A→C modification of nucleotide 1114; A→C modification of nucleotide 1116; A→G modification of nucleotide 1119; A→C modification of nucleotide 1122; A→T modification of nucleotide 1123; G→C modification of nucleotide 1124; T→C modification of nucleotide 1128; T→C modification of nucleotide 1131; T→G modification of nucleotide 1134; T→C modification of nucleotide 1137; T→C modification of nucleotide 1146; T→C modification of nucleotide 1149; T→C modification of nucleotide 1152; T→C modification of nucleotide 1155; A→C modification of nucleotide 1158; T→C modification of nucleotide 1161; T→C modification of nucleotide 1164; C→G modification of nucleotide 1167; T→C modification of nucleotide 1173; A→T modification of nucleotide 1174; G→C modification of nucleotide 1175; T→C modification of nucleotide 1176; A→G modification of nucleotide 1179; T→C modification of nucleotide 1182; A→C modification of nucleotide 1185; A→C modification of nucleotide 1188; T→C modification of nucleotide 1194; T→C modification of nucleotide 1197; T→C modification of nucleotide 1198; T→C modification of nucleotide 1203; A→C modification of nucleotide 1206; T→G modification of nucleotide 1215; T→C modification of nucleotide 1218; T→C modification of nucleotide 1221; T→C modification of nucleotide 1227; T→C modification of nucleotide 1236; A→C modification of nucleotide 1239; A→C modification of nucleotide 1242; A→G modification of nucleotide 1248; A→C modification of nucleotide 1257; T→C modification of nucleotide 1260; A→C modification of nucleotide 1266; A→G modification of nucleotide 1275; T→C modification of nucleotide 1281; A→C modification of nucleotide 1284; T→G modification of nucleotide 1296; A→C modification of nucleotide 1297; A→C modification of nucleotide 1299; T→C modification of nucleotide 1302; C→G modification of nucleotide 1308; T→C modification of nucleotide 1311; A→C modification of nucleotide 1314; 1→C modification of nucleotide 1317; T→C modification of nucleotide 1320; A→G modification of nucleotide 1323; A→C modification of nucleotide 1326; T→G modification of nucleotide 1329; T→C modification of nucleotide 1332; A→G modification of nucleotide 1335; A→G modification of nucleotide 1341; A→G modification of nucleotide 1344; T→C modification of nucleotide 1350; A→C modification of nucleotide 1353; T→G modification of nucleotide 1356; A→C modification of nucleotide 1359; A→G modification of nucleotide 1362; A→C modification of nucleotide 1371; A→G modification of nucleotide 1374; T→C modification of nucleotide 1377; T→C modification of nucleotide 1383; T→C modification of nucleotide 1392; A→G modification of nucleotide 1401; T→G modification of nucleotide 1404; A→G modification of nucleotide 1410; A→C modification of nucleotide 1411; A→C modification of nucleotide 1414; G→C modification of nucleotide 1416; A→G modification of nucleotide 1419; A→C modification of nucleotide 1422; A→T modification of nucleotide 1429; G→C modification of nucleotide 1430; A→T modification of nucleotide 1432; G→C modification of nucleotide 1433; T→C modification of nucleotide 1437; T→C modification of nucleotide 1443; G→C modification of nucleotide 1446; A→G modification of nucleotide 1449; C→G modification of nucleotide 1455; A→C modification of nucleotide 1461; T→C modification of nucleotide 1465; T→C modification of nucleotide 1476; A→C modification of nucleotide 1479; A→C modification of nucleotide 1485; A→G modification of nucleotide 1494; A→T modification of nucleotide 1495; G→C modification of nucleotide 1496; T→C modification of nucleotide 1497; A→C modification of nucleotide 1500; G→C modification of nucleotide 1506; T→C modification of nucleotide 1515; A→G modification of nucleotide 1518; T→C modification of nucleotide 1521; T→G modification of nucleotide 1524; T→C modification of nucleotide 1527; A→G modification of nucleotide 1530; T→C modification of nucleotide 1536; T→C modification of nucleotide 1539; T→C modification of nucleotide 1545; T→C modification of nucleotide 1548; T→C modification of nucleotide 1554; T→C modification of nucleotide 1557; T→G modification of nucleotide 1560; G→C modification of nucleotide 1563; T→C modification of nucleotide 1566; A→G modification of nucleotide 1569; T→C modification of nucleotide 1578; A→C modification of nucleotide 1581; T→C modification of nucleotide 1587; G→C modification of nucleotide 1596; T→C modification of nucleotide 1602; A→G modification of nucleotide 1605; A→C modification of nucleotide 1608; T→C modification of nucleotide 1611; T→G modification of nucleotide 1614; T→G modification of nucleotide 1620; T→C modification of nucleotide 1623; T→C modification of nucleotide 1626; G→C modification of nucleotide 1629; A→G modification of nucleotide 1632; A→G modification of nucleotide 1635; T→C modification of nucleotide 1638; T→C modification of nucleotide 1641; A→G modification of nucleotide 1644; T→G modification of nucleotide 1647; T→G modification of nucleotide 1650; T→C modification of nucleotide 1656; A→C modification of nucleotide 1659; T→C modification of nucleotide 1665; T→C modification of nucleotide 1671; A→G modification of nucleotide 1674; A→T modification of nucleotide 1678; G→C modification of nucleotide 1679; T→C modification of nucleotide 1683; T→C modification of nucleotide 1689; T→G modification of nucleotide 1692; A→C modification of nucleotide 1698; A→C modification of nucleotide 1701; A→G modification of nucleotide 1704; T→C modification of nucleotide 1707; T→C modification of nucleotide 1711; A→G modification of nucleotide 1713; T→C modification of nucleotide 1716; T→C modification of nucleotide 1722; T→C modification of nucleotide 1725; A→G modification of nucleotide 1728; A→C modification of nucleotide 1734; T→G modification of nucleotide 1740; A→C modification of nucleotide 1744; A→C modification of nucleotide 1746; T→C modification of nucleotide 1749; G→C modification of nucleotide 1752; A→G modification of nucleotide 1755; A→C modification of nucleotide 1758; T→C modification of nucleotide 1761; A→T modification of nucleotide 1762; G→C modification of nucleotide 1763; T→C modification of nucleotide 1767; T→C modification of nucleotide 1770; A→C modification of nucleotide 1776; T→C modification of nucleotide 1782; T→C modification of nucleotide 1785; T→C modification of nucleotide 1788; A→C modification of nucleotide 1791; T→C modification of nucleotide 1794; A→T modification of nucleotide 1795; G→C modification of nucleotide 1796; C→G modification of nucleotide 1803; T→G modification of nucleotide 1809; T→C modification of nucleotide 1812; T→G modification of nucleotide 1815; A→T modification of nucleotide 1816; G→C modification of nucleotide 1817; A→C modification of nucleotide 1821; A→G modification of nucleotide 1824; A→G modification of nucleotide 1833; T→C modification of nucleotide 1836; T→C modification of nucleotide 1839; T→C modification of nucleotide 1845; T→C modification of nucleotide 1848; G→C modification of nucleotide 1851; T→C modification of nucleotide 1854; A→G modification of nucleotide 1860; A→G modification of nucleotide 1863; T→C modification of nucleotide 1869; A→G modification of nucleotide 1872; A→G modification of nucleotide 1875; G→C modification of nucleotide 1877; T→C modification of nucleotide 1881; A→C modification of nucleotide 1884; T→C modification of nucleotide 1887; A→C modification of nucleotide 1890; A→C modification of nucleotide 1893; A→C modification of nucleotide 1896; A→G modification of nucleotide 1899; A→C modification of nucleotide 1902; T→C modification of nucleotide 1905; T→C modification of nucleotide 1906; T→C modification of nucleotide 1914; T→C modification of nucleotide 1920; T→C modification of nucleotide 1923; A→G modification of nucleotide 1926; T→C modification of nucleotide 1929; T→C modification of nucleotide 1932; A→C modification of nucleotide 1935; T→C modification of nucleotide 1944; T→C modification of nucleotide 1947; T→C modification of nucleotide 1950; A→C modification of nucleotide 1953; A→C modification of nucleotide 1956; T→C modification of nucleotide 1962; A→C modification of nucleotide 1965; T→C modification of nucleotide 1968; C→G modification of nucleotide 1974; A→T modification of nucleotide 1975; G→C modification of nucleotide 1976; A→C modification of nucleotide 1989; T→G modification of nucleotide 1995; T→C modification of nucleotide 1998; A→G modification of nucleotide 2004; T→C modification of nucleotide 2007; T→C modification of nucleotide 2010; T→C modification of nucleotide 2016; A→G modification of nucleotide 2022; T→C modification of nucleotide 2025; A→C modification of nucleotide 2031; T→C modification of nucleotide 2037; T→C modification of nucleotide 2043; A→C modification of nucleotide 2046; A→C modification of nucleotide 2061; A→C modification of nucleotide 2065; A→C modification of nucleotide 2067; T→C modification of nucleotide 2070; T→C modification of nucleotide 2073; T→C modification of nucleotide 2079; T→C modification of nucleotide 2082; A→C modification of nucleotide 2091; A→G modification of nucleotide 2094; A→C modification of nucleotide 2095; G→C modification of nucleotide 2097; T→G modification of nucleotide 2100; T→C modification of nucleotide 2103; A→G modification of nucleotide 2109; A→C modification of nucleotide 2112; A→G modification of nucleotide 2115; T→C modification of nucleotide 2118; A→C modification of nucleotide 2124; A→G modification of nucleotide 2127; T→C modification of nucleotide 2130; T→G modification of nucleotide 2133; T→C modification of nucleotide 2136; T→C modification of nucleotide 2148; T→C modification of nucleotide 2154; A→C modification of nucleotide 2155; A→C modification of nucleotide 2157; T→C modification of nucleotide 2163; A→C modification of nucleotide 2169; A→C modification of nucleotide 2173; A→C modification of nucleotide 2175; G→C modification of nucleotide 2178; A→G modification of nucleotide 2181; T→C modification of nucleotide 2184; T→C modification of nucleotide 2190; A→C modification of nucleotide 2193; A→C modification of nucleotide 2194; G→C modification of nucleotide 2196; A→T modification of nucleotide 2197; G→C modification of nucleotide 2198; T→C modification of nucleotide 2199; T→C modification of nucleotide 2202; T→C modification of nucleotide 2203; A→G modification of nucleotide 2205; T→C modification of nucleotide 2208; G→C modification of nucleotide 2211; A→G modification of nucleotide 2214; T→G modification of nucleotide 2217; A→G modification of nucleotide 2223; A→C modification of nucleotide 2226; T→C modification of nucleotide 2232; T→C modification of nucleotide 2247; A→G modification of nucleotide 2250; A→C modification of nucleotide 2253; A→C modification of nucleotide 2256; A→C modification of nucleotide 2259; T→C modification of nucleotide 2262; T→C modification of nucleotide 2265; T→G modification of nucleotide 2268; T→C modification of nucleotide 2271; A→C modification of nucleotide 2274; A→C modification of nucleotide 2284; A→C modification of nucleotide 2286; T→C modification of nucleotide 2287; A→G modification of nucleotide 2289; T→C modification of nucleotide 2298; T→C modification of nucleotide 2301; A→C modification of nucleotide 2304; G→C modification of nucleotide 2307; T→C modification of nucleotide 2310; T→C modification of nucleotide 2313; A→C modification of nucleotide 2316; A→C modification of nucleotide 2319; T→C modification of nucleotide 2322; A→G modification of nucleotide 2325; A→T modification of nucleotide 2326; G→C modification of nucleotide 2327; T→C modification of nucleotide 2328; T→C modification of nucleotide 2331; C→G modification of nucleotide 2337; A→G modification of nucleotide 2340; T→G modification of nucleotide 2346; T→C modification of nucleotide 2352; T→G modification of nucleotide 2358; A→C modification of nucleotide 2361; T→A modification of nucleotide 2365; G→C modification of nucleotide 2366; T→C modification of nucleotide 2370; T→C modification of nucleotide 2373; A→G modification of nucleotide 2376; A→T modification of nucleotide 2386; G→C modification of nucleotide 2387; T→C modification of nucleotide 2388; A→C modification of nucleotide 2391; A→C modification of nucleotide 2394; G→C modification of nucleotide 2397; T→C modification of nucleotide 2400; T→C modification of nucleotide 2407; A→G modification of nucleotide 2409; T→C modification of nucleotide 2412; T→C modification of nucleotide 2419; T→C modification of nucleotide 2433; T→C modification of nucleotide 2445; T→C modification of nucleotide 2451; A→G modification of nucleotide 2454; T→C modification of nucleotide 2463; T→C modification of nucleotide 2469; T→C modification of nucleotide 2472; A→C modification of nucleotide 2475; A→G modification of nucleotide 2478; A→C modification of nucleotide 2484; A→C modification of nucleotide 2485; A→C modification of nucleotide 2487; T→G modification of nucleotide 2490; T→C modification of nucleotide 2499; T→C modification of nucleotide 2502; A→G modification of nucleotide 2505; T→C modification of nucleotide 2508; A→C modification of nucleotide 2514; T→C modification of nucleotide 2517; T→C modification of nucleotide 2520; A→G modification of nucleotide 2523; A→C modification of nucleotide 2526; T→C modification of nucleotide 2532; T→C modification of nucleotide 2538; T→C modification of nucleotide 2541; A→C modification of nucleotide 2553; T→C modification of nucleotide 2556; T→C modification of nucleotide 2562; A→C modification of nucleotide 2568; A→C modification of nucleotide 2571; A→G modification of nucleotide 2574; G→C modification of nucleotide 2577; A→C modification of nucleotide 2578; A→C modification of nucleotide 2580; A→G modification of nucleotide 2583; A→C modification of nucleotide 2590; G→C modification of nucleotide 2592; T→C modification of nucleotide 2595; A→G modification of nucleotide 2601; T→C modification of nucleotide 2604; A→C modification of nucleotide 2605; A→C modification of nucleotide 2607; A→G modification of nucleotide 2610; T→C modification of nucleotide 2613; A→C modification of nucleotide 2616; A→G modification of nucleotide 2619; A→T modification of nucleotide 2623; G→C modification of nucleotide 2624; A→C modification of nucleotide 2640; A→C modification of nucleotide 2643; T→G modification of nucleotide 2649; T→G modification of nucleotide 2655; T→C modification of nucleotide 2658; A→C modification of nucleotide 2661; T→C modification of nucleotide 2662; A→C modification of nucleotide 2667; A→G modification of nucleotide 2670; A→C modification of nucleotide 2673; A→C modification of nucleotide 2676; A→C modification of nucleotide 2679; T→C modification of nucleotide 2682; T→C modification of nucleotide 2689; A→G modification of nucleotide 2691; T→C modification of nucleotide 2694; A→G modification of nucleotide 2697; A→C modification of nucleotide 2703; T→C modification of nucleotide 2709; A→G modification of nucleotide 2712; T→C modification of nucleotide 2715; T→C modification of nucleotide 2718; A→G modification of nucleotide 2724; A→C modification of nucleotide 2728; G→C modification of nucleotide 2730; T→G modification of nucleotide 2733; A→C modification of nucleotide 2736; T→C modification of nucleotide 2739; T→C modification of nucleotide 2742; T→C modification of nucleotide 2748; G→C modification of nucleotide 2751; T→C modification of nucleotide 2754; T→C modification of nucleotide 2757; A→G modification of nucleotide 2760; T→C modification of nucleotide 2763; T→C modification of nucleotide 2766; A→G modification of nucleotide 2772; A→C modification of nucleotide 2778; T→C modification of nucleotide 2781; G→C modification of nucleotide 2784; A→G modification of nucleotide 2787; G→C modification of nucleotide 2790; T→C modification of nucleotide 2793; T→C modification of nucleotide 2799; T→C modification of nucleotide 2805; T→C modification of nucleotide 2808; T→C modification of nucleotide 2811; T→C modification of nucleotide 2818; A→C modification of nucleotide 2823; T→C modification of nucleotide 2826; T→C modification of nucleotide 2829; A→G modification of nucleotide 2832; T→G modification of nucleotide 2832; T→G modification of nucleotide 2835; T→C modification of nucleotide 2838; A→G modification of nucleotide 2841; A→G modification of nucleotide 2850; A→G modification of nucleotide 2853; A→C modification of nucleotide 2856; A→C modification of nucleotide 2859; T→C modification of nucleotide 2860; T→C modification of nucleotide 2869; T→C modification of nucleotide 2880; A→C modification of nucleotide 2881; A→C modification of nucleotide 2883; A→C modification of nucleotide 2889; A→G modification of nucleotide 2892; T→C modification of nucleotide 2895; T→C modification of nucleotide 2901; A→C modification of nucleotide 2904; A→G modification of nucleotide 2907; T→C modification of nucleotide 2910; A→G modification of nucleotide 2913; A→G modification of nucleotide 2922; T→C modification of nucleotide 2928; A→G modification of nucleotide 2931; A→C modification of nucleotide 2943; T→C modification of nucleotide 2946; A→G modification of nucleotide 2949; T→C modification of nucleotide 2952; T→C modification of nucleotide 2958; T→C modification of nucleotide 2961; T→C modification of nucleotide 2964; A→C modification of nucleotide 2970; A→T modification of nucleotide 2974; G→C modification of nucleotide 2975; T→C modification of nucleotide 2982; T→C modification of nucleotide 2985; G→C modification of nucleotide 2988; T→C modification of nucleotide 2991; T→C modification of nucleotide 2994; A→C modification of nucleotide 2997; A→C modification of nucleotide 3003; T→C modification of nucleotide 3009; A→C modification of nucleotide 3015; T→C modification of nucleotide 3018; T→C modification of nucleotide 3024; T→C modification of nucleotide 3025; A→G modification of nucleotide 3030; A→C modification of nucleotide 3033; T→C modification of nucleotide 3036; T→C modification of nucleotide 3039; G→C modification of nucleotide 3042; T→C modification of nucleotide 3045; A→C modification of nucleotide 3048; A→G modification of nucleotide 3051; A→G modification of nucleotide 3054; T→C modification of nucleotide 3057; T→C modification of nucleotide 3060; T→C modification of nucleotide 3063; A→C modification of nucleotide 3066; T→C modification of nucleotide 3069; A→G modification of nucleotide 3072; T→C modification of nucleotide 3075; T→C modification of nucleotide 3084; C→G modification of nucleotide 3087; A→C modification of nucleotide 3090; T→C modification of nucleotide 3102; A→G modification of nucleotide 3129; T→C modification of nucleotide 3132; T→C modification of nucleotide 3135; T→C modification of nucleotide 3138; C→G modification of nucleotide 3141; G→C modification of nucleotide 3147; A→C modification of nucleotide 3153; A→G modification of nucleotide 3156; A→G modification of nucleotide 3165; A→C modification of nucleotide 3168; T→C modification of nucleotide 3171; G→C modification of nucleotide 3174; T→C modification of nucleotide 3177; T→C modification of nucleotide 3183; T→G modification of nucleotide 3186; T→C modification of nucleotide 3189; T→C modification of nucleotide 3192; T→C modification of nucleotide 3195; A→G modification of nucleotide 3204; G→C modification of nucleotide 3210; A→T modification of nucleotide 3212; A→T modification of nucleotide 3217; G→C modification of nucleotide 3218; T→C modification of nucleotide 3222; A→C modification of nucleotide 3225; T→C modification of nucleotide 3231; A→T modification of nucleotide 3235; G→C modification of nucleotide 3236; A→C modification of nucleotide 3243; T→C modification of nucleotide 3246; A→C modification of nucleotide 3249; A→C modification of nucleotide 3255; T→C modification of nucleotide 3261; T→C modification of nucleotide 3273; T→C modification of nucleotide 3276; A→C modification of nucleotide 3279; T→C modification of nucleotide 3288; A→G modification of nucleotide 3291; A→C modification of nucleotide 3294; A→C modification of nucleotide 3297; T→C modification of nucleotide 3303; T→C modification of nucleotide 3309; T→C modification of nucleotide 3315; T→C modification of nucleotide 3318; A→C modification of nucleotide 3321; T→C modification of nucleotide 3327; C→G modification of nucleotide 3333; A→C modification of nucleotide 3334; G→C modification of nucleotide 3336; A→G modification of nucleotide 3339; T→C modification of nucleotide 3342; T→C modification of nucleotide 3354; A→G modification of nucleotide 3360; A→C modification of nucleotide 3361; A→C modification of nucleotide 3363; T→C modification of nucleotide 3366; T→A modification of nucleotide 3367; G→C modification of nucleotide 3368; T→C modification of nucleotide 3369; T→C modification of nucleotide 3375; A→G modification of nucleotide 3378; T→C modification of nucleotide 3381; T→C modification of nucleotide 3396; G→C modification of nucleotide 3399; C→G modification of nucleotide 3408; T→C modification of nucleotide 3411; T→C modification of nucleotide 3414; T→C modification of nucleotide 3417; T→C modification of nucleotide 3429; T→C modification of nucleotide 3432; A→G modification of nucleotide 3435; A→C modification of nucleotide 3438; T→C modification of nucleotide 3439; T→C modification of nucleotide 3450; A→C modification of nucleotide 3453; T→C modification of nucleotide 3462; T→G modification of nucleotide 3465; T→G modification of nucleotide 3468; G→C modification of nucleotide 3480; A→G modification of nucleotide 3486; T→C modification of nucleotide 3489; A→C modification of nucleotide 3498; A→G modification of nucleotide 3516; T→C modification of nucleotide 3528; G→C modification of nucleotide 3537; C→G modification of nucleotide 3552; T→C modification of nucleotide 3564; C→G modification of nucleotide 3567; T→G modification of nucleotide 3582; C→G modification of nucleotide 3585; C→G modification of nucleotide 3584; C→G modification of nucleotide 3594; C→G modification of nucleotide 3597; A→T modification of nucleotide 3612; G→C modification of nucleotide 3602; G→C modification of nucleotide 3612; A→C modification of nucleotide 3625; G→C modification of nucleotide 3627; T→C modification of nucleotide 3630; A→G modification of nucleotide 3642; G→C modification of nucleotide 3645; A→G modification of nucleotide 3648; A→C modification of nucleotide 3654; T→C modification of nucleotide 3660; T→C modification of nucleotide 3663; T→C modification of nucleotide 3668; G→C modification of nucleotide 3675; A→T modification of nucleotide 3676; G→C modification of nucleotide 3677; A→C modification of nucleotide 3679; A→C modification of nucleotide 3681; T→C modification of nucleotide 3684; A→C modification of nucleotide 3688; G→C modification of nucleotide 3690; A→T modification of nucleotide 3691; G→C modification of nucleotide 3692; T→C modification of nucleotide 3693; A→C modification of nucleotide 3697; G→C modification of nucleotide 3699; T→C modification of nucleotide 3702; T→C modification of nucleotide 3705; T→C modification of nucleotide 3711; T→C modification of nucleotide 3717; G→C modification of nucleotide 3720; T→C modification of nucleotide 3729; T→C modification of nucleotide 3735; A→G modification of nucleotide 3738; T→C modification of nucleotide 3741; T→C modification of nucleotide 3747; T→G modification of nucleotide 3752; A→C modification of nucleotide 3756; A→G modification of nucleotide 3759; A→G modification of nucleotide 3762; A→G modification of nucleotide 3769; C→G modification of nucleotide 3774; T→C modification of nucleotide 3780; G→C modification of nucleotide 3783; T→C modification of nucleotide 3786; A→G modification of nucleotide 3789; A→C modification of nucleotide 3792; T→C modification of nucleotide 3798; T→C modification of nucleotide 3801; T→C modification of nucleotide 3807; A→C modification of nucleotide 3816; A→C modification of nucleotide 3819; T→C modification of nucleotide 3822; G→C modification of nucleotide 3825; A→C modification of nucleotide 3828; A→G modification of nucleotide 3834; A→C modification of nucleotide 3837; T→C modification of nucleotide 3840; A→C modification of nucleotide 3843; A→C modification of nucleotide 3847; G→C modification of nucleotide 3849; T→C modification of nucleotide 3862; A→G modification of nucleotide 3864; T→C modification of nucleotide 3867; A→C modification of nucleotide 3870; A→C modification of nucleotide 3879; A→G modification of nucleotide 3885; T→C modification of nucleotide 3897; A→C modification of nucleotide 3900; A→G modification of nucleotide 3903; A→T modification of nucleotide 3907; G→C modification of nucleotide 3908; T→C modification of nucleotide 3909; T→C modification of nucleotide 3912; T→C modification of nucleotide 3924; A→G modification of nucleotide 3933; A→G modification of nucleotide 3942; A→T modification of nucleotide 3943; G→C modification of nucleotide 3944; T→C modification of nucleotide 3948; T→C modification of nucleotide 3951; T→C modification of nucleotide 3963; G→C modification of nucleotide 3966; A→C modification of nucleotide 3969; T→G modification of nucleotide 3978; A→G modification of nucleotide 3981; G→C modification of nucleotide 3984; T→C modification of nucleotide 3987; C→G modification of nucleotide 3990; T→C modification of nucleotide 4002; T→C modification of nucleotide 4005; C→G modification of nucleotide 4008; A→G modification of nucleotide 4011; A→C modification of nucleotide 4014; T→C modification of nucleotide 4017; A→C modification of nucleotide 4023; T→C modification of nucleotide 4024; A→G modification of nucleotide 4026; A→G modification of nucleotide 4032; A→G modification of nucleotide 4035; A→C modification of nucleotide 4036; G→C modification of nucleotide 4038; A→C modification of nucleotide 4050; A→C modification of nucleotide 4053; A→G modification of nucleotide 4059; T→C modification of nucleotide 4071; and deletion of nucleotides 4076-6388; c) a sequence encoding Hepatocyte Growth Factor (HGF) and comprising one or more of the following modifications relative to SEQ ID NO: 3 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-76; A→G modification of nucleotide 91; C→G modification of nucleotide 94; T→C modification of nucleotide 118; C→G modification of nucleotide 121; C→G modification of nucleotide 124; T→C modification of nucleotide 130; C→G modification of nucleotide 133; C→G modification of nucleotide 136; C→G modification of nucleotide 142; T→C modification of nucleotide 160; A→C modification of nucleotide 163; A→C modification of nucleotide 169; A→G modification of nucleotide 172; A→C modification of nucleotide 173; G→C modification of nucleotide 175; A→G modification of nucleotide 178; A→C modification of nucleotide 179; A→C modification of nucleotide 181; A→C modification of nucleotide 182; A→C modification of nucleotide 184; T→C modification of nucleotide 187; A→C modification of nucleotide 190; T→C modification of nucleotide 193; T→C modification of nucleotide 196; A→G modification of nucleotide 199; A→G modification of nucleotide 205; A→G modification of nucleotide 208; A→C modification of nucleotide 211; A→C modification of nucleotide 214; T→C modification of nucleotide 220; A→G modification of nucleotide 226; A→G modification of nucleotide 232; A→C modification of nucleotide 235; T→C modification of nucleotide 238; A→C modification of nucleotide 241; A→C modification of nucleotide 244; A→C modification of nucleotide 253; A→G modification of nucleotide 256; A→G modification of nucleotide 262; A→G modification of nucleotide 265; T→C modification of nucleotide 271; T→C modification of nucleotide 274; A→C modification of nucleotide 277; A→G modification of nucleotide 283; T→C modification of nucleotide 286; T→C modification of nucleotide 289; T→C modification of nucleotide 292; A→C modification of nucleotide 293; A→C modification of nucleotide 295; T→C modification of nucleotide 298; T→C modification of nucleotide 301; A→C modification of nucleotide 302; G→C modification of nucleotide 304; T→C modification of nucleotide 307; A→G modification of nucleotide 310; A→C modification of nucleotide 313; T→G modification of nucleotide 316; A→C modification of nucleotide 319; T→C modification of nucleotide 325; T→C modification of nucleotide 334; T→C modification of nucleotide 337; T→G modification of nucleotide 340; T→C modification of nucleotide 343; T→C modification of nucleotide 346; A→G modification of nucleotide 349; A→C modification of nucleotide 352; A→C modification of nucleotide 353; A→C modification of nucleotide 355; A→G modification of nucleotide 358; A→G modification of nucleotide 361; C→G modification of nucleotide 367; T→C modification of nucleotide 382; A→T modification of nucleotide 383; G→C modification of nucleotide 384; A→C modification of nucleotide 391; A→T modification of nucleotide 392; G→C modification of nucleotide 393; T→C modification of nucleotide 394; A→C modification of nucleotide 397; A→G modification of nucleotide 403; A→G modification of nucleotide 406; A→G modification of nucleotide 409; T→C modification of nucleotide 412; T→C modification of nucleotide 418; A→G modification of nucleotide 421; T→C modification of nucleotide 424; C→G modification of nucleotide 430; T→C modification of nucleotide 433; A→G modification of nucleotide 436; A→G modification of nucleotide 442; T→C modification of nucleotide 451; A→C modification of nucleotide 452; A→C modification of nucleotide 454; T→C modification of nucleotide 466; T→C modification of nucleotide 469; A→G modification of nucleotide 472; A→C modification of nucleotide 475; A→T modification of nucleotide 479; G→C modification of nucleotide 480; A→C modification of nucleotide 490; A→C modification of nucleotide 493; A→G modification of nucleotide 496; T→C modification of nucleotide 499; T→C modification of nucleotide 505; A→T modification of nucleotide 509; G→C modification of nucleotide 510; T→C modification of nucleotide 511; A→G modification of nucleotide 520; T→C modification of nucleotide 523; A→T modification of nucleotide 533; G→C modification of nucleotide 534; T→C modification of nucleotide 535; A→C modification of nucleotide 544; A→C modification of nucleotide 547; A→G modification of nucleotide 553; A→T modification of nucleotide 557; G→C modification of nucleotide 558; T→C modification of nucleotide 562; T→C modification of nucleotide 563; T→C modification of nucleotide 568; G→C modification of nucleotide 571; A→T modification of nucleotide 572; G→C modification of nucleotide 573; T→C modification of nucleotide 577; G→C modification of nucleotide 580; T→C modification of nucleotide 583; A→G modification of nucleotide 586; A→G modification of nucleotide 592; A→G modification of nucleotide 598; T→C modification of nucleotide 607; A→C modification of nucleotide 610; T→C modification of nucleotide 613; T→C modification of nucleotide 616; A→C modification of nucleotide 619; G→C modification of nucleotide 622; A→G modification of nucleotide 625; A→G modification of nucleotide 628; G→C modification of nucleotide 631; A→C modification of nucleotide 634; T→C modification of nucleotide 643; A→C modification of nucleotide 649; A→T modification of nucleotide 650; G→C modification of nucleotide 651; T→C modification of nucleotide 655; A→C modification of nucleotide 658; A→G modification of nucleotide 664; A→G modification of nucleotide 673; C→G modification of nucleotide 676; T→C modification of nucleotide 679; T→C modification of nucleotide 685; T→C modification of nucleotide 688; T→C modification of nucleotide 694; A→C modification of nucleotide 697; A→G modification of nucleotide 700; T→G modification of nucleotide 703; A→G modification of nucleotide 706; T→C modification of nucleotide 721; G→C modification of nucleotide 724; A→T modification of nucleotide 728; G→C modification of nucleotide 729; T→C modification of nucleotide 730; T→C modification of nucleotide 733; A→C modification of nucleotide 736; T→C modification of nucleotide 739; C→G modification of nucleotide 742; T→C modification of nucleotide 748; T→C modification of nucleotide 751; A→C modification of nucleotide 754; A→G modification of nucleotide 757; A→C modification of nucleotide 760; T→C modification of nucleotide 769; T→C modification of nucleotide 772; C→G modification of nucleotide 778; T→C modification of nucleotide 784; T→C modification of nucleotide 787; A→C modification of nucleotide 783; A→C modification of nucleotide 786; G→C modification of nucleotide 802; A→G modification of nucleotide 808; T→C modification of nucleotide 812; T→C modification of nucleotide 817; A→G modification of nucleotide 820; A→C modification of nucleotide 821; A→G modification of nucleotide 823; T→C modification of nucleotide 826; T→C modification of nucleotide 841; T→C modification of nucleotide 844; T→C modification of nucleotide 847; T→C modification of nucleotide 850; T→C modification of nucleotide 853; T→C modification of nucleotide 862; T→C modification of nucleotide 868; G→C modification of nucleotide 877; A→C modification of nucleotide 878; G→C modification of nucleotide 880; A→C modification of nucleotide 883; T→C modification of nucleotide 892; T→C modification of nucleotide 895; T→G modification of nucleotide 898; T→C modification of nucleotide 904; T→C modification of nucleotide 925; A→C modification of nucleotide 928; T→C modification of nucleotide 931; A→G modification of nucleotide 934; A→C modification of nucleotide 937; T→C modification of nucleotide 943; T→C modification of nucleotide 949; T→C modification of nucleotide 952; T→C modification of nucleotide 958; T→C modification of nucleotide 964; T→C modification of nucleotide 967; T→G modification of nucleotide 970; T→C modification of nucleotide 973; T→C modification of nucleotide 974; A→G modification of nucleotide 979; A→C modification of nucleotide 982; T→C modification of nucleotide 985; A→G modification of nucleotide 988; A→G modification of nucleotide 997; T→C modification of nucleotide 1000; A→G modification of nucleotide 1003; A→C modification of nucleotide 1006; A→G modification of nucleotide 1009; A→C modification of nucleotide 1016; G→C modification of nucleotide 1018; T→C modification of nucleotide 1024; C→G modification of nucleotide 1027; T→C modification of nucleotide 1030; T→C modification of nucleotide 1036; T→C modification of nucleotide 1042; A→C modification of nucleotide 1045; T→C modification of nucleotide 1048; A→C modification of nucleotide 1051; T→C modification of nucleotide 1054; T→G modification of nucleotide 1060; T→C modification of nucleotide 1066; T→C modification of nucleotide 1069; T→C modification of nucleotide 1075; T→C modification of nucleotide 1078; T→C modification of nucleotide 1087; T→C modification of nucleotide 1096; T→C modification of nucleotide 1099; A→G modification of nucleotide 1102; T→C modification of nucleotide 1105; A→G modification of nucleotide 1123; A→C modification of nucleotide 1126; A→G modification of nucleotide 1129; T→C modification of nucleotide 1132; A→C modification of nucleotide 1141; T→C modification of nucleotide 1144; A→C modification of nucleotide 1147; T→C modification of nucleotide 1150; G→C modification of nucleotide 1153; T→C modification of nucleotide 1156; A→G modification of nucleotide 1159; A→C modification of nucleotide 1162; T→C modification of nucleotide 1171; T→C modification of nucleotide 1174; T→C modification of nucleotide 1180; T→C modification of nucleotide 1183; A→C modification of nucleotide 1186; A→C modification of nucleotide 1195; T→G modification of nucleotide 1198; A→G modification of nucleotide 1213; T→C modification of nucleotide 1216; A→C modification of nucleotide 1219; T→C modification of nucleotide 1225; T→C modification of nucleotide 1228; A→C modification of nucleotide 1234; T→C modification of nucleotide 1237; A→C modification of nucleotide 1240; A→G modification of nucleotide 1243; T→C modification of nucleotide 1246; T→C modification of nucleotide 1249; T→C modification of nucleotide 1252; T→C modification of nucleotide 1255; G→C modification of nucleotide 1258; T→C modification of nucleotide 1261; A→G modification of nucleotide 1267; T→C modification of nucleotide 1270; T→C modification of nucleotide 1273; T→C modification of nucleotide 1283; A→G modification of nucleotide 1285; A→G modification of nucleotide 1291; A→C modification of nucleotide 1294; A→C modification of nucleotide 1295; A→C modification of nucleotide 1297; T→C modification of nucleotide 1300; A→C modification of nucleotide 1303; A→G modification of nucleotide 1306; A→C modification of nucleotide 1309; T→C modification of nucleotide 1312; A→C modification of nucleotide 1315; A→G modification of nucleotide 1336; T→C modification of nucleotide 1340; A→G modification of nucleotide 1342; T→C modification of nucleotide 1345; T→C modification of nucleotide 1348; T→C modification of nucleotide 1351; A→G modification of nucleotide 1363; A→C modification of nucleotide 1366; T→C modification of nucleotide 1369; A→C modification of nucleotide 1372; A→T modification of nucleotide 1373; G→C modification of nucleotide 1374; T→C modification of nucleotide 1375; T→C modification of nucleotide 1384; T→C modification of nucleotide 1390; A→C modification of nucleotide 1399; T→C modification of nucleotide 1402; A→C modification of nucleotide 1405; T→C modification of nucleotide 1408; T→C modification of nucleotide 1411; T→C modification of nucleotide 1414; T→C modification of nucleotide 1417; T→C modification of nucleotide 1420; A→C modification of nucleotide 1423; G→C modification of nucleotide 1438; A→C modification of nucleotide 1441; T→C modification of nucleotide 1444; A→C modification of nucleotide 1447; C→G modification of nucleotide 1450; T→C modification of nucleotide 1453; T→C modification of nucleotide 1456; T→C modification of nucleotide 1462; T→C modification of nucleotide 1465; T→C modification of nucleotide 1471; T→C modification of nucleotide 1474; T→C modification of nucleotide 1477; T→C modification of nucleotide 1480; T→C modification of nucleotide 1483; A→G modification of nucleotide 1486; T→C modification of nucleotide 1489; T→C modification of nucleotide 1492; A→C modification of nucleotide 1498; T→C modification of nucleotide 1501; A→C modification of nucleotide 1504; A→C modification of nucleotide 1507; C→G modification of nucleotide 1510; T→C modification of nucleotide 1513; T→C modification of nucleotide 1514; A→G modification of nucleotide 1516; T→C modification of nucleotide 1522; A→G modification of nucleotide 1528; A→C modification of nucleotide 1531; T→C modification of nucleotide 1534; T→C modification of nucleotide 1537; A→G modification of nucleotide 1543; G→C modification of nucleotide 1546; A→G modification of nucleotide 1549; A→G modification of nucleotide 1552; T→C modification of nucleotide 1553; A→C modification of nucleotide 1558; T→G modification of nucleotide 1561; A→G modification of nucleotide 1564; T→C modification of nucleotide 1567; G→C modification of nucleotide 1570; T→C modification of nucleotide 1573; A→C modification of nucleotide 1576; A→C modification of nucleotide 1579; A→C modification of nucleotide 1582; A→C modification of nucleotide 1585; A→C modification of nucleotide 1594; T→G modification of nucleotide 1603; A→T modification of nucleotide 1604; G→C modification of nucleotide 1605; T→C modification of nucleotide 1606; T→C modification of nucleotide 1607; A→C modification of nucleotide 1610; A→C modification of nucleotide 1612; A→C modification of nucleotide 1616; A→C modification of nucleotide 1618; T→C modification of nucleotide 1621; A→G modification of nucleotide 1624; T→C modification of nucleotide 1627; A→C modification of nucleotide 1636; A→C modification of nucleotide 1639; A→C modification of nucleotide 1642; T→C modification of nucleotide 1643; A→C modification of nucleotide 1648; A→T modification of nucleotide 1655; G→C modification of nucleotide 1656; T→C modification of nucleotide 1657; T→G modification of nucleotide 1663; T→G modification of nucleotide 1666; T→C modification of nucleotide 1669; A→C modification of nucleotide 1672; A→C modification of nucleotide 1675; T→C modification of nucleotide 1681; T→C modification of nucleotide 1687; T→C modification of nucleotide 1690; A→C modification of nucleotide 1693; T→C modification of nucleotide 1697; A→G modification of nucleotide 1702; T→C modification of nucleotide 1705; T→C modification of nucleotide 1708; A→G modification of nucleotide 1711; T→C modification of nucleotide 1714; T→G modification of nucleotide 1720; A→C modification of nucleotide 1723; T→C modification of nucleotide 1726; T→C modification of nucleotide 1729; T→C modification of nucleotide 1732; C→G modification of nucleotide 1735; A→C modification of nucleotide 1741; A→C modification of nucleotide 1742; A→C modification of nucleotide 1744; A→C modification of nucleotide 1747; T→C modification of nucleotide 1750; A→G modification of nucleotide 1756; A→G modification of nucleotide 1762; T→G modification of nucleotide 1768; C→G modification of nucleotide 1771; T→C modification of nucleotide 1774; T→G modification of nucleotide 1777; A→G modification of nucleotide 1789; T→C modification of nucleotide 1792; T→C modification of nucleotide 1798; A→G modification of nucleotide 1801; A→C modification of nucleotide 1804; A→C modification of nucleotide 1807; T→C modification of nucleotide 1810; T→G modification of nucleotide 1816; T→C modification of nucleotide 1817; A→G modification of nucleotide 1819; T→G modification of nucleotide 1828; A→C modification of nucleotide 1832; G→C modification of nucleotide 1834; T→C modification of nucleotide 1837; T→C modification of nucleotide 1840; C→G modification of nucleotide 1843; T→C modification of nucleotide 1849; T→C modification of nucleotide 1852; T→C modification of nucleotide 1855; T→G modification of nucleotide 1858; G→C modification of nucleotide 1860; T→C modification of nucleotide 1861; G→C modification of nucleotide 1863; T→C modification of nucleotide 1866; T→C modification of nucleotide 1870; T→C modification of nucleotide 1871; A→G modification of nucleotide 1873; T→C modification of nucleotide 1876; T→C modification of nucleotide 1879; T→C modification of nucleotide 1882; A→C modification of nucleotide 1885; A→C modification of nucleotide 1891; T→C modification of nucleotide 1894; T→C modification of nucleotide 1897; A→G modification of nucleotide 1900; A→T modification of nucleotide 1907; G→C modification of nucleotide 1908; T→C modification of nucleotide 1909; A→T modification of nucleotide 1913; G→C modification of nucleotide 1914; T→C modification of nucleotide 1915; T→G modification of nucleotide 1918; T→C modification of nucleotide 1921; T→C modification of nucleotide 1936; A→C modification of nucleotide 1939; T→C modification of nucleotide 1940; T→C modification of nucleotide 1951; T→C modification of nucleotide 1954; A→G modification of nucleotide 1960; T→C modification of nucleotide 1961; A→G modification of nucleotide 1963; A→C modification of nucleotide 1966; A→C modification of nucleotide 1972; T→C modification of nucleotide 1975; C→G modification of nucleotide 1978; T→C modification of nucleotide 1981; A→C modification of nucleotide 1990; T→C modification of nucleotide 1993; A→G modification of nucleotide 1999; A→T modification of nucleotide 2003; G→C modification of nucleotide 2004; T→C modification of nucleotide 2011; T→C modification of nucleotide 2014; A→C modification of nucleotide 2017; G→C modification of nucleotide 2020; T→C modification of nucleotide 2029; T→C modification of nucleotide 2035; T→C modification of nucleotide 2041; A→G modification of nucleotide 2044; A→C modification of nucleotide 2047; T→C modification of nucleotide 2050; T→C modification of nucleotide 2053; G→C modification of nucleotide 2056; T→C modification of nucleotide 2059; A→G modification of nucleotide 2062; T→C modification of nucleotide 2068; A→C modification of nucleotide 2071; A→C modification of nucleotide 2074; A→C modification of nucleotide 2077; A→C modification of nucleotide 2080; T→C modification of nucleotide 2083; G→C modification of nucleotide 2089; T→C modification of nucleotide 2092; T→C modification of nucleotide 2095; T→C modification of nucleotide 2098; A→C modification of nucleotide 2104; T→G modification of nucleotide 2107; T→G modification of nucleotide 2110; T→C modification of nucleotide 2113; A→G modification of nucleotide 2119; T→C modification of nucleotide 2122; A→G modification of nucleotide 2125; A→C modification of nucleotide 2129; A→C modification of nucleotide 2131; T→G modification of nucleotide 2137; T→G modification of nucleotide 2140; T→C modification of nucleotide 2143; C→G modification of nucleotide 2146; T→C modification of nucleotide 2149; T→G modification of nucleotide 2152; T→C modification of nucleotide 2155; T→C modification of nucleotide 2158; T→C modification of nucleotide 2161; A→C modification of nucleotide 2164; T→C modification of nucleotide 2167; T→C modification of nucleotide 2173; A→C modification of nucleotide 2176; T→C modification of nucleotide 2179; T→C modification of nucleotide 2182; T→C modification of nucleotide 2185; T→C modification of nucleotide 2188; T→C modification of nucleotide 2191; T→C modification of nucleotide 2194; C→G modification of nucleotide 2197; A→C modification of nucleotide 2200; A→G modification of nucleotide 2203; A→C modification of nucleotide 2206; T→C modification of nucleotide 2209; T→C modification of nucleotide 2212; A→C modification of nucleotide 2215; A→G modification of nucleotide 2218; A→C modification of nucleotide 2224; A→G modification of nucleotide 2230; T→C modification of nucleotide 2233; T→C modification of nucleotide 2236; T→C modification of nucleotide 2237; A→G modification of nucleotide 2239; A→C modification of nucleotide 2242; T→C modification of nucleotide 2245; A→G modification of nucleotide 2251; A→C modification of nucleotide 2254; A→C modification of nucleotide 2262; and deletion of nucleotides 2263-5834; d) a sequence encoding Cyclin-Dependent Kinase Inhibitor 1A (P21) and comprising one or more of the following modifications relative to SEQ ID NO: 4 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-134; T→C modification of nucleotide 143; T→C modification of nucleotide 146; T→C modification of nucleotide 149; T→C modification of nucleotide 152; C→G modification of nucleotide 155; A→C modification of nucleotide 158; T→C modification of nucleotide 161; T→G modification of nucleotide 164; G→C modification of nucleotide 167; A→C modification of nucleotide 171; G→C modification of nucleotide 173; A→T modification of nucleotide 174; G→C modification of nucleotide 175; A→G modification of nucleotide 179; T→C modification of nucleotide 188; T→C modification of nucleotide 191; C→G modification of nucleotide 194; T→C modification of nucleotide 200; A→T modification of nucleotide 210; G→C modification of nucleotide 211; T→C modification of nucleotide 212; T→C modification of nucleotide 219; T→C modification of nucleotide 227; T→C modification of nucleotide 230; T→C modification of nucleotide 236; G→C modification of nucleotide 239; C→G modification of nucleotide 242; G→C modification of nucleotide 248; T→C modification of nucleotide 254; C→G modification of nucleotide 257; A→C modification of nucleotide 269; A→G modification of nucleotide 272; T→C modification of nucleotide 284; C→G modification of nucleotide 293; G→C modification of nucleotide 296; G→C modification of nucleotide 302; G→C modification of nucleotide 305; C→G modification of nucleotide 323; T→G modification of nucleotide 335; G→C modification of nucleotide 338; A→T modification of nucleotide 339; G→C modification of nucleotide 340; A→G modification of nucleotide 344; G→C modification of nucleotide 347; C→G modification of nucleotide 359; A→T modification of nucleotide 366; G→C modification of nucleotide 367; T→C modification of nucleotide 371; G→C modification of nucleotide 374; A→T modification of nucleotide 381; G→C modification of nucleotide 382; T→C modification of nucleotide 386; A→C modification of nucleotide 398; G→C modification of nucleotide 401; A→C modification of nucleotide 408; G→C modification of nucleotide 410; A→T modification of nucleotide 414; G→C modification of nucleotide 415; T→C modification of nucleotide 416; T→C modification of nucleotide 419; T→C modification of nucleotide 425; G→C modification of nucleotide 440; A→C modification of nucleotide 443; T→C modification of nucleotide 446; G→C modification of nucleotide 449; T→C modification of nucleotide 465; G→C modification of nucleotide 470; T→C modification of nucleotide 476; T→C modification of nucleotide 482; T→C modification of nucleotide 491; G→C modification of nucleotide 497; T→C modification of nucleotide 500; A→G modification of nucleotide 503; T→C modification of nucleotide 506; G→C modification of nucleotide 512; T→C modification of nucleotide 515; G→C modification of nucleotide 518; A→C modification of nucleotide 524; A→C modification of nucleotide 527; T→C modification of nucleotide 530; A→C modification of nucleotide 539; A→G modification of nucleotide 542; G→C modification of nucleotide 545; A→C modification of nucleotide 546; G→C modification of nucleotide 548; A→G modification of nucleotide 555; G→C modification of nucleotide 556; A→C modification of nucleotide 563; T→C modification of nucleotide 566; T→C modification of nucleotide 572; A→C modification of nucleotide 585; A→C modification of nucleotide 587; T→C modification of nucleotide 588; C→G modification of nucleotide 593; A→C modification of nucleotide 603; A→C modification of nucleotide 605; A→G modification of nucleotide 608; and deletion of nucleotides 613-1943; e) a sequence encoding Vascular Endothelial Growth Factor A (VEGFA) and comprising one or more of the following modifications relative to SEQ ID NO: 5 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-1036; T→C modification of nucleotide 1045; T→C modification of nucleotide 1054; T→C modification of nucleotide 1063; A→T modification of nucleotide 1067; G→C modification of nucleotide 1068; T→G modification of nucleotide 1072; T→C modification of nucleotide 1076; C→G modification of nucleotide 1084; C→G modification of nucleotide 1090; T→C modification of nucleotide 1096; T→C modification of nucleotide 1114; A→C modification of nucleotide 1117; A→C modification of nucleotide 1126; A→G modification of nucleotide 1129; A→C modification of nucleotide 1132; A→C modification of nucleotide 1135; G→C modification of nucleotide 1138; T→C modification of nucleotide 1144; T→C modification of nucleotide 1147; A→G modification of nucleotide 1153; T→C modification of nucleotide 1171; C→G modification of nucleotide 1174; T→C modification of nucleotide 1177; C→G modification of nucleotide 1183; A→T modification of nucleotide 1184; G→C modification of nucleotide 1185; T→C modification of nucleotide 1195; A→C modification of nucleotide 1198; T→C modification of nucleotide 1234; T→C modification of nucleotide 1237; A→C modification of nucleotide 1261; T→C modification of nucleotide 1267; A→C modification of nucleotide 1282; G→C modification of nucleotide 1288; T→C modification of nucleotide 1300; T→C modification of nucleotide 1318; T→C modification of nucleotide 1327; T→C modification of nucleotide 1354; G→C modification of nucleotide 1360; A→G modification of nucleotide 1366; T→C modification of nucleotide 1369; A→G modification of nucleotide 1375; A→C modification of nucleotide 1387; A→C modification of nucleotide 1390; A→T modification of nucleotide 1397; G→C modification of nucleotide 1398; A→G modification of nucleotide 1405; A→G modification of nucleotide 1417; T→C modification of nucleotide 1420; A→G modification of nucleotide 1423; A→C modification of nucleotide 1427; A→C modification of nucleotide 1429; A→C modification of nucleotide 1432; A→G modification of nucleotide 1438; T→C modification of nucleotide 1441; A→C modification of nucleotide 1442; A→C modification of nucleotide 1444; A→C modification of nucleotide 1447; A→C modification of nucleotide 1448; A→C modification of nucleotide 1450; A→G modification of nucleotide 1453; A→G modification of nucleotide 1456; T→C modification of nucleotide 1459; T→C modification of nucleotide 1465; G→C modification of nucleotide 1468; T→C modification of nucleotide 1471; A→C modification of nucleotide 1477; G→C modification of nucleotide 1483; A→C modification of nucleotide 1484; A→C modification of nucleotide 1486; T→C modification of nucleotide 1492; T→C modification of nucleotide 1493; T→C modification of nucleotide 1498; A→G modification of nucleotide 1501; A→G modification of nucleotide 1504; T→C modification of nucleotide 1507; G→C modification of nucleotide 1510; G→C modification of nucleotide 1516; T→C modification of nucleotide 1519; A→G modification of nucleotide 1522; T→C modification of nucleotide 1525; A→G modification of nucleotide 1534; A→C modification of nucleotide 1540; G→C modification of nucleotide 1546; T→C modification of nucleotide 1549; and deletion of nucleotides 1556-3502; f) a sequence encoding Insulin-like Growth Factor 1 (Igf1) and comprising one or more of the following modifications relative to SEQ ID NO: 6 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-298; G→C modification of nucleotide 304; A→G modification of nucleotide 307; A→T modification of nucleotide 311; G→C modification of nucleotide 312; A→T modification of nucleotide 314; G→C modification of nucleotide 315; T→G modification of nucleotide 319; A→C modification of nucleotide 322; T→C modification of nucleotide 325; A→G modification of nucleotide 328; T→C modification of nucleotide 329; A→G modification of nucleotide 331; T→C modification of nucleotide 334; C→G modification of nucleotide 346; T→C modification of nucleotide 349; T→C modification of nucleotide 356; A→C modification of nucleotide 364; A→C modification of nucleotide 370; G→C modification of nucleotide 382; T→C modification of nucleotide 385; A→C modification of nucleotide 388; C→G modification of nucleotide 394; G→C modification of nucleotide 406; C→G modification of nucleotide 409; T→C modification of nucleotide 413; C→G modification of nucleotide 418; A→T modification of nucleotide 428; G→C modification of nucleotide 429; A→C modification of nucleotide 439; T→C modification of nucleotide 442; A→C modification of nucleotide 445; A→C modification of nucleotide 448; T→G modification of nucleotide 457; G→C modification of nucleotide 463; T→C modification of nucleotide 466; T→C modification of nucleotide 478; T→C modification of nucleotide 481; T→G modification of nucleotide 484; T→C modification of nucleotide 496; A→C modification of nucleotide 499; G→C modification of nucleotide 502; A→C modification of nucleotide 503; G→C modification of nucleotide 505; T→C modification of nucleotide 511; A→C modification of nucleotide 529; T→C modification of nucleotide 535; A→T modification of nucleotide 542; G→C modification of nucleotide 543; T→C modification of nucleotide 547; G→C modification of nucleotide 550; A→C modification of nucleotide 551; G→C modification of nucleotide 553; A→C modification of nucleotide 556; T→C modification of nucleotide 559; A→C modification of nucleotide 565; T→C modification of nucleotide 571; T→C modification of nucleotide 577; T→C modification of nucleotide 583; G→C modification of nucleotide 592; A→T modification of nucleotide 593; G→C modification of nucleotide 594; T→C modification of nucleotide 598; T→C modification of nucleotide 601; A→C modification of nucleotide 605; G→C modification of nucleotide 607; A→C modification of nucleotide 608; A→C modification of nucleotide 610; T→C modification of nucleotide 625; A→C modification of nucleotide 631; T→C modification of nucleotide 640; A→C modification of nucleotide 643; A→G modification of nucleotide 646; A→C modification of nucleotide 649; T→C modification of nucleotide 658; T→C modification of nucleotide 664; T→C modification of nucleotide 679; T→C modification of nucleotide 694; A→G modification of nucleotide 703; A→G modification of nucleotide 706; T→C modification of nucleotide 709; T→C modification of nucleotide 710; A→C modification of nucleotide 721; A→T modification of nucleotide 722; G→C modification of nucleotide 723; T→C modification of nucleotide 724; A→C modification of nucleotide 725; A→C modification of nucleotide 727; A→C modification of nucleotide 730; A→T modification of nucleotide 731; G→C modification of nucleotide 732; T→C modification of nucleotide 733; A→C modification of nucleotide 736; A→C modification of nucleotide 739; A→C modification of nucleotide 752; A→C modification of nucleotide 754; and deletion of nucleotides 760-7073; g) a sequence encoding Insulin-like Growth Factor 1 (Igf1) and comprising one or more of the following modifications relative to SEQ ID NO: 6 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-442; insertion of SEQ ID NO: 18; A→C modification of nucleotide 445; A→C modification of nucleotide 448; T→G modification of nucleotide 457; G→C modification of nucleotide 463; T→C modification of nucleotide 466; T→C modification of nucleotide 478; T→C modification of nucleotide 481; T→G modification of nucleotide 484; T→C modification of nucleotide 496; A→C modification of nucleotide 499; G→C modification of nucleotide 502; A→C modification of nucleotide 503; G→C modification of nucleotide 505; T→C modification of nucleotide 511; A→C modification of nucleotide 529; T→C modification of nucleotide 535; A→T modification of nucleotide 542; G→C modification of nucleotide 543; T→C modification of nucleotide 547; G→C modification of nucleotide 550; A→C modification of nucleotide 551; G→C modification of nucleotide 553; A→C modification of nucleotide 556; T→C modification of nucleotide 559; A→C modification of nucleotide 565; T→C modification of nucleotide 571; T→C modification of nucleotide 577; T→C modification of nucleotide 583; G→C modification of nucleotide 592; A→T modification of nucleotide 593; G→C modification of nucleotide 594; T→C modification of nucleotide 598; T→C modification of nucleotide 601; A→C modification of nucleotide 605; G→C modification of nucleotide 607; A→C modification of nucleotide 608; A→C modification of nucleotide 610; T→C modification of nucleotide 625; A→C modification of nucleotide 631; T→C modification of nucleotide 640; A→C modification of nucleotide 643; A→G modification of nucleotide 646; A→C modification of nucleotide 649; T→C modification of nucleotide 658; T→C modification of nucleotide 664; T→C modification of nucleotide 679; T→C modification of nucleotide 694; A→G modification of nucleotide 703; A→G modification of nucleotide 706; T→C modification of nucleotide 709; T→C modification of nucleotide 710; A→C modification of nucleotide 721; A→T modification of nucleotide 722; G→C modification of nucleotide 723; T→C modification of nucleotide 724; A→C modification of nucleotide 725; A→C modification of nucleotide 727; A→C modification of nucleotide 730; A→T modification of nucleotide 731; G→C modification of nucleotide 732; T→C modification of nucleotide 733; A→C modification of nucleotide 736; A→C modification of nucleotide 739; A→C modification of nucleotide 752; A→C modification of nucleotide 754; and deletion of nucleotides 760-7073; h) a sequence encoding Epidermal Growth Factor 1 (EGF) and comprising one or more of the following modifications relative to SEQ ID NO: 2 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-3369; insertion of SEQ ID NO: 19; T→C modification of nucleotide 3375; A→G modification of nucleotide 3378; T→C modification of nucleotide 3381; T→C modification of nucleotide 3396; G→C modification of nucleotide 3399; C→G modification of nucleotide 3408; T→C modification of nucleotide 3411; T→C modification of nucleotide 3414; T→C modification of nucleotide 3417; T→C modification of nucleotide 3429; T→C modification of nucleotide 3432; A→G modification of nucleotide 3435; A→C modification of nucleotide 3438; T→C modification of nucleotide 3439; T→C modification of nucleotide 3450; A→C modification of nucleotide 3453; T→C modification of nucleotide 3462; T→G modification of nucleotide 3465; T→G modification of nucleotide 3468; G→C modification of nucleotide 3480; A→G modification of nucleotide 3486; T→C modification of nucleotide 3489; A→C modification of nucleotide 3498; A→G modification of nucleotide 3516; and deletion of nucleotides 3523-6388; i) a sequence encoding signal transducer and activator of transcription 5B (Stat5b) and comprising one or more of the following modifications relative to SEQ ID NO: 7 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-532; insertion of SEQ ID NO: 20; T→C modification of nucleotide 535; A→C modification of nucleotide 544; T→C modification of nucleotide 550; C→G modification of nucleotide 559; T→C modification of nucleotide 568; T→G modification of nucleotide 574; T→C modification of nucleotide 590; T→C modification of nucleotide 604; A→C modification of nucleotide 622; T→C modification of nucleotide 625; T→C modification of nucleotide 628; T→C modification of nucleotide 629; A→G modification of nucleotide 631; A→C modification of nucleotide 634; A→G modification of nucleotide 646; A→T modification of nucleotide 647; G→C modification of nucleotide 648; A→G modification of nucleotide 652; A→C modification of nucleotide 664; A→C modification of nucleotide 667; T→C modification of nucleotide 670; T→G modification of nucleotide 673; T→C modification of nucleotide 676; T→C modification of nucleotide 679; A→C modification of nucleotide 682; T→C modification of nucleotide 694; C→G modification of nucleotide 709; G→C modification of nucleotide 745; G→C modification of nucleotide 760; A→G modification of nucleotide 763; T→C modification of nucleotide 766; G→C modification of nucleotide 769; T→C modification of nucleotide 772; T→C modification of nucleotide 773; G→C modification of nucleotide 793; T→C modification of nucleotide 799; A→C modification of nucleotide 805; C→G modification of nucleotide 811; A→T modification of nucleotide 815; G→C modification of nucleotide 816; G→C modification of nucleotide 820; T→G modification of nucleotide 847; T→C modification of nucleotide 853; G→C modification of nucleotide 859; T→C modification of nucleotide 865; A→G modification of nucleotide 877; A→C modification of nucleotide 881; G→C modification of nucleotide 883; T→G modification of nucleotide 889; A→G modification of nucleotide 895; A→T modification of nucleotide 908; G→C modification of nucleotide 909; T→C modification of nucleotide 913; A→C modification of nucleotide 916; T→C modification of nucleotide 919; A→C modification of nucleotide 922; A→T modification of nucleotide 923; G→C modification of nucleotide 924; T→C modification of nucleotide 925; T→G modification of nucleotide 928; T→C modification of nucleotide 931; T→G modification of nucleotide 955; A→G modification of nucleotide 967; G→C modification of nucleotide 970; T→C modification of nucleotide 973; A→C modification of nucleotide 994; G→C modification of nucleotide 1003; A→G modification of nucleotide 1036; A→T modification of nucleotide 1064; G→C modification of nucleotide 1065; G→C modification of nucleotide 1072; A→G modification of nucleotide 1078; T→C modification of nucleotide 1081; T→C modification of nucleotide 1087; A→C modification of nucleotide 1099; A→T modification of nucleotide 1124; G→C modification of nucleotide 1125; A→C modification of nucleotide 1127; G→C modification of nucleotide 1129; G→C modification of nucleotide 1135; C→G modification of nucleotide 1141; A→G modification of nucleotide 1153; A→C modification of nucleotide 1180; A→C modification of nucleotide 1186; A→C modification of nucleotide 1192; A→C modification of nucleotide 1207; T→C modification of nucleotide 1219; G→C modification of nucleotide 1252; G→C modification of nucleotide 1300; A→C modification of nucleotide 1301; A→C modification of nucleotide 1303; G→C modification of nucleotide 1318; G→C modification of nucleotide 1324; T→C modification of nucleotide 1327; A→T modification of nucleotide 1340; G→C modification of nucleotide 1341; T→C modification of nucleotide 1366; G→C modification of nucleotide 1399; A→C modification of nucleotide 1412; G→C modification of nucleotide 1414; T→C modification of nucleotide 1417; T→C modification of nucleotide 1424; A→C modification of nucleotide 1447; T→C modification of nucleotide 1471; C→G modification of nucleotide 1477; G→C modification of nucleotide 1492; A→C modification of nucleotide 1504; C→G modification of nucleotide 1513; A→T modification of nucleotide 1517; G→C modification of nucleotide 1518; G→C modification of nucleotide 1522; T→C modification of nucleotide 1543; T→C modification of nucleotide 1546; C→G modification of nucleotide 1552; T→C modification of nucleotide 1573; A→C modification of nucleotide 1576; G→C modification of nucleotide 1600; G→C modification of nucleotide 1603; T→C modification of nucleotide 1612; G→C modification of nucleotide 1630; G→C modification of nucleotide 1642; A→T modification of nucleotide 1652; G→C modification of nucleotide 1653; C→G modification of nucleotide 1678; T→C modification of nucleotide 1684; T→C modification of nucleotide 1699; T→C modification of nucleotide 1702; A→T modification of nucleotide 1706; G→C modification of nucleotide 1707; T→C modification of nucleotide 1729; C→G modification of nucleotide 1735; T→C modification of nucleotide 1756; A→C modification of nucleotide 1762; C→G modification of nucleotide 1765; A→T modification of nucleotide 1766; G→C modification of nucleotide 1767; A→C modification of nucleotide 1778; A→C modification of nucleotide 1780; A→G modification of nucleotide 1795; A→C modification of nucleotide 1798; A→C modification of nucleotide 1805; T→C modification of nucleotide 1810; T→C modification of nucleotide 1819; G→C modification of nucleotide 1822; A→C modification of nucleotide 1825; A→C modification of nucleotide 1831; A→G modification of nucleotide 1834; G→C modification of nucleotide 1837; A→G modification of nucleotide 1840; G→C modification of nucleotide 1852; T→C modification of nucleotide 1861; A→C modification of nucleotide 1867; A→T modification of nucleotide 1874; G→C modification of nucleotide 1875; C→G modification of nucleotide 1879; T→C modification of nucleotide 1882; A→C modification of nucleotide 1885; C→G modification of nucleotide 1897; T→C modification of nucleotide 1900; A→G modification of nucleotide 1903; C→G modification of nucleotide 1906; T→C modification of nucleotide 1913; G→C modification of nucleotide 1918; C→G modification of nucleotide 1921; G→C modification of nucleotide 1924; T→C modification of nucleotide 1936; T→G modification of nucleotide 1939; A→T modification of nucleotide 1946; G→C modification of nucleotide 1947; T→C modification of nucleotide 1960; A→C modification of nucleotide 1966; T→C modification of nucleotide 1972; C→G modification of nucleotide 1975; C→G modification of nucleotide 1978; T→C modification of nucleotide 1993; A→C modification of nucleotide 1996; T→C modification of nucleotide 2002; A→C modification of nucleotide 2006; G→C modification of nucleotide 2008; A→C modification of nucleotide 2014; T→C modification of nucleotide 2017; T→C modification of nucleotide 2026; G→C modification of nucleotide 2044; T→C modification of nucleotide 2053; A→G modification of nucleotide 2056; G→C modification of nucleotide 2059; C→G modification of nucleotide 2062; A→G modification of nucleotide 2071; T→C modification of nucleotide 2080; A→G modification of nucleotide 2083; A→G modification of nucleotide 2086; A→T modification of nucleotide 2090; G→C modification of nucleotide 2091; G→C modification of nucleotide 2098; T→C modification of nucleotide 2102; C→G modification of nucleotide 2119; A→C modification of nucleotide 2131; A→G modification of nucleotide 2137; A→T modification of nucleotide 2150; G→C modification of nucleotide 2151; A→T modification of nucleotide 2153; G→C modification of nucleotide 2154; C→G modification of nucleotide 2164; A→T modification of nucleotide 2177; G→C modification of nucleotide 2178; G→C modification of nucleotide 2209; T→C modification of nucleotide 2215; T→C modification of nucleotide 2216; A→C modification of nucleotide 2221; A→C modification of nucleotide 2224; G→C modification of nucleotide 2227; T→C modification of nucleotide 2230; T→C modification of nucleotide 2236; T→C modification of nucleotide 2251; T→C modification of nucleotide 2254; A→G modification of nucleotide 2266; A→G modification of nucleotide 2269; T→C modification of nucleotide 2270; A→G modification of nucleotide 2275; A→G modification of nucleotide 2278; T→C modification of nucleotide 2281; C→G modification of nucleotide 2284; T→C modification of nucleotide 2290; T→C modification of nucleotide 2299; T→C modification of nucleotide 2302; G→C modification of nucleotide 2305; T→C modification of nucleotide 2308; T→C modification of nucleotide 2317; A→G modification of nucleotide 2332; C→G modification of nucleotide 2350; A→C modification of nucleotide 2362; G→C modification of nucleotide 2368; A→T modification of nucleotide 2387; G→C modification of nucleotide 2388; G→C modification of nucleotide 2395; A→G modification of nucleotide 2398; G→C modification of nucleotide 2404; T→C modification of nucleotide 2416; T→C modification of nucleotide 2419; T→C modification of nucleotide 2428; T→C modification of nucleotide 2434; A→C modification of nucleotide 2441; A→C modification of nucleotide 2443; T→C modification of nucleotide 2449; A→C modification of nucleotide 2453; T→C modification of nucleotide 2455; T→C modification of nucleotide 2464; T→C modification of nucleotide 2467; T→C modification of nucleotide 2473; A→C modification of nucleotide 2474; A→C modification of nucleotide 2476; T→C modification of nucleotide 2485; G→C modification of nucleotide 2491; C→G modification of nucleotide 2497; T→C modification of nucleotide 2500; G→C modification of nucleotide 2512; T→C modification of nucleotide 2521; C→G modification of nucleotide 2527; A→C modification of nucleotide 2530; T→C modification of nucleotide 2533; T→C modification of nucleotide 2539; T→C modification of nucleotide 2542; T→C modification of nucleotide 2545; G→C modification of nucleotide 2548; A→C modification of nucleotide 2551; T→C modification of nucleotide 2557; A→G modification of nucleotide 2560; A→G modification of nucleotide 2563; T→C modification of nucleotide 2566; T→C modification of nucleotide 2569; A→C modification of nucleotide 2581; G→C modification of nucleotide 2584; C→G modification of nucleotide 2587; T→C modification of nucleotide 2593; A→C modification of nucleotide 2602; T→C modification of nucleotide 2605; G→C modification of nucleotide 2608; A→G modification of nucleotide 2611; A→C modification of nucleotide 2614; T→C modification of nucleotide 2617; A→C modification of nucleotide 2623; A→C modification of nucleotide 2635; C→G modification of nucleotide 2653; T→C modification of nucleotide 2662; A→C modification of nucleotide 2665; T→C modification of nucleotide 2668; A→C modification of nucleotide 2671; A→C modification of nucleotide 2677; T→C modification of nucleotide 2680; T→C modification of nucleotide 2683; G→C modification of nucleotide 2686; A→T modification of nucleotide 2687; G→C modification of nucleotide 2688; T→C modification of nucleotide 2689; T→C modification of nucleotide 2707; T→C modification of nucleotide 2713; T→C modification of nucleotide 2716; A→C modification of nucleotide 2722; C→G modification of nucleotide 2725; T→C modification of nucleotide 2734; T→C modification of nucleotide 2740; A→C modification of nucleotide 2758; G→C modification of nucleotide 2767; C→G modification of nucleotide 2776; T→G modification of nucleotide 2779; T→C modification of nucleotide 2782; T→C modification of nucleotide 2788; G→C modification of nucleotide 2791; T→C modification of nucleotide 2800; A→G modification of nucleotide 2806; G→C modification of nucleotide 2812; G→C modification of nucleotide 2824; G→C modification of nucleotide 2827; A→G modification of nucleotide 2836; C→G modification of nucleotide 2842; T→C modification of nucleotide 2843; A→G modification of nucleotide 2845; G→C modification of nucleotide 2851; A→T modification of nucleotide 2861; G→C modification of nucleotide 2862; T→C modification of nucleotide 2863; T→C modification of nucleotide 2875; A→C modification of nucleotide 2881; A→C modification of nucleotide 2887; and deletion of nucleotides 2888-5255; j) a sequence encoding beta catenin (CTNNB1) and comprising one or more of the following modifications relative to SEQ ID NO: 17 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-214; T→C modification of nucleotide 220; T→C modification of nucleotide 223; A→G modification of nucleotide 226; T→C modification of nucleotide 229; T→C modification of nucleotide 232; T→C modification of nucleotide 233; T→C modification of nucleotide 242; A→G modification of nucleotide 259; A→C modification of nucleotide 262; A→C modification of nucleotide 266; A→C modification of nucleotide 268; A→G modification of nucleotide 271; G→C modification of nucleotide 274; T→C modification of nucleotide 277; T→G modification of nucleotide 280; A→T modification of nucleotide 281; G→C modification of nucleotide 282; T→C modification of nucleotide 283; A→G modification of nucleotide 295; T→C modification of nucleotide 301; T→C modification of nucleotide 313; A→C modification of nucleotide 316; T→C modification of nucleotide 322; T→C modification of nucleotide 325; T→C modification of nucleotide 328; T→C modification of nucleotide 334; A→C modification of nucleotide 340; T→C modification of nucleotide 343; T→C modification of nucleotide 346; T→C modification of nucleotide 349; A→C modification of nucleotide 353; G→C modification of nucleotide 354; T→C modification of nucleotide 355; T→C modification of nucleotide 358; A→G modification of nucleotide 361; T→C modification of nucleotide 367; T→C modification of nucleotide 370; A→G modification of nucleotide 376; T→C modification of nucleotide 382; T→C modification of nucleotide 388; A→G modification of nucleotide 397; C→G modification of nucleotide 400; T→C modification of nucleotide 406; A→G modification of nucleotide 415; A→C modification of nucleotide 421; T→C modification of nucleotide 424; T→C modification of nucleotide 427; T→C modification of nucleotide 439; A→G modification of nucleotide 442; A→G modification of nucleotide 445; A→G modification of nucleotide 448; A→G modification of nucleotide 451; T→C modification of nucleotide 454; T→C modification of nucleotide 457; T→C modification of nucleotide 460; T→C modification of nucleotide 463; A→C modification of nucleotide 466; T→C modification of nucleotide 472; A→C modification of nucleotide 475; T→C modification of nucleotide 481; A→C modification of nucleotide 484; T→C modification of nucleotide 487; A→C modification of nucleotide 491; G→C modification of nucleotide 493; A→G modification of nucleotide 496; A→C modification of nucleotide 499; T→C modification of nucleotide 502; T→C modification of nucleotide 505; T→C modification of nucleotide 514; A→C modification of nucleotide 520; T→C modification of nucleotide 521; A→G modification of nucleotide 523; T→C modification of nucleotide 526; A→C modification of nucleotide 544; T→C modification of nucleotide 547; A→C modification of nucleotide 550; T→C modification of nucleotide 556; T→C modification of nucleotide 559; T→C modification of nucleotide 562; T→C modification of nucleotide 565; T→C modification of nucleotide 568; T→C modification of nucleotide 574; T→C modification of nucleotide 577; C→G modification of nucleotide 580; T→C modification of nucleotide 586; T→C modification of nucleotide 587; T→C modification of nucleotide 592; A→G modification of nucleotide 595; A→C modification of nucleotide 598; A→C modification of nucleotide 601; A→G modification of nucleotide 613; T→C modification of nucleotide 616; A→C modification of nucleotide 619; T→G modification of nucleotide 622; A→G modification of nucleotide 625; T→C modification of nucleotide 629; T→C modification of nucleotide 634; T→C modification of nucleotide 640; A→G modification of nucleotide 643; T→C modification of nucleotide 646; T→C modification of nucleotide 649; A→C modification of nucleotide 652; A→G modification of nucleotide 655; T→G modification of nucleotide 658; A→C modification of nucleotide 664; T→C modification of nucleotide 667; A→C modification of nucleotide 670; T→C modification of nucleotide 676; A→G modification of nucleotide 679; A→C modification of nucleotide 685; A→G modification of nucleotide 688; A→G modification of nucleotide 694; T→C modification of nucleotide 697; T→G modification of nucleotide 718; T→C modification of nucleotide 721; T→C modification of nucleotide 727; A→C modification of nucleotide 730; T→G modification of nucleotide 733; C→G modification of nucleotide 739; T→C modification of nucleotide 742; T→G modification of nucleotide 748; T→C modification of nucleotide 751; A→G modification of nucleotide 754; A→G modification of nucleotide 760; T→C modification of nucleotide 763; A→C modification of nucleotide 767; A→C modification of nucleotide 769; T→C modification of nucleotide 775; T→C modification of nucleotide 784; T→C modification of nucleotide 787; T→C modification of nucleotide 790; T→C modification of nucleotide 802; T→C modification of nucleotide 805; T→C modification of nucleotide 808; A→G modification of nucleotide 811; T→C modification of nucleotide 814; T→C modification of nucleotide 826; A→C modification of nucleotide 829; T→C modification of nucleotide 832; T→C modification of nucleotide 835; A→G modification of nucleotide 838; A→G modification of nucleotide 841; A→C modification of nucleotide 844; T→C modification of nucleotide 847; T→C modification of nucleotide 850; T→C modification of nucleotide 853; T→C modification of nucleotide 859; G→C modification of nucleotide 862; T→C modification of nucleotide 866; T→C modification of nucleotide 871; T→G modification of nucleotide 878; T→C modification of nucleotide 883; T→C modification of nucleotide 886; T→C modification of nucleotide 889; T→C modification of nucleotide 896; A→G modification of nucleotide 898; T→C modification of nucleotide 910; T→C modification of nucleotide 916; A→C modification of nucleotide 919; T→C modification of nucleotide 925; T→C modification of nucleotide 928; A→G modification of nucleotide 940; T→G modification of nucleotide 946; T→C modification of nucleotide 949; A→C modification of nucleotide 952; A→C modification of nucleotide 955; T→C modification of nucleotide 961; T→C modification of nucleotide 964; T→C modification of nucleotide 968; T→C modification of nucleotide 973; T→C modification of nucleotide 976; T→C modification of nucleotide 982; A→C modification of nucleotide 985; T→C modification of nucleotide 988; C→G modification of nucleotide 991; T→G modification of nucleotide 1000; T→C modification of nucleotide 1001; A→G modification of nucleotide 1003; T→C modification of nucleotide 1004; A→G modification of nucleotide 1006; T→C modification of nucleotide 1009; A→G modification of nucleotide 1012; A→G modification of nucleotide 1015; A→C modification of nucleotide 1018; T→C modification of nucleotide 1021; A→G modification of nucleotide 1024; A→C modification of nucleotide 1030; T→C modification of nucleotide 1036; T→C modification of nucleotide 1037; A→G modification of nucleotide 1039; T→C modification of nucleotide 1042; T→C modification of nucleotide 1045; G→C modification of nucleotide 1048; A→G modification of nucleotide 1057; T→G modification of nucleotide 1063; T→C modification of nucleotide 1067; C→G modification of nucleotide 1072; A→G modification of nucleotide 1078; A→C modification of nucleotide 1081; T→C modification of nucleotide 1084; T→G modification of nucleotide 1087; A→G modification of nucleotide 1090; T→C modification of nucleotide 1094; T→C modification of nucleotide 1099; T→C modification of nucleotide 1102; G→C modification of nucleotide 1105; A→C modification of nucleotide 1108; T→G modification of nucleotide 1117; A→G modification of nucleotide 1120; T→C modification of nucleotide 1123; T→C modification of nucleotide 1124; A→G modification of nucleotide 1126; T→C modification of nucleotide 1129; T→C modification of nucleotide 1132; A→G modification of nucleotide 1141; A→G modification of nucleotide 1144; A→T modification of nucleotide 1145; G→C modification of nucleotide 1146; C→G modification of nucleotide 1153; A→C modification of nucleotide 1159; T→C modification of nucleotide 1165; A→T modification of nucleotide 1166; G→C modification of nucleotide 1167; T→C modification of nucleotide 1168; T→C modification of nucleotide 1171; A→C modification of nucleotide 1174; A→G modification of nucleotide 1180; T→C modification of nucleotide 1183; T→C modification of nucleotide 1184; A→G modification of nucleotide 1186; A→G modification of nucleotide 1189; T→C modification of nucleotide 1192; A→C modification of nucleotide 1195; A→C modification of nucleotide 1199; G→C modification of nucleotide 1201; T→C modification of nucleotide 1207; T→C modification of nucleotide 1210; A→G modification of nucleotide 1216; A→G modification of nucleotide 1219; A→G modification of nucleotide 1222; A→C modification of nucleotide 1234; A→T modification of nucleotide 1235; G→C modification of nucleotide 1236; A→C modification of nucleotide 1238; A→C modification of nucleotide 1240; A→G modification of nucleotide 1255; T→C modification of nucleotide 1258; C→G modification of nucleotide 1261; T→C modification of nucleotide 1267; A→T modification of nucleotide 1268; G→C modification of nucleotide 1269; T→C modification of nucleotide 1270; T→C modification of nucleotide 1273; G→C modification of nucleotide 1279; T→C modification of nucleotide 1282; T→C modification of nucleotide 1285; A→G modification of nucleotide 1288; A→G modification of nucleotide 1291; T→C modification of nucleotide 1294; T→C modification of nucleotide 1297; A→C modification of nucleotide 1300; A→G modification of nucleotide 1306; T→C modification of nucleotide 1309; T→C modification of nucleotide 1310; A→G modification of nucleotide 1312; A→C modification of nucleotide 1315; T→G modification of nucleotide 1318; A→C modification of nucleotide 1327; T→C modification of nucleotide 1330; A→C modification of nucleotide 1333; A→T modification of nucleotide 1334; G→C modification of nucleotide 1335; T→C modification of nucleotide 1336; A→G modification of nucleotide 1339; T→C modification of nucleotide 1342; T→G modification of nucleotide 1345; T→G modification of nucleotide 1348; T→C modification of nucleotide 1357; T→G modification of nucleotide 1360; T→C modification of nucleotide 1366; C→G modification of nucleotide 1369; A→C modification of nucleotide 1370; G→C modification of nucleotide 1372; T→C modification of nucleotide 1375; T→G modification of nucleotide 1378; A→C modification of nucleotide 1381; T→C modification of nucleotide 1384; T→C modification of nucleotide 1387; A→C modification of nucleotide 1390; T→C modification of nucleotide 1393; A→G modification of nucleotide 1396; A→G modification of nucleotide 1402; G→C modification of nucleotide 1405; A→G modification of nucleotide 1411; T→C modification of nucleotide 1414; C→G modification of nucleotide 1417; T→G modification of nucleotide 1420; G→C modification of nucleotide 1423; T→C modification of nucleotide 1426; T→G modification of nucleotide 1429; T→G modification of nucleotide 1432; T→G modification of nucleotide 1438; T→C modification of nucleotide 1444; A→C modification of nucleotide 1447; T→C modification of nucleotide 1450; T→C modification of nucleotide 1453; A→C modification of nucleotide 1456; T→C modification of nucleotide 1459; C→G modification of nucleotide 1465; T→C modification of nucleotide 1471; A→C modification of nucleotide 1474; T→C modification of nucleotide 1477; A→C modification of nucleotide 1480; T→C modification of nucleotide 1483; T→G modification of nucleotide 1486; T→C modification of nucleotide 1489; C→G modification of nucleotide 1495; T→C modification of nucleotide 1498; T→C modification of nucleotide 1504; T→C modification of nucleotide 1507; T→C modification of nucleotide 1510; C→G modification of nucleotide 1528; A→G modification of nucleotide 1534; T→C modification of nucleotide 1540; T→C modification of nucleotide 1543; A→C modification of nucleotide 1546; T→C modification of nucleotide 1552; T→G modification of nucleotide 1555; T→C modification of nucleotide 1561; T→C modification of nucleotide 1564; C→G modification of nucleotide 1567; T→G modification of nucleotide 1570; G→C modification of nucleotide 1573; T→C modification of nucleotide 1576; T→C modification of nucleotide 1579; A→C modification of nucleotide 1583; G→C modification of nucleotide 1585; A→G modification of nucleotide 1588; T→C modification of nucleotide 1597; T→C modification of nucleotide 1603; T→C modification of nucleotide 1612; T→C modification of nucleotide 1615; T→G modification of nucleotide 1618; T→C modification of nucleotide 1621; T→C modification of nucleotide 1624; A→T modification of nucleotide 1631; G→C modification of nucleotide 1632; A→C modification of nucleotide 1636; A→G modification of nucleotide 1642; A→G modification of nucleotide 1645; A→C modification of nucleotide 1648; T→C modification of nucleotide 1663; A→C modification of nucleotide 1666; T→G modification of nucleotide 1669; T→G modification of nucleotide 1675; T→C modification of nucleotide 1681; A→C modification of nucleotide 1684; A→G modification of nucleotide 1687; A→C modification of nucleotide 1690; T→G modification of nucleotide 1693; T→G modification of nucleotide 1699; C→G modification of nucleotide 1705; T→C modification of nucleotide 1706; A→G modification of nucleotide 1708; A→C modification of nucleotide 1714; A→C modification of nucleotide 1717; T→C modification of nucleotide 1729; A→C modification of nucleotide 1735; T→C modification of nucleotide 1741; T→C modification of nucleotide 1744; T→G modification of nucleotide 1747; A→C modification of nucleotide 1750; T→C modification of nucleotide 1751; T→C modification of nucleotide 1756; A→C modification of nucleotide 1759; T→C modification of nucleotide 1762; T→G modification of nucleotide 1765; T→G modification of nucleotide 1771; T→C modification of nucleotide 1774; A→C modification of nucleotide 1780; T→C modification of nucleotide 1783; T→C modification of nucleotide 1786; A→C modification of nucleotide 1789; T→C modification of nucleotide 1792; T→C modification of nucleotide 1793; T→C modification of nucleotide 1798; T→C modification of nucleotide 1807; T→C modification of nucleotide 1813; A→C modification of nucleotide 1816; A→C modification of nucleotide 1819; A→G modification of nucleotide 1822; T→G modification of nucleotide 1825; T→C modification of nucleotide 1829; T→G modification of nucleotide 1834; T→G modification of nucleotide 1837; T→C modification of nucleotide 1840; A→C modification of nucleotide 1843; T→C modification of nucleotide 1846; T→C modification of nucleotide 1852; T→C modification of nucleotide 1864; G→C modification of nucleotide 1867; T→C modification of nucleotide 1876; G→C modification of nucleotide 1879; A→C modification of nucleotide 1882; A→G modification of nucleotide 1891; T→C modification of nucleotide 1894; G→C modification of nucleotide 1903; C→G modification of nucleotide 1906; A→G modification of nucleotide 1915; A→G modification of nucleotide 1918; A→C modification of nucleotide 1921; T→G modification of nucleotide 1924; A→G modification of nucleotide 1927; T→C modification of nucleotide 1930; T→C modification of nucleotide 1933; A→C modification of nucleotide 1939; T→G modification of nucleotide 1945; A→G modification of nucleotide 1954; T→C modification of nucleotide 1957; G→C modification of nucleotide 1960; T→C modification of nucleotide 1963; T→G modification of nucleotide 1966; A→C modification of nucleotide 1975; T→C modification of nucleotide 1978; T→G modification of nucleotide 1981; A→C modification of nucleotide 1985; A→C modification of nucleotide 1987; A→C modification of nucleotide 1990; A→G modification of nucleotide 1993; T→C modification of nucleotide 1996; T→C modification of nucleotide 2002; A→C modification of nucleotide 2005; T→C modification of nucleotide 2006; T→C modification of nucleotide 2011; T→G modification of nucleotide 2023; T→C modification of nucleotide 2026; T→C modification of nucleotide 2029; T→C modification of nucleotide 2035; A→G modification of nucleotide 2038; A→G modification of nucleotide 2047; A→C modification of nucleotide 2048; A→C modification of nucleotide 2050; A→G modification of nucleotide 2053; T→C modification of nucleotide 2056; A→C modification of nucleotide 2059; G→C modification of nucleotide 2062; C→G modification of nucleotide 2065; C→G modification of nucleotide 2068; T→C modification of nucleotide 2071; A→G modification of nucleotide 2074; T→G modification of nucleotide 2077; T→C modification of nucleotide 2080; A→G modification of nucleotide 2092; T→C modification of nucleotide 2095; A→C modification of nucleotide 2098; A→G modification of nucleotide 2101; T→C modification of nucleotide 2104; T→C modification of nucleotide 2107; A→G modification of nucleotide 2110; T→C modification of nucleotide 2113; A→C modification of nucleotide 2119; A→C modification of nucleotide 2125; T→C modification of nucleotide 2128; T→C modification of nucleotide 2131; A→C modification of nucleotide 2137; T→C modification of nucleotide 2141; A→G modification of nucleotide 2143; T→G modification of nucleotide 2146; T→C modification of nucleotide 2152; A→C modification of nucleotide 2153; G→C modification of nucleotide 2155; T→C modification of nucleotide 2158; A→G modification of nucleotide 2161; T→C modification of nucleotide 2164; G→C modification of nucleotide 2170; A→C modification of nucleotide 2173; T→C modification of nucleotide 2176; A→C modification of nucleotide 2179; T→C modification of nucleotide 2182; T→C modification of nucleotide 2185; T→G modification of nucleotide 2188; T→C modification of nucleotide 2189; A→C modification of nucleotide 2197; T→C modification of nucleotide 2203; A→C modification of nucleotide 2215; A→G modification of nucleotide 2218; T→C modification of nucleotide 2221; A→G modification of nucleotide 2230; G→C modification of nucleotide 2233; T→G modification of nucleotide 2236; A→C modification of nucleotide 2239; T→G modification of nucleotide 2242; A→T modification of nucleotide 2252; G→C modification of nucleotide 2253; T→C modification of nucleotide 2257; C→G modification of nucleotide 2260; A→C modification of nucleotide 2264; A→C modification of nucleotide 2266; A→C modification of nucleotide 2269; A→C modification of nucleotide 2275; T→C modification of nucleotide 2281; T→C modification of nucleotide 2287; T→C modification of nucleotide 2293; T→C modification of nucleotide 2296; T→C modification of nucleotide 2299; T→G modification of nucleotide 2302; A→C modification of nucleotide 2305; T→G modification of nucleotide 2308; T→C modification of nucleotide 2311; T→C modification of nucleotide 2314; T→C modification of nucleotide 2317; A→C modification of nucleotide 2326; A→G modification of nucleotide 2329; T→G modification of nucleotide 2335; A→C modification of nucleotide 2338; T→C modification of nucleotide 2341; T→C modification of nucleotide 2350; T→C modification of nucleotide 2353; T→C modification of nucleotide 2356; A→T modification of nucleotide 2357; G→C modification of nucleotide 2358; T→C modification of nucleotide 2362; T→C modification of nucleotide 2365; T→C modification of nucleotide 2368; T→C modification of nucleotide 2371; T→C modification of nucleotide 2377; T→C modification of nucleotide 2380; A→C modification of nucleotide 2383; T→C modification of nucleotide 2386; T→C modification of nucleotide 2395; T→C modification of nucleotide 2399; T→C modification of nucleotide 2404; A→G modification of nucleotide 2422; T→C modification of nucleotide 2425; T→C modification of nucleotide 2434; T→C modification of nucleotide 2447; T→C modification of nucleotide 2449; T→C modification of nucleotide 2452; T→C modification of nucleotide 2458; A→C modification of nucleotide 2461; T→G modification of nucleotide 2464; T→C modification of nucleotide 2467; G→C modification of nucleotide 2470; A→C modification of nucleotide 2476; T→C modification of nucleotide 2479; G→C modification of nucleotide 2485; T→C modification of nucleotide 2488; C→G modification of nucleotide 2500; T→C modification of nucleotide 2506; G→C modification of nucleotide 2509; T→C modification of nucleotide 2515; A→C modification of nucleotide 2518; T→C modification of nucleotide 2521; A→T modification of nucleotide 2525; G→C modification of nucleotide 2526; T→C modification of nucleotide 2530; T→C modification of nucleotide 2545; T→C modification of nucleotide 2548; T→C modification of nucleotide 2551; and deletion of nucleotides 2558-3197; k) a sequence encoding yes-associated protein 1 (YAP) and comprising one or more of the following modifications relative to SEQ ID NO: 22 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-402; T→C modification of nucleotide 408; G→C modification of nucleotide 414; G→C modification of nucleotide 423; G→C modification of nucleotide 426; T→C modification of nucleotide 429; A→G modification of nucleotide 432; G→C modification of nucleotide 435; A→G modification of nucleotide 450; G→C modification of nucleotide 453; G→C modification of nucleotide 459; T→C modification of nucleotide 462; G→C modification of nucleotide 465; G→C modification of nucleotide 474; G→C modification of nucleotide 480; G→C modification of nucleotide 489; G→C modification of nucleotide 492; A→C modification of nucleotide 498; G→C modification of nucleotide 504; A→G modification of nucleotide 507; G→C modification of nucleotide 510; A→C modification of nucleotide 513; G→C modification of nucleotide 519; G→C modification of nucleotide 522; G→C modification of nucleotide 531; G→C modification of nucleotide 534; G→C modification of nucleotide 537; A→C modification of nucleotide 543; G→C modification of nucleotide 555; T→C modification of nucleotide 558; C→G modification of nucleotide 573; G→C modification of nucleotide 579; G→C modification of nucleotide 585; G→C modification of nucleotide 603; C→G modification of nucleotide 606; C→G modification of nucleotide 618; G→C modification of nucleotide 633; A→C modification of nucleotide 661; G→C modification of nucleotide 663; C→G modification of nucleotide 666; G→C modification of nucleotide 669; G→C modification of nucleotide 696; G→C modification of nucleotide 699; A→G modification of nucleotide 708; A→C modification of nucleotide 720; A→T modification of nucleotide 727; G→C modification of nucleotide 728; T→C modification of nucleotide 729; T→C modification of nucleotide 732; T→C modification of nucleotide 735; A→C modification of nucleotide 738; T→C modification of nucleotide 744; A→C modification of nucleotide 747; A→C modification of nucleotide 750; T→C modification of nucleotide 759; A→C modification of nucleotide 762; T→C modification of nucleotide 768; T→G modification of nucleotide 771; A→C modification of nucleotide 774; T→C modification of nucleotide 777; T→C modification of nucleotide 780; T→G modification of nucleotide 781; T→C modification of nucleotide 786; A→C modification of nucleotide 789; T→C modification of nucleotide 792; T→C modification of nucleotide 795; T→C modification of nucleotide 802; A→C modification of nucleotide 807; T→C modification of nucleotide 810; T→G modification of nucleotide 813; T→C modification of nucleotide 816; T→C modification of nucleotide 819; T→C modification of nucleotide 822; A→C modification of nucleotide 825; T→C modification of nucleotide 837; A→C modification of nucleotide 840; A→G modification of nucleotide 843; C→G modification of nucleotide 846; T→C modification of nucleotide 849; A→C modification of nucleotide 855; A→C modification of nucleotide 858; T→C modification of nucleotide 861; A→C modification of nucleotide 864; A→C modification of nucleotide 870; T→C modification of nucleotide 873; T→C modification of nucleotide 879; T→G modification of nucleotide 882; A→C modification of nucleotide 885; T→C modification of nucleotide 891; T→C modification of nucleotide 894; T→C modification of nucleotide 897; A→C modification of nucleotide 903; T→C modification of nucleotide 906; T→C modification of nucleotide 909; T→C modification of nucleotide 912; A→G modification of nucleotide 915; T→C modification of nucleotide 918; A→C modification of nucleotide 924; A→C modification of nucleotide 927; T→C modification of nucleotide 930; A→C modification of nucleotide 942; A→C modification of nucleotide 948; T→C modification of nucleotide 951; T→C modification of nucleotide 954; T→C modification of nucleotide 957; A→C modification of nucleotide 961; A→C modification of nucleotide 963; T→C modification of nucleotide 970; A→G modification of nucleotide 972; T→C modification of nucleotide 975; T→C modification of nucleotide 984; A→C modification of nucleotide 990; A→C modification of nucleotide 993; A→C modification of nucleotide 996; A→C modification of nucleotide 1009; G→C modification of nucleotide 1011; C→G modification of nucleotide 1038; A→C modification of nucleotide 1041; A→T modification of nucleotide 1051; G→C modification of nucleotide 1052; T→C modification of nucleotide 1053; A→C modification of nucleotide 1056; A→C modification of nucleotide 1059; T→C modification of nucleotide 1071; G→C modification of nucleotide 1083; T→C modification of nucleotide 1086; A→C modification of nucleotide 1089; T→C modification of nucleotide 1092; T→C modification of nucleotide 1095; T→G modification of nucleotide 1098; T→C modification of nucleotide 1101; T→C modification of nucleotide 1104; A→C modification of nucleotide 1107; A→G modification of nucleotide 1113; A→G modification of nucleotide 1116; T→C modification of nucleotide 1125; T→C modification of nucleotide 1131; A→C modification of nucleotide 1134; A→G modification of nucleotide 1137; T→C modification of nucleotide 1140; T→C modification of nucleotide 1146; A→C modification of nucleotide 1149; T→C modification of nucleotide 1155; T→C modification of nucleotide 1173; A→G modification of nucleotide 1179; A→C modification of nucleotide 1185; A→C modification of nucleotide 1186; G→C modification of nucleotide 1188; T→G modification of nucleotide 1191; T→C modification of nucleotide 1197; T→C modification of nucleotide 1200; T→C modification of nucleotide 1203; A→C modification of nucleotide 1216; A→C modification of nucleotide 1218; A→T modification of nucleotide 1222; G→C modification of nucleotide 1223; T→C modification of nucleotide 1224; A→T modification of nucleotide 1228; G→C modification of nucleotide 1229; T→C modification of nucleotide 1230; T→C modification of nucleotide 1233; A→C modification of nucleotide 1236; A→G modification of nucleotide 1242; A→C modification of nucleotide 1248; A→C modification of nucleotide 1251; T→C modification of nucleotide 1260; A→T modification of nucleotide 1267; G→C modification of nucleotide 1268; A→C modification of nucleotide 1272; A→C modification of nucleotide 1278; C→G modification of nucleotide 1284; T→C modification of nucleotide 1290; A→T modification of nucleotide 1294; G→C modification of nucleotide 1295; A→G modification of nucleotide 1314; A→C modification of nucleotide 1323; A→G modification of nucleotide 1332; A→C modification of nucleotide 1351; G→C modification of nucleotide 1353; G→C modification of nucleotide 1359; A→G modification of nucleotide 1365; A→G modification of nucleotide 1371; A→G modification of nucleotide 1374; T→G modification of nucleotide 1380; G→C modification of nucleotide 1383; T→C modification of nucleotide 1390; A→G modification of nucleotide 1392; T→C modification of nucleotide 1401; A→T modification of nucleotide 1402; G→C modification of nucleotide 1403; T→C modification of nucleotide 1408; A→G modification of nucleotide 1410; A→C modification of nucleotide 1413; A→C modification of nucleotide 1416; T→C modification of nucleotide 1428; T→C modification of nucleotide 1431; G→C modification of nucleotide 1434; T→C modification of nucleotide 1437; A→G modification of nucleotide 1440; T→C modification of nucleotide 1443; A→C modification of nucleotide 1446; T→C modification of nucleotide 1452; T→C modification of nucleotide 1455; G→C modification of nucleotide 1461; T→C modification of nucleotide 1467; A→G modification of nucleotide 1473; T→C modification of nucleotide 1474; A→C modification of nucleotide 1477; A→C modification of nucleotide 1479; A→C modification of nucleotide 1482; G→C modification of nucleotide 1488; T→C modification of nucleotide 1494; A→T modification of nucleotide 1495; G→C modification of nucleotide 1496; A→C modification of nucleotide 1500; T→C modification of nucleotide 1503; T→C modification of nucleotide 1506; T→G modification of nucleotide 1512; A→T modification of nucleotide 1516; G→C modification of nucleotide 1517; T→C modification of nucleotide 1518; T→C modification of nucleotide 1527; T→C modification of nucleotide 1533; A→C modification of nucleotide 1536; T→C modification of nucleotide 1539; A→T modification of nucleotide 1543; G→C modification of nucleotide 1544; T→C modification of nucleotide 1545; A→C modification of nucleotide 1548; A→T modification of nucleotide 1552; G→C modification of nucleotide 1553; T→C modification of nucleotide 1554; A→C modification of nucleotide 1557; A→G modification of nucleotide 1560; A→T modification of nucleotide 1561; G→C modification of nucleotide 1562; A→T modification of nucleotide 1567; G→C modification of nucleotide 1568; A→T modification of nucleotide 1570; G→C modification of nucleotide 1571; A→T modification of nucleotide 1576; G→C modification of nucleotide 1577; T→C modification of nucleotide 1578; C→G modification of nucleotide 1581; T→C modification of nucleotide 1584; A→C modification of nucleotide 1587; A→C modification of nucleotide 1593; T→C modification of nucleotide 1596; A→T modification of nucleotide 1609; G→C modification of nucleotide 1610; T→C modification of nucleotide 1611; T→C modification of nucleotide 1617; T→C modification of nucleotide 1626; A→C modification of nucleotide 1629; T→C modification of nucleotide 1632; T→C modification of nucleotide 1635; T→C modification of nucleotide 1638; A→G modification of nucleotide 1647; A→T modification of nucleotide 1648; G→C modification of nucleotide 1649; A→C modification of nucleotide 1662; T→C modification of nucleotide 1674; A→C modification of nucleotide 1680; T→G modification of nucleotide 1689; A→G modification of nucleotide 1692; T→C modification of nucleotide 1698; T→C modification of nucleotide 1701; G→C modification of nucleotide 1704; A→C modification of nucleotide 1707; T→C modification of nucleotide 1710; T→G modification of nucleotide 1719; A→C modification of nucleotide 1722; A→C modification of nucleotide 1725; A→G modification of nucleotide 1731; A→C modification of nucleotide 1734; T→C modification of nucleotide 1737; A→C modification of nucleotide 1740; A→C modification of nucleotide 1749; A→G modification of nucleotide 1752; A→C modification of nucleotide 1755; A→C modification of nucleotide 1770; A→T modification of nucleotide 1771; G→C modification of nucleotide 1772; T→C modification of nucleotide 1773; A→G modification of nucleotide 1782; T→C modification of nucleotide 1785; T→C modification of nucleotide 1786; A→T modification of nucleotide 1789; G→C modification of nucleotide 1790; T→C modification of nucleotide 1791; T→C modification of nucleotide 1794; T→G modification of nucleotide 1803; T→C modification of nucleotide 1806; T→C modification of nucleotide 1818; T→G modification of nucleotide 1821; T→C modification of nucleotide 1822; T→C modification of nucleotide 1827; A→G modification of nucleotide 1839; T→C modification of nucleotide 1842; A→G modification of nucleotide 1845; A→G modification of nucleotide 1848; A→T modification of nucleotide 1849; G→C modification of nucleotide 1850; T→C modification of nucleotide 1854; T→G modification of nucleotide 1857; A→C modification of nucleotide 1860; and deletion of nucleotides 1864-5353; l) a sequence encoding wingless-type MMTV integration site family, member 2 (WNT2) and comprising one or more of the following modifications relative to SEQ ID NO: 23 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-158; C→G modification of nucleotide 167; T→C modification of nucleotide 170; C→G modification of nucleotide 173; T→C modification of nucleotide 176; A→C modification of nucleotide 179; C→G modification of nucleotide 188; C→G modification of nucleotide 194; T→C modification of nucleotide 197; C→G modification of nucleotide 203; T→C modification of nucleotide 204; C→G modification of nucleotide 215; T→C modification of nucleotide 221; C→G modification of nucleotide 227; A→T modification of nucleotide 228; G→C modification of nucleotide 229; T→C modification of nucleotide 233; A→C modification of nucleotide 236; A→C modification of nucleotide 249; A→C modification of nucleotide 251; T→C modification of nucleotide 254; A→C modification of nucleotide 257; T→C modification of nucleotide 260; A→C modification of nucleotide 270; G→C modification of nucleotide 272; T→C modification of nucleotide 281; T→C modification of nucleotide 287; A→C modification of nucleotide 293; A→T modification of nucleotide 303; G→C modification of nucleotide 304; G→C modification of nucleotide 308; T→C modification of nucleotide 314; A→C modification of nucleotide 329; A→C modification of nucleotide 335; T→C modification of nucleotide 338; T→C modification of nucleotide 347; T→C modification of nucleotide 353; T→C modification of nucleotide 362; T→C modification of nucleotide 368; T→C modification of nucleotide 377; A→C modification of nucleotide 380; A→G modification of nucleotide 389; T→C modification of nucleotide 407; A→C modification of nucleotide 432; A→C modification of nucleotide 434; T→C modification of nucleotide 437; A→T modification of nucleotide 441; G→C modification of nucleotide 442; C→G modification of nucleotide 446; T→C modification of nucleotide 449; G→C modification of nucleotide 455; T→C modification of nucleotide 458; C→G modification of nucleotide 461; C→G modification of nucleotide 464; A→C modification of nucleotide 467; A→T modification of nucleotide 468; G→C modification of nucleotide 469; T→C modification of nucleotide 470; A→T modification of nucleotide 471; G→C modification of nucleotide 472; T→C modification of nucleotide 473; G→C modification of nucleotide 476; A→G modification of nucleotide 479; G→C modification of nucleotide 482; T→C modification of nucleotide 488; T→G modification of nucleotide 491; T→C modification of nucleotide 503; A→C modification of nucleotide 506; T→C modification of nucleotide 509; T→G modification of nucleotide 515; A→G modification of nucleotide 518; T→C modification of nucleotide 521; A→C modification of nucleotide 531; G→C modification of nucleotide 533; T→C modification of nucleotide 539; A→T modification of nucleotide 540; G→C modification of nucleotide 541; A→G modification of nucleotide 545; A→C modification of nucleotide 548; A→G modification of nucleotide 551; T→C modification of nucleotide 552; A→G modification of nucleotide 554; T→C modification of nucleotide 569; T→C modification of nucleotide 572; A→C modification of nucleotide 575; A→G modification of nucleotide 584; A→C modification of nucleotide 587; A→T modification of nucleotide 588; G→C modification of nucleotide 589; T→C modification of nucleotide 590; A→T modification of nucleotide 600; G→C modification of nucleotide 601; A→G modification of nucleotide 605; T→C modification of nucleotide 623; A→T modification of nucleotide 630; G→C modification of nucleotide 631; T→C modification of nucleotide 632; T→C modification of nucleotide 638; T→C modification of nucleotide 641; G→C modification of nucleotide 650; T→C modification of nucleotide 659; T→C modification of nucleotide 665; T→C modification of nucleotide 671; A→G modification of nucleotide 674; T→C modification of nucleotide 677; A→C modification of nucleotide 687; G→C modification of nucleotide 689; A→G modification of nucleotide 692; T→C modification of nucleotide 701; A→C modification of nucleotide 705; A→C modification of nucleotide 707; T→G modification of nucleotide 722; A→C modification of nucleotide 732; A→C modification of nucleotide 734; T→C modification of nucleotide 737; A→C modification of nucleotide 740; A→C modification of nucleotide 741; G→C modification of nucleotide 743; T→C modification of nucleotide 749; A→G modification of nucleotide 752; T→C modification of nucleotide 762; A→G modification of nucleotide 767; A→G modification of nucleotide 770; A→G modification of nucleotide 773; T→C modification of nucleotide 782; T→C modification of nucleotide 785; T→C modification of nucleotide 788; A→T modification of nucleotide 792; G→C modification of nucleotide 793; T→C modification of nucleotide 794; T→C modification of nucleotide 803; T→C modification of nucleotide 806; A→C modification of nucleotide 810; G→C modification of nucleotide 812; A→C modification of nucleotide 815; T→C modification of nucleotide 833; A→C modification of nucleotide 840; G→C modification of nucleotide 842; A→G modification of nucleotide 845; A→C modification of nucleotide 848; T→C modification of nucleotide 857; C→G modification of nucleotide 860; A→C modification of nucleotide 864; G→C modification of nucleotide 866; T→C modification of nucleotide 875; G→C modification of nucleotide 878; A→G modification of nucleotide 890; C→G modification of nucleotide 893; T→C modification of nucleotide 905; T→C modification of nucleotide 911; T→C modification of nucleotide 920; A→G modification of nucleotide 923; T→C modification of nucleotide 929; A→C modification of nucleotide 933; G→C modification of nucleotide 935; T→C modification of nucleotide 938; A→C modification of nucleotide 947; G→C modification of nucleotide 950; A→G modification of nucleotide 953; T→C modification of nucleotide 956; C→G modification of nucleotide 962; T→C modification of nucleotide 968; T→C modification of nucleotide 971; T→C modification of nucleotide 977; T→C modification of nucleotide 980; A→C modification of nucleotide 983; T→C modification of nucleotide 992; A→C modification of nucleotide 996; G→C modification of nucleotide 998; A→C modification of nucleotide 1004; A→C modification of nucleotide 1010; T→C modification of nucleotide 1022; A→C modification of nucleotide 1025; G→C modification of nucleotide 1028; T→C modification of nucleotide 1034; T→C modification of nucleotide 1044; T→C modification of nucleotide 1049; A→C modification of nucleotide 1055; A→T modification of nucleotide 1065; G→C modification of nucleotide 1066; A→G modification of nucleotide 1073; T→G modification of nucleotide 1076; T→C modification of nucleotide 1082; T→C modification of nucleotide 1085; G→C modification of nucleotide 1088; A→C modification of nucleotide 1089; A→C modification of nucleotide 1091; T→C modification of nucleotide 1097; A→C modification of nucleotide 1103; C→G modification of nucleotide 1112; G→C modification of nucleotide 1118; T→C modification of nucleotide 1130; T→C modification of nucleotide 1136; A→G modification of nucleotide 1139; T→C modification of nucleotide 1154; T→C modification of nucleotide 1166; A→C modification of nucleotide 1199; A→T modification of nucleotide 1215; G→C modification of nucleotide 1216; T→C modification of nucleotide 1217; and deletion of nucleotides 1229-2115; and/or m) a sequence encoding wingless-type MMTV integration site family, member 9B (WNT9b) and comprising one or more of the following modifications relative to SEQ ID NO: 24 (e.g., one, two, three, four, or all of the following modifications): deletion of nucleotides 1-52; G→C modification of nucleotide 64; G→C modification of nucleotide 70; T→C modification of nucleotide 82; G→C modification of nucleotide 85; G→C modification of nucleotide 88; T→C modification of nucleotide 106; T→C modification of nucleotide 112; T→C modification of nucleotide 145; T→C modification of nucleotide 148; C→G modification of nucleotide 154; A→C modification of nucleotide 160; A→C modification of nucleotide 169; T→C modification of nucleotide 178; G→C modification of nucleotide 181; A→C modification of nucleotide 184; A→C modification of nucleotide 187; G→C modification of nucleotide 193; A→C modification of nucleotide 196; T→C modification of nucleotide 202; T→C modification of nucleotide 205; T→C modification of nucleotide 208; T→C modification of nucleotide 223; A→G modification of nucleotide 229; A→C modification of nucleotide 242; G→C modification of nucleotide 244; G→C modification of nucleotide 247; C→G modification of nucleotide 259; A→C modification of nucleotide 263; G→C modification of nucleotide 265; G→C modification of nucleotide 268; T→C modification of nucleotide 283; A→C modification of nucleotide 293; G→C modification of nucleotide 295; T→C modification of nucleotide 298; T→C modification of nucleotide 301; A→C modification of nucleotide 304; G→C modification of nucleotide 313; A→G modification of nucleotide 322; T→C modification of nucleotide 325; A→C modification of nucleotide 338; G→C modification of nucleotide 340; A→T modification of nucleotide 359; G→C modification of nucleotide 360; G→C modification of nucleotide 370; A→C modification of nucleotide 371; G→C modification of nucleotide 373; T→C modification of nucleotide 376; C→G modification of nucleotide 385; A→C modification of nucleotide 389; A→C modification of nucleotide 391; T→C modification of nucleotide 397; G→C modification of nucleotide 406; T→C modification of nucleotide 418; A→C modification of nucleotide 421; T→C modification of nucleotide 427; A→C modification of nucleotide 430; T→C modification of nucleotide 433; C→G modification of nucleotide 439; G→C modification of nucleotide 442; T→C modification of nucleotide 445; A→C modification of nucleotide 448; T→C modification of nucleotide 455; G→C modification of nucleotide 457; A→T modification of nucleotide 464; G→C modification of nucleotide 465; T→C modification of nucleotide 466; T→C modification of nucleotide 469; G→C modification of nucleotide 472; T→C modification of nucleotide 487; T→C modification of nucleotide 490; T→C modification of nucleotide 493; A→C modification of nucleotide 505; A→T modification of nucleotide 515; G→C modification of nucleotide 516; G→C modification of nucleotide 520; T→C modification of nucleotide 538; T→C modification of nucleotide 544; T→C modification of nucleotide 547; T→C modification of nucleotide 553; A→T modification of nucleotide 563; G→C modification of nucleotide 564; C→G modification of nucleotide 577; A→T modification of nucleotide 578; G→C modification of nucleotide 579; G→C modification of nucleotide 592; A→C modification of nucleotide 599; A→C modification of nucleotide 601; A→C modification of nucleotide 604; A→T modification of nucleotide 605; G→C modification of nucleotide 606; A→C modification of nucleotide 617; G→C modification of nucleotide 619; G→C modification of nucleotide 622; A→C modification of nucleotide 623; G→C modification of nucleotide 625; T→C modification of nucleotide 628; T→C modification of nucleotide 661; A→T modification of nucleotide 668; G→C modification of nucleotide 669; A→C modification of nucleotide 677; A→C modification of nucleotide 679; A→C modification of nucleotide 682; T→C modification of nucleotide 697; T→C modification of nucleotide 700; A→C modification of nucleotide 706; T→C modification of nucleotide 715; T→C modification of nucleotide 718; T→G modification of nucleotide 721; T→C modification of nucleotide 724; T→C modification of nucleotide 730; C→G modification of nucleotide 742; G→C modification of nucleotide 748; T→C modification of nucleotide 751; A→G modification of nucleotide 778; T→C modification of nucleotide 784; G→C modification of nucleotide 790; T→C modification of nucleotide 793; C→G modification of nucleotide 796; A→T modification of nucleotide 806; G→C modification of nucleotide 807; T→C modification of nucleotide 808; T→C modification of nucleotide 824; T→C modification of nucleotide 829; T→C modification of nucleotide 832; A→G modification of nucleotide 841; T→C modification of nucleotide 853; A→C modification of nucleotide 859; T→C modification of nucleotide 862; T→C modification of nucleotide 865; A→G modification of nucleotide 880; T→C modification of nucleotide 886; T→C modification of nucleotide 889; G→C modification of nucleotide 895; C→G modification of nucleotide 904; A→G modification of nucleotide 913; T→C modification of nucleotide 916; T→C modification of nucleotide 919; A→T modification of nucleotide 923; G→C modification of nucleotide 924; G→C modification of nucleotide 934; A→T modification of nucleotide 938; G→C modification of nucleotide 939; T→C modification of nucleotide 949; G→C modification of nucleotide 952; G→C modification of nucleotide 958; A→C modification of nucleotide 961; A→C modification of nucleotide 965; G→C modification of nucleotide 967; T→C modification of nucleotide 973; T→C modification of nucleotide 976; A→C modification of nucleotide 979; A→T modification of nucleotide 986; G→C modification of nucleotide 987; T→C modification of nucleotide 988; A→T modification of nucleotide 992; G→C modification of nucleotide 993; A→T modification of nucleotide 995; G→C modification of nucleotide 996; A→G modification of nucleotide 1000; T→C modification of nucleotide 1006; G→C modification of nucleotide 1009; A→C modification of nucleotide 1012; A→T modification of nucleotide 1028; G→C modification of nucleotide 1029; T→G modification of nucleotide 1042; T→C modification of nucleotide 1057; T→C modification of nucleotide 1096; A→C modification of nucleotide 1099; C→G modification of nucleotide 1111; T→C modification of nucleotide 1117; and deletion of nucleotides 1130-4519.

›SUMMARY · 2 of 3

In some embodiments of any of the aspects, the mRNA comprises one of the foregoing sequences comprising each of the listed modifications.

In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA comprises one or more of:

a) SEQ ID NO: 8 (GH); b) SEQ ID NO: 9 (EGF); c) SEQ ID NO: 10 (HGF); d) SEQ ID NO: 11 (p21); e) SEQ ID NO: 12 (VEGF165); f) SEQ ID NO: 13 (IGF-1); g) SEQ ID NO: 14 (IGF-1 IL-2 SP); h) SEQ ID NO: 15 (secreted EGF); i) SEQ ID NO: 16 (stat5bca); j) SEQ ID NO: 21 (beta catenin); k) SEQ ID NO: 25 (YAP); l) SEQ ID NO: 26 (WNT2); and m) SEQ ID NO: 27 (WNT9B).

In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA further comprises at least one modified nucleoside.

In one aspect of any of the embodiments, described herein is a composition comprising at least one liver regenerative factor mRNAs comprising at least one modified nucleoside; wherein the at least one liver regenerative factor is selected from the group consisting of: vascular endothelial growth factor A (VEGFA); hepatocyte growth factor (HGF); growth hormone (GH); insulin-like growth factor 1 (IGF-1), epidermal growth factor (EGF); signal transducer and activator of transcription 5B (STAT5b); cyclin-dependent kinase inhibitor 1A (p21); beta catenin (CTNNB1); yes-associated protein (YAP); wingless-type MMTV integration site family, member 2 (WNT2); and wingless-type MMTV integration site family, member 9B (WNT9b).

In some embodiments of any of the aspects, the composition further comprises a carrier complexed with the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside. In some embodiments of any of the aspects, the carrier is a nanoparticle. In some embodiments of any of the aspects, the carrier is a polymer nanoparticle. In some embodiments of any of the aspects, the nanoparticle is a lipid nanoparticle (LNP).

In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of: VEGFA; HGF; GH; and EGF. In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of: HGF; GH; EGF; and p21. In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of: HGF; GH; and EGF. In some embodiments of any of the aspects, the at least one liver regenerative factor comprises two or more liver regenerative factors selected from the group consisting of: VEGFA; HGF; GH; IGF-1; EGF; STAT5bCA; p21; CNNTB1; YAP; WNT2; and WNT9b.

In some embodiments of any of the aspects, the at least one liver regenerative factor is a human liver regenerative factor or a murine liver regenerative factor.

In some embodiments of any of the aspects, the at least one modified nucleoside comprises at least one non-natural nucleoside. In some embodiments of any of the aspects, the at least one modified nucleoside is selected from the group consisting of: pseudouridine, N1-methylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine, pseudouridine, and mixtures thereof, 5-methyluridine, 5-methoxyuridine, and 2′-O-methyluridine. In some embodiments of any of the aspects, the at least one modified nucleoside comprises at least one-methylpseudouridine (m1Ψ)-5′ triphosphate (TriLink).

In some embodiments of any of the aspects, the LNP comprises at least one ionizable lipid, at least one phospholipid, at least one structured lipid, and at least one polyethylene glycol (PEG)-lipid. In some embodiments of any of the aspects, the at least one ionizable lipid is selected from the group consisting of: 2,2-dioleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA); dioleyl-methyl 4-dimethylaminobutyrate (DLin-MC3-DMA); and di ((Z)-non-2-en-1-yl) 9-((4-(dimethylamino) butyryl) oxy) heptadecanedioate (L319). In some embodiments of any of the aspects, the at least one ionizable lipid has a pKA in the range of 6.0-6.5. In some embodiments of any of the aspects, the at least one phospholipid comprises phosphatidylcholine. In some embodiments of any of the aspects, the at least one structured lipid comprises cholesterol.

In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is encapsulated in the nanoparticle. In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is in an aqueous solution and in admixture with an ethanolic lipid mixture at acidic pH. In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is complexed to the LNP in a way selected from the group consisting of: encapsulation in the interior of the LNP; interspersed within the lipid bilayer of the LNP; and attached to the LNP via a linking molecule.

In some embodiments of any of the aspects, the liver regenerative factor comprises GH; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 8. In some embodiments of any of the aspects, the liver regenerative factor comprises EGF; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 9. In some embodiments of any of the aspects, the liver regenerative factor comprises HGF; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 10. In some embodiments of any of the aspects, the liver regenerative factor comprises p21; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 11. In some embodiments of any of the aspects, the liver regenerative factor comprises VEGF; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 12. In some embodiments of any of the aspects, the liver regenerative factor comprises IGF-1; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 13. In some embodiments of any of the aspects, the liver regenerative factor comprises IGF-1 IL-2 SP; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 14. In some embodiments of any of the aspects, the liver regenerative factor comprises secreted EGF; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 15. In some embodiments of any of the aspects, the liver regenerative factor comprises stat5b; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 16. In some embodiments of any of the aspects, the liver regenerative factor comprises beta catenin; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 21. In some embodiments of any of the aspects, the liver regenerative factor comprises YAP; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 25. In some embodiments of any of the aspects, the liver regenerative factor comprises WNT2; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 26. In some embodiments of any of the aspects, the liver regenerative factor comprises WNT9b; and the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater) sequence identity to the nucleotide sequence set forth in SEQ ID NO: 27.

›SUMMARY · 3 of 3

In some embodiments of any of the aspects, the composition further comprises N-acetyl cysteine (NAC). In one aspect of any of the embodiments, described herein is the combination of a composition described herein and N-acetyl cysteine (NAC).

In one aspect of any of the embodiments, described herein is a method of treating liver injury or liver disease in a subject in need thereof, the method comprising administering the composition or combination as described herein to the subject. In one aspect of any of the embodiments, described herein is a method of accelerating intrinsic liver repair in a subject in need thereof, the method comprising administering the composition or combination as described herein to the subject. In some embodiments of any of the aspects, the subject is a subject in need of treatment for acute liver disease, chronic liver disease, or acetaminophen (acetyl-para-aminophenol, APAP) overdose. In some embodiments of any of the aspects, the acute or chronic liver disease is selected from the group consisting of: haemophilia; familial hypercholesterolemia; ornithine transcarbamylase deficiency; α-antitrypsin deficiency; non-alcoholic fatty liver disease (NAFLD); non-alcoholic steatohepatitis (NASH); liver fibrosis; liver cirrhosis; alcoholic fatty liver disease; alcohol-related liver disease (ARLD); phenylketonuria; glycogen storage disease; α1-antitrypsin deficiency; hereditary hemochromatosis; tyrosinemia type 1; argininosuccinic aciduria; hepatitis virus infection; non-viral hepatitis; autoimmune hepatitis; primary biliary cholangitis; cirrhosis, biliary atresia; liver cancer; genetic cholestasis; hemochromatosis; Gilbert syndrome; primary sclerosing cholangitis (PSC); and Wilson's disease. In some embodiments of any of the aspects, the method further comprises administering N-acetyl cysteine (NAC) to the subject.

In one aspect of any of the embodiments, described herein is a method of engrafting cells in a liver tissue, the method comprising introducing the cells into the liver tissue and contacting the cells or the liver tissue with the composition or combination described herein. In some embodiments of any of the aspects, the cells are primary human hepatocytes (PHH) or induced pluripotent stem cell-derived hepatocyte-like-cells (iPSC-HLCs). In some embodiments of any of the aspects, the liver tissue is a liver in a subject, the introducing comprises transplanting the cells into the liver, and the contacting comprises administering. In some embodiments of any of the aspects, the subject is a subject in need of treatment for acute liver disease, chronic liver disease, or genetic liver disease. In some embodiments of any of the aspects, the acute or chronic liver disease is selected from the group consisting of: haemophilia; familial hypercholesterolemia; ornithine transcarbamylase deficiency; α-antitrypsin deficiency; non-alcoholic fatty liver disease (NAFLD); non-alcoholic steatohepatitis (NASH); liver fibrosis; liver cirrhosis; alcoholic fatty liver disease; alcohol-related liver disease (ARLD); phenylketonuria; glycogen storage disease; α1-antitrypsin deficiency; hereditary hemochromatosis; tyrosinemia type 1; argininosuccinic aciduria; hepatitis virus infection; non-viral hepatitis; autoimmune hepatitis; primary biliary cholangitis; cirrhosis, biliary atresia; liver cancer; genetic cholestasis; hemochromatosis; Gilbert syndrome; primary sclerosing cholangitis (PSC); and Wilson's disease. In some embodiments of any of the aspects, the subject is a subject in need of treatment for alpha-1 antitrypsin deficiency associated liver disease (AATD).

In some embodiments of any of the aspects, the composition or combination is administered once. In some embodiments of any of the aspects, the composition or combination is administered twice or more. In some embodiments of any of the aspects, the composition or combination comprises two or more liver regenerative factor mRNAs. In some embodiments of any of the aspects, the method comprising administering a first composition or combination comprising a first liver regenerative factor mRNA and concurrently administering a second composition or combination comprising a second liver regenerative factor mRNA. In some embodiments of any of the aspects, the method comprises administering a first composition or combination comprising a first liver regenerative factor mRNA and separately administering a second composition or combination comprising a second liver regenerative factor mRNA.

In some embodiments of any of the aspects, the administering is intravenous administration. In some embodiments of any of the aspects, the administering is via the common bile duct or to the gallbladder.

›BRIEF DESCRIPTION OF THE DRAWINGS · 1 of 10

FIGS. 1 A -ID depict VEGFR signaling regulating BEC-driven liver regeneration in zebrafish. ( FIG. 1 A ) Single-optical section images showing the expression of Bhmt, Tp1:H2B-mCherry, and fabp10a:CFP-NTR in regenerating livers (dotted lines) at R24h. Scheme illustrates the periods of Mtz and SU5416 treatments and an analysis stage. Quantification of the percentage of hepatocytes (Bhmt + ) among BEC-derived cells (H2B-mCherry + ) and quantification of liver size are shown. ( FIG. 1 B ) Whole-mount in situ hybridization images showing gc and f5 expression (arrows) in regenerating livers at R24h. Numbers in the upper-right corner indicate the proportion of larvae exhibiting the phenotype shown. Based on the levels of hepatic gc and f5 expression, larvae were divided into three groups: no, weak, and strong. ( FIG. 1 C ) Single-optical section images showing the expression of Tp1:H2B-mCherry, fabp10a:CFP-NTR, and Bhmt in regenerating livers (dashed lines) at R26h. To overexpress sFlt1, Tg(hs:sflt1) larvae were heat-shocked four times at A13h, A24h, A35h, and R10h. Quantification of the percentage of Bhmt + among BEC-derived cells and quantification of liver size are shown. ( FIG. 1 D ) Maximum projection images showing the expression of hs:loxP-mCherry-loxP and Bhmt in regenerating livers (dashed lines) at R3h. The Tg(Tp1:CreERT2) and Tg(hs:loxP-mCherry-loxP-hVEGFA) lines were used to express hVEGFA in a subset of BEC-derived cells during regeneration. Larvae were treated with 10 μM 4-OHT from 2.5 to 3.5 dpf for 24 hours, heat-shocked twice at A20h and A34h, and harvested at R3h. Quantification of the percentage of Bhmt + area in the liver is shown. Data are presented as mean±SEM. (D) **p<0.01, ***p<0.001, and ****p<0.0001; statistical significance was calculated using an unpaired two-tailed t-test. Scale bars: 50 ( FIGS. 1 A, 1 C, 1 D ), 100 ( FIG. 1 B ) μm.

FIGS. 2 A- 2 I depict VEGFA mRNA-LNP administration inducing BEC-to-hepatocyte conversion and promoting liver repair in CDE/p21-induced chronic liver injury in mice. ( FIG. 2 A ) Scheme showing key interventions in the experimental design using the KRT19-Cre ERT , R26 LSL tdTomato mice. AAV8-Tbg-p21 vector was administered to induce hepatocyte senescence. Injury was induced by CDE diet for 2 weeks, followed by VEGFA mRNA-LNP or control Poly(C) RNA-LNP injections, 10 μg/20 g body weight. ( FIG. 2 B ) Human-specific VEGFA ELISA in mouse serum (n=3), injected with VEGFA mRNA-LNPs. Serum was collected 5 h, 24 h, 48 h, and 72 h after injection. ( FIG. 2 C ) Immunofluorescent images showing tdTomato+ clusters (outlined) in livers of mice given two injections of either control Poly(C) RNA-LNP (n=4) or VEGFA mRNA-LNP (n=4). The bar graph shows quantification of tdTomato+ areas in the two groups. ( FIG. 2 D ) Representative images of tdTomato+ and KRT7+ liver cells in Poly(C) RNA-LNP- or VEGFA mRNA-LNP-treated mice. The close-up images highlight tdTomato+ hepatocytes (arrowheads) adjacent to tdTomato+ Krt7+ BECs (arrows) in VEGFA-mRNA-LNP-treated mice. * represents tdTomato+ BEC areas. ( FIG. 2 E ) tdTomato and PAS staining on serial sections of liver tissue demonstrating glycogen storage in tdTomato+ hepatocytes. ( FIG. 2 F ) Histograms from flow cytometry of hepatocyte fraction isolated from mouse livers. Values of histograms represent % tdTomato+ population in the hepatocyte fraction and hepatocytes from a control non-tdTomato background run simultaneously with the experimental mice. Bar graph shows the total % tdTomato+ hepatocytes calculated by extrapolating the lineage tracing efficiency as 100% across all mice. ( FIG. 2 G ) Representative brightfield images and corresponding bar graph showing quantification of % trichrome stained area estimated from at least three different fields in each mouse in the two groups (n=4 per group). ( FIG. 2 H ) LipidSpot staining showing the accumulation of lipid droplets in hepatocytes. The bar graph shows quantification of % LipidSpot stained area averaged from three different fields in each mouse in the two groups (n=4 per group). ( FIG. 2 I ) Bar graph depicting total serum cholesterol levels in mice. Numerical data are presented as mean±s.d. P values were determined by two-tailed Student's t-test, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

FIGS. 3 A- 3 F depict VEGFA mRNA-LNP administration inducing BEC-to-hepatocyte conversion in APAP/p21-induced acute liver injury in mice. ( FIG. 3 A ) Experimental design using the Krt19-Cre ERT , R26 LSL tdTomato mice. AAV8-Tbg-p21 vector was administered to induce hepatocyte senescence. Injury was induced by a single intra-peritoneal injection of acetaminophen (APAP), followed by VEGFA mRNA-LNP or control Poly(C) RNA-LNP injections, 10 μg/20 g body weight. ( FIG. 3 B ) Representative immunofluorescent images showing comparison of EpCAM+ BECs observed in mice treated with APAP or APAP/p21. ( FIG. 3 C ) Immunofluorescence microscopy images of tdTomato+ areas (outlined) in livers of mice given two injections of either control Poly(C) RNA-LNP (n=3) or VEGFA mRNA-LNP (n=3). The graph shows quantification of tdTomato+ area in both groups. ( FIG. 3 D ) Histograms from flow cytometry of hepatocyte fraction isolated from mouse livers. Values of histograms represent % tdTomato+ population in the hepatocyte fraction and hepatocytes from a control non-tdTomato background run simultaneously with experimental mouse. Bar graph shows the total % tdTomato+ hepatocytes calculated by extrapolating the lineage tracing efficiency as 100% across all mice. ( FIG. 3 E ) Representative immunofluorescence microscopy images showing hepatocyte identity of tdTomato+ cells (arrows) with HNF4α staining in the VEGFA mRNA-LNP-treated group. Close-up images show HNF4α+ cells within a tdTomato+ biliary duct. ( FIG. 3 F ) tdTomato and PAS staining on serial sections of liver tissue demonstrating the ability of tdTomato+ hepatocytes to store glycogen. Data are presented as mean±s.d. P values were determined by two-tailed Student's t-test, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

›BRIEF DESCRIPTION OF THE DRAWINGS · 2 of 10

FIGS. 4 A- 4 H depict VEGFA mRNA-LNP administration inducing BEC-to-hepatocyte conversion in CDE/p21-induced chronic liver injury in Kdr-2A-Cre ERT2 -2A-eYFP mice. ( FIG. 4 A- 4 B ) Expression of KDR in BECs from the CDE- and APAP-induced liver injuries in the presence of AAV8-Tbg-p21. KDR expression was observed in some of the BECs depicted by arrowheads. ( FIG. 4 C ) Scheme showing key interventions in the experimental design using the Kdr-2A-Cre ERT2 -2A-eYFP, R26 LSL tdTomato mice to track any cell that expresses KDR using CDE/p21 injury model. Additionally, three injections of tamoxifen were given on alternate days in the second week of injury to trace any BECs expressing KDR. ( FIG. 4 D ) Representative close-up images of a bile duct showing tdTomato+ EPCAM+ BECs. ( FIG. 4 E ) Immunofluorescence microscopy images showing tdTomato+ areas (outlined) in livers of mice given 2 injections of either control Poly(C) RNA-LNP (n=3) or VEGFA mRNA-LNP (n=3). The bar graph shows the quantification of the tdTomato+ area in the two groups. ( FIG. 4 F- 4 G ) Representative immunofluorescence images depicting hepatocyte identity of tdTomato+ cells (outlined and identified with *) with HNF4α staining, a hepatocyte marker, or CD26 staining, a mature hepatocyte marker expressed on the canalicular face of the hepatocytes. ( FIG. 4 H ) Representative brightfield images of Oil Red O-stained liver tissues showing lipid accumulation in the liver. The bar graph shows the quantification of the % Oil Red O-stained area averaged from at least three different fields in each mouse in the two groups (n=3 per group). Numerical data are presented as mean±s.d. P values were determined by two-tailed Student's t-test, *p<0.05, **p<0.01.

FIGS. 5 A- 5 E depict VEGFA mRNA-LNP administration inducing BEC-to-hepatocyte conversion in APAP/p21-induced acute liver injury in Kdr-2A-Cre ERT2 -2A-eYFP mice. ( FIG. 5 A ) Scheme showing key interventions in the experimental design using the Kdr-2A-Cre ERT2 -2A-eYFP, R26 LSL tdTomato mice to track any cell that expresses KDR in APAP/p21 injury model. Three injections of tamoxifen are given on alternate days around injury to trace any BEC expressing KDR in response to APAP injury. ( FIG. 5 B ) Immunofluorescent microscopy images showing tdTomato+ areas (outlined) in livers of control firefly luciferase mRNA-LNP- (n=6) or VEGFA mRNA-LNP- (n=6) treated mice. The bar graph shows quantification of tdTomato+ area in the two groups. ( FIG. 5 C- 5 D ) Representative immunofluorescent images depicting hepatocyte identity of tdTomato+ cells (outlined and identified with *) with HNF4α staining, a hepatocyte marker or CD26 staining, a mature hepatocyte marker that is expressed on the canalicular face of the hepatocytes. ( FIG. 5 E ) Immunofluorescent images showing expression of YFP in liver cells of Kdr-2A-Cre ERT2 -2A-eYFP mice three days after acute liver toxicity by intraperitoneal APAP injection (400 mg/kg to male or 500 mg/kg to female). The fields of interest have been enlarged below. Arrowheads depict BECs co-expressing YFP and EpCAM while arrows depict large intermediate hepatocyte-like cells expressing YFP. Numerical data in the figure are presented as mean±s.d. P values were determined by two-tailed Student's t-test for comparison between two groups, ***p<0.001.

FIGS. 6 A- 6 E depict evidence of BEC-to-hepatocyte conversion and KDR expression in human liver samples from non-alcoholic steatohepatitis (NASH) cirrhosis and alcoholic cirrhosis with ESLD patients. ( FIG. 6 A ) Immunofluorescent images showing small cuboidal KRT7+ BECs (arrowheads) and KRT7+ intermediate hepatocyte-like cells (arrows) largely present in the regenerative nodules (*) close to the fibrous septa (outlined). ( FIG. 6 B ) Immunofluorescence images showing KRT7 and GS staining in human cirrhotic liver specimens. The white arrowheads depict GS+ KRT7+ BECs possessing a duct cell-like morphology. The white arrows depict the rare GS+ KRT7+ intermediate hepatocyte-like cells, while arrows depict GS+ KRT7− intermediate hepatocyte-like cells. ( FIG. 6 C ) Immunofluorescent images of cirrhotic liver specimen showing fibrous septa stained for KRT7+ BECs and KDR+ sinusoidal endothelial cells. Numerous KRT7+ KDR+ (arrowheads) BECs can be observed (magnified images) along with larger KDR+ intermediate hepatocyte-like cells (arrows) surrounding the KRT7+ KDR+ BECs. ( FIG. 6 D ) Relative gene expression of KDR, KRT7, HNF4α determined in hepatocytes (Heps) isolated from normal (n=5) or cirrhotic human livers, Child-Pugh B (n=5) and Child-Pugh C (n=4). ( FIG. 6 E ) Western blots showing protein expression of KDR, KRT7, HNF4α in hepatocytes isolated from normal or cirrhotic human livers Child-Pugh B (n=3) and Child-Pugh C (n=3) analyzed above in panel d. GAPDH serves as endogenous control. Numerical data in the figure are presented as mean±s.d. P values were determined by two-tailed Student's t-test for comparison between two groups, *p<0.05.

FIGS. 7 A- 7 G depict VEGFA mRNA-LNP administration inducing BEC-to-hepatocyte conversion and promoting liver repair in CDE/p21-induced chronic liver injury in mice ( FIG. 7 A ) Density plots from flow cytometry of non-parenchymal cell (NPC) fraction isolated from mouse livers. The NPC fraction was gated to exclude dead cell, debris, and cells expressing CD11b, CD45, Ter119, CD31. Values show % tdTomato+ EpCAM+ population. Lower panel shows cells from a control non-tdTomato mouse run simultaneously with each experimental mouse. ( FIG. 7 B ) Representative immunofluorescent microscopy images showing extent of ductular response (KRT7+ BECs) in Krt19-Cre ERT R26 LSL tdTomato mice treated with AAV8-Tbg-p21 vector and choline-deficient diet with 0.1% ethionine supplementation for 2 weeks, followed by VEGFA mRNA-LNP or control Poly(C) RNA-LNP injections, 10 μg/20 g body weight. The bar graph shows quantification of KRT7-positive area averaged from at least three different fields in each mouse in the two groups (n=4 per group). ( FIG. 7 C ) KRT7 and tdTomato staining showing decrease in BEC density in regions of BEC-to-hepatocyte conversion in VEGFA mRNA-LNP-treated group (outline). ( FIG. 7 D ) Scheme showing key interventions in the experimental design using the Krt19-Cre ERT , R26 LSL tdTomato mice without inducing hepatocyte senescence through AAV8-Tbg-p21 vector. Injury was induced by choline-deficient diet with 0.1% ethionine supplementation for 2 weeks, followed by VEGFA mRNA-LNP or control Poly(C) RNA-LNP injections, 10 μg/20 g body weight, as indicated. ( FIG. 7 E ) Representative immunofluorescent microscopy images showing extent of ductular response (KRT7+ biliary epithelial cells) in chronically injured liver in the absence of AAV8 p21-induced hepatic senescence. ( FIG. 7 F ) Close-up images of tdTomato+ liver cells in Poly(C) RNA-LNP- and VEGFA mRNA-LNP-treated mice. ( FIG. 7 G ) Representative immunofluorescent microscopy images showing tdTomato+ liver cells and p21 staining on serial sections of liver tissue demonstrating the endogenous up-regulation of p21 in areas around tdTomato+ hepatocytes. Images on the right have been magnified to reveal p21 staining. Numerical data are presented as mean±s.d. P values were determined by one-way ANOVA followed by Tukey's method for multiple comparisons. **p<0.01, ***p<0.001. ns=non-significant.

›BRIEF DESCRIPTION OF THE DRAWINGS · 3 of 10

FIGS. 8 A- 8 D depict VEGFA mRNA-LNP administration inducing BEC-to-hepatocyte conversion in APAP/p21-induced acute liver injury in mice. ( FIG. 8 A ) Representative immunofluorescence microscope images showing expression of p21-induced by AAV8-Tbg-p21 vector as compared to mice injected with a null vector. Mice were administered with 5×10 11 gc of AAV8-Tbg-p21 vector or null vector intravenously through retro-orbital sinus injection. ( FIG. 8 B ) Density plots from flow cytometry of NPC fraction isolated from mouse livers. The NPC fraction was gated to exclude dead cell, debris, and cells expressing CD11b, CD45, Ter119, CD31. Values show % tdTomato+ EpCAM+ population. Lower panel shows cells from a control non-tdTomato mouse run simultaneously with each experimental mouse. ( FIG. 8 C ) Scheme showing key interventions in the experimental design using the Krt19-Cre ERT , R26 LSL tdTomato mice without administering AAV8-Tbg-p21 vector to induce hepatocyte senescence. Injury was induced by a single intraperitoneal injection of APAP (500 mg/kg to female mice) followed by VEGFA mRNA-LNP or control Poly(C) RNA-LNP injections, 10 μg/20 g body weight. ( FIG. 8 D ) Representative close-up images of tdTomato+ liver cells in Poly(C) RNA-LNP- and VEGFA mRNA-LNP-treated mice. HNF4α staining shows the hepatocyte identity of selected tdTomato+ cluster.

FIGS. 9 A- 9 E depict hepatocyte generation from KDR expressing cells and experimental design. ( FIG. 9 A ) Immunofluorescent images showing co-expression of the endothelial cell markers KDR and CD31 in the liver of uninjured mice. ( FIG. 9 B ) Schematic structure of the Kdr-2A-Cre ERT2 -2A-eYFP construct. ( FIG. 9 C ) Representative immunofluorescent images showing specificity and sensitivity of the Kdr-2A-Cre ERT2 -2A-eYFP construct in Kdr-2A-Cre ERT2 -2A-eYFP mice. YFP expression overlaps the KDR+ expression in the liver endothelial cells. ( FIG. 9 D ) Scheme showing the experimental design for defining leakiness of Cre recombinase in Kdr-2A-Cre ERT2 -2A-eYFP, R26 LSL tdTomato mice. Mice were administered with three corn oil injections on alternate days and sacrificed 10 days later. ( FIG. 9 E ) Representative immunofluorescent microscope images showing leakiness of Cre in Kdr-2A-Cre ERT2 -2A-eYFP, R26 LSL tdTomato mice (male, n=3; female n=3). Enlarged images of selected areas demonstrate KDR+ endothelial cell identity of tdTomato+ cells.

FIGS. 10 A- 10 D depict hepatocytes in Kdr-2A-Cre ERT2 -2A-eYFP mice. ( FIGS. 10 A- 10 D ) Representative immunofluorescent microscope images of KDR+ and tdTomato+ cells and their corresponding pixel representation by ImageJ used for quantitation of percent leakiness. The bar graph depicts quantification of % tdTomato+ KDR+ cells averaged from at least four different fields in each mouse in the two groups (n=3 per group). ( FIG. 10 D ) Bar graph representing quantitation of rare tdTomato+ hepatocytes observed in Kdr-2A-Cre ERT2 -2A-eYFP, R26 LSL tdTomato mice treated with corn oil. Numerical data are presented as mean±s.d. P values were determined by two-tailed Student's t-test. ns=non-significant.

FIGS. 11 A- 11 D depict VEGFA mRNA-LNP administration inducing BEC-to-hepatocyte conversion in chronic and acute liver injury in Kdr-2A-Cre ERT2 -2A-eYFP mice. ( FIG. 11 A ) Representative image showing lineage tracing efficiency in KDR expressing cells. The labelling was uniform and complete in endothelial cells across all mice as shown. ( FIG. 11 B ) Representative immunofluorescent microscope images showing ductular response (EpCAM+ BECs) in Kdr-2A-Cre ERT2 -2A-eYFP, R26 LSL tdTomato mice treated with AAV8-Tbg-p21 vector and choline-deficient diet with 0.1% ethionine supplementation for 2 weeks, followed by VEGFA mRNA-LNP or control firefly luciferase mRNA-LNP injections, 10 μg/20 g body weight. The bar graph shows quantification of area of EpCAM+ staining averaged from at least three different fields in each mouse in the two groups (n=3 per group). ( FIG. 11 C ) Krt7 staining showing comparison of ductular response in Kdr-2A-Cre ERT2 -2A-eYFP, R26 LSL tdTomato mice treated with AAV8-Tbg-p21 vector and APAP (500 mg/kg to female mice, 300 mg/kg to male mice) followed by VEGFA mRNA-LNP or control Luciferase mRNA-LNP injections, 10 μg/20 g body weight. The bar graph represents quantification of Krt7 stained area averaged from at least three different fields in each mouse in the two groups (n=6 per group). ( FIG. 11 D ) Scheme showing the experimental design using the Kdr-2A-Cre ERT2 -2A-eYFP, mice to capture YFP (or KDR) expressing cells after acute liver toxicity. AAV8-Tbg-p21 vector was administered to induce hepatocyte senescence and injury was induced by a single intra-peritoneal injection of acetaminophen (400 mg/kg APAP to male mice and 500 mg/kg to female mice). Mice were sacrificed three days after the injury. Numerical data are presented as mean±s.d. P values were determined by one-way ANOVA followed by Tukey's method for multiple comparisons. ***p<0.001, ****p<0.0001, ns=non-significant.

FIGS. 12 A- 12 C depict histopathological examination of human liver specimens with ESLD. ( FIGS. 12 A- 12 B ) Representative brightfield images of H&E-stained cirrhotic human liver specimens showing the abnormal liver morphology and thick fibrous septa around the portal tracts of the diseased human livers. Brightfield images of trichrome stained human cirrhotic liver tissues showing collagen deposition. Oil Red O-stained human cirrhotic liver tissues showing lipid accumulation in the liver. ( FIG. 12 C ) Representative immunofluorescence images showing ductular response (KRT7+ BECs) in the tissues of human liver samples at 4× magnification. The selected regions are shown at 10× magnification to clearly visualize KRT7-positive BECs in the dense fibrous septa as depicted by DAPI staining.

FIGS. 13 A- 13 E depict evidence of BEC-to-hepatocyte conversion and KDR expression in human liver samples from non-alcoholic steatohepatitis (NASH) cirrhosis and alcoholic cirrhosis with ESLD patients. ( FIG. 13 A ) Representative immunofluorescent images of cirrhotic human liver tissues showing the GS expression in the central vein area away from the fibrous where KRT7+ BECs are located. ( FIG. 13 B ) Representative immunofluorescent images of normal human liver tissues (n=3) showing the GS expression in the central vein areas. KRT7 depicts the portal vein area. ( FIG. 13 C ) KDR and KRT7 staining showing the normal endothelial staining pattern in the diseased donor liver tissues. ( FIG. 13 D ) GS and its corresponding IgG isotype staining on serial sections demonstrating the specificity of GS antibody. ( FIG. 13 E ) Relative gene expression of albumin determined in hepatocytes isolated from normal (n=5) or cirrhotic human livers, Child Pugh B (n=5) and Child-Pugh C (n=4).

›BRIEF DESCRIPTION OF THE DRAWINGS · 4 of 10

FIG. 14 depicts a graphical summary of using VEGFA mRNA-LNP to induce liver repair. 1) In zebrafish, VEGFR signaling regulates LPC-to-hepatocyte conversion during liver regeneration in the Mtz-induced severe acute hepatocyte ablation. Chemical or genetic inhibition of VEGFR prevents cell conversion, while VEGFR overactivation via overexpression of VEGFA genetically promotes it. 2) During acute and chronic liver injuries in mice, administration of VEGFA via mRNA-LNP promotes BEC-to-hepatocyte conversion or KDR-expressing cell (mostly BECs)-to-hepatocyte conversion and reverts liver fibrosis and steatosis. Transient cytoplasmic KDR-expressing hepatocytes are detected, and are mostly descendant of KDR-expressing BECs. 3) In human ESLD cirrhotic livers, the presence of KDR-expressing BECs in septa is associated with identification of cytoplasmic KDR-expressing hepatocytes at the margin of the regenerative nodules near the septa and with transitioning hepatocytes from KRT7+GS+, KRT7−GS+, to KRT7−GS− phenotype toward the center of the nodule, strongly suggesting their BEC origin.

FIGS. 15 A- 15 B depict gating strategies of hepatocyte and non-parenchymal fractions.

FIGS. 16 A- 16 E depict liver necrosis and cell death are sexually dimorphic following administration of equivalent doses of APAP. ( FIG. 16 A ) Following the Whitten effect and fasting to normalize estrogen and glutathione levels, respectively, male and female 12-week-old C57BL/6J mice were injected with 400 mg/kg APAP intraperitoneally and followed for 96 hours using the injury scheme outlined. ( FIG. 16 B ) Serum alanine aminotransferase (ALT) levels at each sacrifice time post-APAP. First series is male, second series is female. ( FIG. 16 C ) Hematoxylin & eosin staining of representative male and female livers at each time point, shown at 40× magnification. Bar graph on the right quantifies tissue area covered by necrosis (H&E). First series is male, second series is female. ( FIG. 16 D ) TUNEL staining of representative male and female livers at each time point, quantifying both cell apoptosis and necrosis, shown at 40×. Bar graph on the right quantifies tissue area covered by cell death (TUNEL+). First series is male, second series is female. ( FIG. 16 E ) Serum bilirubin in each sex at each time point. N=3-6 mice per sex/time point, with 2-3 lobes averaged for histological quantification—one male mouse died before 96 hours. Statistics calculated via two-sided student's t test for unpaired comparisons; p-values *<0.05, **<0.001, ***<0.0001.

FIGS. 17 A- 17 E depict male and female livers exhibit distinct transcriptional responses to acetaminophen-induced liver injury. ( FIG. 17 A ) Male and female mice were given equivalent doses of 400 mg/kg APAP or PBS vehicle control post-fast and Whitten (for females), then livers were perfused in situ and dissociated at 48 hours post-APAP treatment for single-cell RNA sequencing analysis. ( FIG. 17 B ) All 4 datasets combined on a single SPRING plot (4,821 cells), with 8 different cell type clusters resolved. Hepatocytes and endothelial cells (ECs) exhibit sexually dimorphic clustering prior to APAP injury. ( FIG. 17 C ) Subclusters of combined dataset, separated by treatment, with female cells and male cells. ( FIG. 17 D ) Growth hormone receptor (GHR) expression of combined SPRING plot, and violin plots of PBS-treated and APAP-treated hepatocytes (Hep), endothelial cells (EC) showing differential expression of GHR between males and females. First series is female, second series is male. ( FIG. 17 E ) Growth hormone (GH) pathway enrichment of Biocarta gene set in SPRING and violin plots of PBS-treated and APAP-treated Heps and ECs showing differential enrichment of GH pathway between males and females. First series is female, second series is male. DEG of 0.25 used for differential expression resolution. P-values *<0.05, **<0.001, ***<0.0001.

FIGS. 18 A- 18 F depict exogenous human growth hormone treatment promotes liver recovery from APAP-induced injury and more efficiently than NAC. ( FIG. 18 A ) Injury and treatment scheme consisting of fast and Whitten effect (females), then differential severe doses of APAP (400 mg/kg for males, 600 mg/kg for females), followed by treatment of 2.5 mg/kg GH, 1000 mg/kg NAC, or equivalent volume of PBS vehicle control 8 hours after APAP injection. N=5-9 mice per sex/treatment/time point (GH and PBS) or N=3 per sex/time point (NAC), with 2-3 lobes averaged per mouse for histological quantification. ( FIG. 18 B ) Serum ALT levels of GH-treated, NAC-treated, and PBS-treated male and female mice 24 and 48 hours after severe dose of APAP. ( FIG. 18 C- 18 D ) Representative H&E stains of male mouse livers per treatment, and time point post-APAP at 40× magnification, and graph quantifying % necrotic tissue in males and females. ( FIG. 18 E- 18 F ) Representative TUNEL stains of male mouse livers per treatment, and time point post-APAP at 40× magnification and graph quantifying % TUNEL+ tissue in males and females. Statistics calculated via two-sided student's t test for unpaired comparisons; p-values *<0.05, **<0.001, ***<0.0001.

FIGS. 19 A- 19 D depict constitutive activation of the downstream GH/GHR pathway mediator STAT5b rescues males from APAP hepatotoxicity. ( FIG. 19 A ) Pre-treatment scheme with AAV8-TBG-STAT5bCA given 7 days prior to 400 mg/kg severe APAP injury, in C57BL/6J male mice, and sacrificed at 48 hours post-APAP administration. ( FIG. 19 B ) Plasma murine IGF1 concentration measured by ELISA 6 days post-AAV8 injection in Null- and STAT5bCA-injected mice. ( FIG. 19 C ) Survival rate for STAT5bCA- (square) and Null-injected mice (x). ( FIG. 19 D ) Representative H&E stain and TUNEL stain of STAT5bCA-injected mouse livers, with average of % necrotic tissue and % TUNEL+ cell area and standard error for treatment group. N=6 mice/treatment. Statistics calculated via two-sided student's t test for unpaired comparisons; p-values *<0.05, **<0.001, ***<0.0001.

›BRIEF DESCRIPTION OF THE DRAWINGS · 5 of 10

FIGS. 20 A- 20 F depict GH mRNA-LNP targets liver to promote recovery throughout the length of the APAP-induced acute injury. ( FIG. 20 A ) C57BL/6J mice were given sub-lethal doses of APAP (400 mg/kg for males, 600 mg/kg for females), and 8 hours later were injected retro-orbitally with g of human GH mRNA-LNP or negative control Luc mRNA-LNP, then analyzed 24 and 48 hours post-APAP. ( FIG. 20 B ) Upper panels: human GH protein levels in liver tissue homogenate (left) and serum (right) were measured 5 hours post-injection by ELISA. Lower panel: human GH protein levels in serum of mice were measured 5, 24 and 48 hours post GH mRNA-LNP injection by ELISA. N=3 mice per treatment/time point. ( FIG. 20 C ) Survival rate of GH mRNA-LNP-treated mice (squares; males in blue, females in pink) and Luc mRNA-LNP-treated mice (black x's) 24 and 48 hours post APAP overdose. N=5 mice per sex/treatment/time point. Statistical analysis carried out using the Log-rank Mantel-Cox test. ( FIG. 20 D ) Serum ALT of GH mRNA-LNP-treated females and males and Luc mRNA-LNP-treated mice 24 and 48 hours post APAP treatment. Note all male mice treated with Luc mRNA-LNP died by 48 hours post treatment (skull symbol). The first series in each timepoint is Luc mRNA-LNP. The second series in each timepoint is GH mRNA-LNP. ( FIG. 20 E ) % necrotic liver tissue quantified from H&E stains in GH mRNA-LNP-treated and Luc mRNA-LNP-treated mice. The first series in each timepoint is Luc mRNA-LNP. The second series in each timepoint is GH mRNA-LNP. ( FIG. 20 F ) % TUNEL+ area from TUNEL stain in GH mRNA-LNP-treated and control Luc mRNA-LNP-treated mice. The first series in each timepoint is Luc mRNA-LNP. The second series in each timepoint is GH mRNA-LNP. Statistics for bar graphs calculated via two-sided student's t test for unpaired comparisons; p-values *<0.05, **<0.001, ***<0.0001.

FIG. 21 A- 21 C depict Male and female hepatocytes and endothelial cells are transcriptionally distinct. ( FIG. 21 A ) Single liver hepatocytes (Hep) and endothelial cells (EC) from males and females treated with PBS or APAP harvested as described in FIG. 17 were analyzed for pathway enrichment using ENRICHR. From the SPRING plots, the list of the most enriched genes in pre-identified hepatocytes and endothelial cells in each group were defined, analyzed with ENRICHR for pathway analysis, and ranked by z-score of enrichment as defined in the Bioplanet 2019 pathways database. B/C: Violin plots from Heps and ECs of genes included in the BioCarta GH pathway activation ( FIG. 21 B ) as well as of additional downstream genes key for GH pathway activation ( FIG. 21 C ). T-tests were used to assess differences in gene set scores between groups for each cell type, p-values *<0.05, **<0.001, ***<0.0001.

FIGS. 22 A- 22 E depict Median dosage of 2.5 mg/kg GH is most beneficial for treatment post APAP injection. ( FIG. 22 A ) Serum ALT, % necrotic liver tissue and % TUNEL+ liver tissue quantified from H&E stain and TUNEL stain, respectively, from males and females 12 hours post-APAP treatment with doses of 400 mg/kg and 600 mg/kg, respectively. First series is male, second series is female. ( FIG. 22 B ) Injury and treatment scheme for male and female mice injured with sex-specific doses of APAP and treated 8 hours later with 4 different doses of subcutaneous GH protein injection or PBS vehicle control, and sacrificed at either 24 hours post-treatment (females) or 48 hours post-treatment (males). ( FIG. 22 C ) Serum ALT levels of GH-treated and PBS-treated male and female mice 24 and 48 hours after administration of APAP. First series is male, second series is female. ( FIG. 22 D ) % Necrotic liver tissue quantified via H&E stain for males and females following PBS or GH protein treatment at 4 different doses. First series is male, second series is female. ( FIG. 22 E ) Representative H&E images for each sex at each GH dosage shown at 40× magnification. Statistics calculated via two-sided student's t test for unpaired comparisons; p-values *<0.05, **<0.001, ***<0.0001.

FIGS. 23 A- 23 C depict GH treatment promotes recovery to a lesser extent in females. ( FIGS. 23 A- 23 B ) Pictures of livers from female mice described in FIG. 18 treated with APAP, NAC or PBS. Representative H&E stains ( FIG. 23 A ) or TUNEL stains ( FIG. 23 B ) from mice used in FIG. 17 of livers per treatment, and time point post-APAP at 40× magnification. ( FIG. 23 C ) Survival rate calculated from GH-treated (squares) vs NAC-treated (black x's) mice after APAP overdose. N=6 mice per sex/treatment. Statistical analysis carried out using the Log-rank Mantel-Cox test.

FIG. 24 depicts experimental design of the hiPSC-based bi-cell therapy with engineered HLCs and ECs to treat liver disease models. Aim 1 design will enhance EC supportive functions by engineering ECs to express VEGFR2 and the downstream factors HGF, WNT2, and WNT9b, while the ligand VEGFA will be delivered via mRNA-LNPs. Aim 2 design will promote maturation and engraftment of HLCs that will be engineered to express mitogen receptors and maturation transcription factors (TFs), while mitogens will be delivered via mRNA-LNPs. Experiments in both aims will establish novel engineered hiPSC lines in which genetic circuits will sense ECs via CD31 or CD144 expression, and HLCs via FOXA2 or AFP expression as the cells specify in culture, and subsequently will induce expression of the cell type-specific genes.

FIGS. 25 A- 25 D depicts efficient generation of HLCs in 2D hiPSC differentiation cultures. ( FIG. 25 A ) Timeline of hiPSC 2D differentiation. ( FIG. 25 B ) Day 5 cultures generate −100% of endoderm cells expressing CXCR4 and CKIT that in turn give rise to a homogenous population of HLCs at day 15 expressing all AFP and EPCAM while 25% express AAT. ( FIG. 25 C ) qPCR quantification and ( FIG. 25 D ) immunostaining for hepatic markers in day 15 HLCs.

FIGS. 26 A- 26 F depicts integration of day 17 hESC-derived HLCs in CCI4-NSG mice ( FIGS. 26 A- 26 D ) and FRG mice ( FIGS. 26 E- 26 F ). ( FIG. 26 A ) Bioluminescence pictures and ( FIG. 26 B ) bioluminescence quantification representing transplanted cells in livers at T1, T3 and T7. ( FIG. 26 C ) Immunostaining of clusters of human albumin+ cells at T7 (arrows). ( FIG. 26 D ) ALT serum levels in mice at T-7 (7 days prior to transplantation), and after transplantation at T1, T3 and T7. N=3-5 mice per group. ( FIG. 26 E ) Protocol of cell transplantation in FRG mice. ( FIG. 26 B ) Presence of human albumin+ cell clusters 40 days after transplantation.

›BRIEF DESCRIPTION OF THE DRAWINGS · 6 of 10

FIGS. 27 A- 27 C depicts mRNA-LNPs are efficiently targeted to the liver. ( FIG. 27 A ) Bioluminescence pictures of mice injected with 5 ug of luciferase-mRNA-LNP. ( FIG. 27 B ) Luciferase activity (photon/sec) seen in ( FIG. 27 A ) in liver areas. ( FIG. 27 C ) GFP immunofluorescence in liver 5 hours after injection of GFP-mRNA-LNPs. Note that all HNF4α+ hepatocytes express GFP.

FIGS. 28 A- 28 F depicts mRNA-LNP encoding HGF, EGF, and IL6 efficiently promote hiPSC-derived HLC survival in acute (APAP: FIGS. 28 A, 28 B, 28 E, and 28 F ) and chronic (PiZ; FIGS. 28 C and 28 D ) liver injury models. Mice were either injected with rnRNA-LNP encoding HGF, IL6, and EGF (mRNA), or PolyC control, and then 10 6 BU3 hiPSC-derived HLCs were transplanted through the spleen of all mice 5 hours later. The following day, mice were analyzed for bioluminescence activity. ( FIGS. 28 A, 28 C ) Pictures taken 10 min after luciferin injection and ( FIGS. 28 B, 28 D ) luciferase activity in the respective liver areas quantified (circled areas). Note the greater luciferase activity in all mice treated with mRNAs in both liver injury mouse models. ( FIGS. 28 E, 28 F ) Presence of single HLCs expressing human nuclei antigen ( FIG. 28 E ) or cluster of HLCs ( FIG. 28 F ) detected with DsRed 1 day after cell transplantation in the APAP injury model.

FIGS. 29 A- 29 C depict validation of activation of genetic circuits inducing gene express (actuator: tdTomato) in cells that sense specific gene expression (sensor: RSP21/EYFP) using the CRISPR/dCas9 system. ( FIG. 29 A ) Design of the sensor/actuator circuit. ( FIG. 29 B ) Activity of the circuit does not affect the endogenous expression of the sensor gene RPS21/EYFP. ( FIG. 29 C ) Expression of the actuator tdTomato is proportional to the number of gRNAs integrated chromosomally downstream of endogenous RPS21 gene.

FIGS. 30 A- 30 B depict design of the sensor/actuator genetic circuits in hiPSC lines to express either VEGFR2 ( FIG. 30 A , line 1) or HGF/WNT2/WNT9b ( FIG. 30 B , line 2) in CD31 or CD144 (sensor) expressing ECs. Line 3 will express VEGFR2 as well as the 3 endothelial factors in either CD31 or CD144 expressing ECs.

FIG. 31 depicts APAP-NSG liver injury model. 200-400 mg/Kg doses of APAP were tested for centrilobular necrosis 24 h after injection. Gross morphology of the liver as well as H&E staining illustrate the increase liver damage with increasing doses. 400 mg/kg dose triggered severe damage and ultimately death, while 200 mg/Kg-mediated damage was mild.

FIGS. 32 A- 32 B depict design of the sensor/actuator genetic circuit in hiPSC line to induce robust expression of either cMET, EGFR and IL6R in hiPSC line 1 (A) and PROX1, CEBPA and ATF5 in hiPSC line 2 (B) in AFP or FOXA2 (sensor) expressing HLCs. A line 3 is also established that will generate HLCs that will express the 3 receptors using FOXA2 expression as sensor as well as the 3 maturation TFs using AFP expression as sensor.

FIGS. 33 A- 33 B depict design of bi-cell transplantation and mRNA-LNP conditions. ( FIG. 33 A ) Bi-cell transplantation design and analyses. ( FIG. 33 B ) Ligand-mRNA-LNP conditions to be tested.

FIGS. 34 A- 34 B depict proposal overview. ( FIG. 34 A ) Current primary human hepatocytes (PHH) and HLC therapies have limitations. ( FIG. 34 B ) It is contemplated that engineering transplanted HLCs to express important dox-inducible mitogen receptors and maturation transcription factors and advantaging transplanted cells with mRNA-LNP will improve HLC repopulation in the NSG-PiZ mouse model recapitulating alpha 1 antitrypsin disease (AATD) associated liver disease.

FIGS. 35 A- 35 B depict NSG-PIZ mice recapitulate AATD liver phenotype. ( FIG. 35 A ) Z-AAT polymer accumulation in hepatocytes visualized as PAS-positive diastase-resistant. ( FIG. 35 B ) Hepatic fibrosis as shown with picrosirius red stain.

FIGS. 36 A- 36 D depict Wildtype PHHs repopulate the NSG-PiZ mouse liver. ( FIG. 36 A ) Serum human albumin levels quantified by ELISA biweekly post-engraftment for 10 weeks following transplantation. ( FIG. 36 B ) Human albumin immunostaining on liver sections 10 weeks post-partial hepatectomy (PHx). ( FIG. 36 C ) Serum human albumin after various challenges. ( FIG. 36 D ) Serum human AAT levels quantified by ELISA at 10 weeks post-engraftment. Dotted line is therapeutic threshold, 572 mg/mL.

FIGS. 37 A- 37 D depict HLCs have immature gene signature. ( FIG. 37 A ) Schematic of 15 day directed differentiation from iPSC to HLC. ( FIG. 37 B ) Flow cytometry analyses of day 5 definitive endoderm culture. ( FIG. 37 C ) Immunofluorescence of cells in culture throughout differentiation. ( FIG. 37 D ) RT-qPCR analysis showing relative gene expression during differentiation. Fold change is relative to PHH control. Data points are mean±SEM, n=1.

FIGS. 38 A- 38 E depict mRNA-LNP express proteins in the liver. ( FIG. 38 A ) eGFP detected 5 h after mRNA-LNP injection in NSG-PiZ mice. ( FIG. 38 B ) NSG-PiZ mice were treated with or without intravenous AAV8-P21. 7 d later, they were injected with HGF+EGF mRNA-LNP or PolyC control RNA-LNP. 48 h later, mice were injected with EdU to label proliferating cells. Livers were harvested 2 h later. ( FIG. 38 C ) EdU stain showing proliferating cells in green. ( FIG. 38 D ) Quantification of EdU+ hepatocytes per 10× image field. n=2 or 3 mice. Data represented as mean±SEM. ( FIG. 38 E ) Immunofluorescence showing P21 expression in AAV8-P21 treated NSG-PiZ mice 1 week post injection and in P21 mRNA-LNP treated NSG mice 5 hrs post injection.

FIG. 39 depicts HGF+EGF mRNA-LNP increases cluster size of PHHs in NSG-PiZ male mice. Mice were transplanted with 10 6 PHHs. PolyC or HGF+EGF mRNA-LNP injections were given once a week post transplantation. Livers were harvested after 9 weeks. hALB shows PHHs, n=1.

FIGS. 40 A- 40 C depict HGF+EGF mRNA-LNP transiently improves survival of HLCs in NSG-PiZ mice. ( FIG. 40 A ) Schematic of in vivo detection of transplanted HLCs. ( FIG. 40 B ) NSG-PiZ mice were treated with either PolyC or HGF+EGF mRNA-LNP 5 h prior to transplantation with 10 6 HLCs. Representative images of bioluminescence detected with in vivo imaging system (IVIS) 1, 3, and 5 days post transplantation. ( FIG. 40 C ) Quantification of IVIS data represented as mean±SEM, n=5-6 per group.

›BRIEF DESCRIPTION OF THE DRAWINGS · 7 of 10

FIG. 41 depicts PiggyBac based non-viral gene delivery of doxycycline-inducible genetic constructs.

FIG. 42 depicts mechanisms of action of the VEGFA-VEGFR2 axis on BEC-to-hepatocyte conversion and restoration of liver function in mouse and zebrafish models. BEC: biliary epithelial cells, EC: endothelial cells, LPC: liver progenitor cells.

FIG. 43 depicts schematic of the process of BEC-driven liver regeneration in the zebrafish NTR-mediated hepatocyte ablation model. Tg(fabp10a:CFP-NTR) larvae were treated with 10 mM Mtz from 3.5 days post-fertilization (dpf) for 36 hours, followed by Mtz washout. A0h stands for ablation 0 hour; R0h, regeneration 0 hour; R48h, regeneration 48 hours.

FIG. 44 A- 44 C depict VEGFR2 inhibition impairs BEC-driven liver regeneration in zebrafish. ( FIG. 44 A ) Epifluorescence images showing fabp10a:DsRed expression in the regenerating larvae at R24h and R48h. Arrows point to the liver. Quantification of the liver size is shown. ( FIG. 44 B ) Confocal images showing Prox1 and Hnf4a expression in regenerating livers at R6h. ( FIG. 44 C ) Confocal images showing the expression of fabp10a:CFP-NTR, Tp1:H2B-mCherry, and Bhmt in regenerating livers at R24h. Scale bars, 50 μm.

FIGS. 45 A- 45 G depict the oncogene-induced liver injury model of LPC-mediated liver regeneration, Tg(fabp10a:ca-β-catenin). ( FIG. 45 A ) Whole-mount in situ hybridization images showing the expression of senescence markers, tp53 and cdkn1a, at 5 dpf ( FIG. 45 B ) SA-β-gal staining also reveals the increased senescence in Tg(fabp10a:ca-13-catenin) livers at 10 dpf Arrows point to livers. ( FIG. 45 C ) Confocal images showing the labeling of TUNEL and nucleus (Hoechst33342) with fabp10a:GFP expression at 10 dpf TUNEL + cells (arrow) were detected in Tg(fabp10a:ca-13-catenin) livers but not in control livers. ( FIG. 45 D- 45 F ) Confocal images showing the expression of mpeg1:Dendra2 (macrophages; FIG. 45 D ), hand2:EGFP (stellate cells; FIG. 45 E ), and acta2:mCherry (fibrosis; FIG. 45 F ). Anxa4 is a marker of BECs and LPCs. ( FIG. 45 G ) Confocal images showing the expression of fabp10a:CFP (hepatocytes and LPCs), Tp1:H2B-mCherry (BECs and LPCs), and Bhmt (hepatocytes) in Tg(fabp10a:ca-13-catenin) livers. Arrows point to LPCs. Scale bars: 50 μm.

FIGS. 46 A- 46 E depict mRNA-LNPs are efficiently transfected in virtually all hepatocytes. ( FIG. 46 A ) Bioluminescence pictures of mice injected IV with 10 μg of luciferase mRNA-LNP. ( FIG. 46 B ) Graph representing the luciferase activity (photon/sec) seen in ( FIG. 46 A ). ( FIG. 46 C ) flow cytometry analyses of dissociated liver cells and ( FIG. 46 D ) GFP expression on liver sections 5 hours after injection of GFP mRNA-LNP. ( FIG. 46 E ) Efficient VEGFA secretion in serum with time assessed by ELISA specific for human VEGFA after injection of 10 μg of human VEGFA mRNA-LNP (mean±SD from 3 mice/time point).

FIG. 47 depicts DR develops in both chronic CDE diet and acute APAP liver injury in the presence of AAV8-Tbg-p21. Expansion of KRT7+ BECs from the portal triads is greater in the CDE model compared to the APAP model. *Central vein areas.

FIG. 48 A- 48 C depict VEGFA induces BEC-to-hepatocyte conversion in CDE-fed/p21-injected mice. ( FIG. 48 A ) Experimental design using the KRT19-Cre ERT , R26 LSL tdTomato mice. ( FIGS. 48 B, 48 C ) Tomato and KRT7 immunostaining on either Poly(C)RNA-LNP ( FIG. 48 B , n=4) or VEGF mRNA-LNP ( FIG. 48 C , n=4) treated mice 5 days following the last injection of mRNA-LNPs. Note the numerous areas of Tomato+ hepatocytes (arrowheads) adjacent to Tomato+ KRT7+ BECs (arrows) in VEGFA-treated mice. * represent Tomato+ BEC areas.

FIG. 49 A- 49 D depict VEGFA reverts fibrosis and steatosis in CDE-fed/p21-injected mice. Trichrome and lipidspot assays were performed ( FIGS. 49 A, 49 B ) and quantified ( FIGS. 49 C, 49 D ) on mice treated with either Poly(C) RNA-LNPs ( FIG. 49 A , n=4) or VEGFA mRNA-LNPs ( FIG. 49 B , n=4).

FIGS. 50 A- 50 D depict VEGFA induces BEC-to-hepatocyte conversion in APAP/p21 treated mice. ( FIG. 50 A ) Experimental design using the KRT19-Cre ERT , R26 LSL tdTomato mice. ( FIG. 50 B ) Tomato and HNF4α immunostaining on Poly(C) RNA-LNP or VEGFA mRNA-LNP treated mice 5 and 14 days following the last injection of mRNA-LNPs. ( FIG. 50 C ) Flow cytometry quantification of CD26+ Tomato+ hepatocytes, and ( FIG. 50 D ) corrected based on the Krt19 BEC lineage tracing efficiency as measured by the percentage of Tomato+ EpCAM+ BEC for each mouse (n=4 mice/group).

FIG. 51 depicts experimental designs of acute and chronic liver mouse injuries.

FIG. 52 depicts analyses of VEGFA effects on BEC characteristics, BEC expansion (stage 1) and emergence/proliferation of LPCs and KDR+ BEC (phase 2), BEC-to-hepatocyte conversion (phase 3) and subsequent de novo hepatocyte proliferation (phase 4).

FIG. 53 depicts scheme of VEGFA overexpression experiments. Tg(fabp10a:ca-β-catenin); Tg(hsp70l: loxP-mCherry-loxP-VEGFA); Tg(Tp1: CreERT2); Tg(ubb:loxP-CFP-STOP-loxP-H2B-mCherry) quadruple transgenic larvae will be treated with 10 μM 4-OHT from 3 to 4 dpf and heat-shocked (HS) twice at 13 and 14 dpf. The larvae will be harvested at 15 or 30 dpf for liver analysis.

FIG. 54 depicts proposed genetic zebrafish models associated with the slow or fast BEC-driven regeneration models.

FIGS. 55 A- 55 C depict expression of VEGFR2 in ECs ( FIG. 55 A ) and BECs ( FIGS. 55 B, 55 C ) from the CDE diet ( FIGS. 55 A, 55 B ) and the APAP ( FIG. 55 C ) induced liver injuries in the presence of AAV8-Tbg-p21. Detection of KRT19+ or KRT7+ BECs positive for VEGFR2 (arrowhead) in CDE ( FIG. 55 B ) and APAP ( FIG. 55 C ) models.

FIGS. 56 A- 56 D depict validation of the Kdr-2A-Cre ERT2 -2A-eYFP mouse.

FIGS. 57 A- 57 B depict the PI3K-AKT-mTORC1 axis controls BEC-driven liver regeneration. ( FIG. 57 A ) Confocal images showing the expression of fabp10a:CFP-NTR, Tp1:H2B-mCherry, and Bhmt in regenerating livers at R24h. 10 μM LY294002 was added from A18h to R24h8. ( FIG. 57 B ) Confocal images showing pS6 expression in 5-dpf control and R6h regenerating livers. Arrows point to pS6-positive BECs; arrowheads to pS6-positive hepatocytes. Scale bars, 50 μm.

›BRIEF DESCRIPTION OF THE DRAWINGS · 8 of 10

FIG. 58 depicts generation of genetic tools for AKT activation. Confocal images showing the expression of TRE:CFP, TRE:myrAkt1-P2A-Venus, and pS6 in the liver at 5 dpf. pS6 expression was observed in TRE:Venus + hepatocytes (arrows). The larva was treated with 10 μg/ml Dox from 4 to 5 dpf. Scale bar, 50 μm.

FIG. 59 depicts mechanisms of action of GH pathway on susceptibility and recovery from APAP-induced liver injury. Aim 1 will test whether sex-specific GH level disparity is a key determinant of the susceptibility of APAP-induced injury and subsequent repair. Aim 2 will establish an optimum therapy to treat APAP-induced liver damage by injection of GH, IGF1, and GH/IGF1 vs the standard-of-care NAC therapy. Aim 3 will identify the molecular and cellular actions of GH/IGF1 therapy to promote liver repair after APAP overdose.

FIGS. 60 A- 60 G depict liver necrosis, hepatocyte apoptosis and tissue recovery are sexually dimorphic following equivalent doses of APAP. ( FIG. 60 A ) Male and female mice were injected with 400 mg/kg APAP and monitored for 96 hours using the injury scheme outlined. ( FIG. 60 B ) Serum ALT levels with time in both sexes (each dot represents one mouse). ( FIG. 60 C ) H&E staining at 24, 48 and 96 hours post-APAP shows less necrosis in females overtime. (100× magnification). ( FIG. 60 D ) Graph quantifying surface area covered by necrotic (eosin+/hematoxylin−) tissue in each sex over time (n=4 mice/sex/time point, 2 lobes per mouse). ( FIG. 60 E ) TUNEL staining representing cell apoptosis is consistent with H&E pattern (40×). ( FIG. 60 F ) Graph quantifying surface area covered by TUNEL+ staining (n=4 mice/sex/time point). ( FIG. 60 G ) Quantification of serum bilirubin (n=4 mice/sex/time point). Two tailed t-test, p-values *<0.05, **<0.001, ***<0.0001, n.s.=not significant.

FIG. 61 depicts SPRING plot of scRNA Seq analyses of liver cells from APAP- and PBS-treated males and females. 10 putative cell types are resolved.

FIGS. 62 A- 62 G depict APAP triggers greater activity of GHR in female hepatocytes and ECs compared to their male counterparts. ( FIG. 62 A- 62 F ) SPRING plots of combined cells from APAP- and PBS-treated males and females illustrate expression for GHR, IGF1, IGF1R, IGFBP4, IGFBP7 and prolactin receptor (PRLR). The intensity represents levels of gene expression. Bars on violin plots represent average expression level in hepatocytes (Hep), ECs and KC/monocytes (M/KC). First series in each graph is female, second series is male. ( FIG. 62 G ) Biocarta GH pathway expression is significantly higher in female hepatocytes and ECs than in male cell counterparts. Pathways with normalized enrichment scores of p>0.2 shown. Welch's two sample t-test was used to calculate statistical significance (***p<0.0001, **p<0.001, *p<0.05, n.s.: not significant).

FIGS. 63 A- 63 F depict a single injection of GH sharply reduces injury and accelerates liver regeneration following similar level of APAP-induced liver damage in both sexes. ( FIG. 63 A ) Timeline of the experimental design. ( FIG. 63 B ) Serum ALT levels over time in both sexes (each dot represents one mouse). ( FIG. 63 C ) H&E stain of male and female livers given either PBS or GH 8 hours after sub-lethal sex-dependent APAP injection dosing, then euthanized 24 h or 48 h post-APAP (40× magnification). ( FIG. 63 D ) Quantification of staining shown in C by measuring stained area on FIJI (n=3 mice/sex/treatment/time point, 2 lobes per mouse). First series is untreated, second series is treated. (FIG. 63 E) TUNEL staining on subsequent tissue sections from the same mice (40×). ( FIG. 63 F ) Quantification of staining shown in E by measuring stained area on FIJI. (n=3 mice/sex/treatment/time point, 2 lobes per mouse). First series is untreated, second series is treated. Two tailed t-test, p-values *<0.05, **<0.001, ***<0.0001, n.s.=not-significant.

FIGS. 64 A- 64 D depict mRNA-LNPs are efficiently transfected in virtually all hepatocytes and a subpopulation of ECs and macrophage/KCs 1 . ( FIG. 64 A ) Bioluminescence pictures of mice injected IV with 10 μg of luciferase mRNA-LNP. ( FIG. 64 B ) Graph representing the luciferase activity (photon/sec) seen in ( FIG. 64 A ). ( FIG. 64 C ) eGFP expression on liver sections 5 hours after injection of eGFP mRNA-LNP, ( FIG. 64 D ) flow cytometry analyses of dissociated liver cells (Poly(C) RNA-LNP used as control, mean±SD from 2 mice/group measured in duplicate). 1

FIG. 65 depicts timeline of the experimental design to investigate the phenotypic sexual disparity of GH levels in APAP susceptibility to liver injury and repair.

FIG. 66 depicts strategy of scRNA Seq analyses to investigate the role of sex-differential GH profiles in APAP susceptibility to liver injury and repair. For each cell type identified, the 4 transcriptomes (female, male, and cGH or PBS minipump treated male (Male-GH or Male-PBS) will be analyzed for cell cycle, GSEA, specific Hallmark/Biocarta gene set related to GH pathway. They will be compared to obtain the set of transcripts related to female GH level-mediated protection/repair (green), as well as the set of transcripts related to female-specific protection/repair, excluding genes related to GH-mediated protection/repair (purple) (see Venn diagram). Cell-cell crosstalk will be identified using the CellPhoneDB algorithm.

FIGS. 67 A- 67 E depicts identification from the scRNA Seq data early and late estrogen pathway 3 as a candidate female-specific pathway whose activation is associated with resistance to APAP liver injury. ( FIG. 67 A- 67 C ) SPRING plots and violin plots for expression of ERa and of early and late estrogen pathway. ( FIG. 67 D ) Timeline of E2 treatment in male mice. ( FIG. 67 E ) Serum ALT levels in control oil-treated and E2-treated males (each dot represents one mouse). Two tailed t-test, p-values *<0.05, n.s: not significant. Hep: hepatocyte, EC: endothelial cells.

FIG. 68 depicts experimental design for comparison of efficacy of GH/IGF1 treatment using recombinant protein or mRNA-LNP vs clinical standard-of-care NAC post-APAP.

›BRIEF DESCRIPTION OF THE DRAWINGS · 9 of 10

FIGS. 69 A- 69 B depicts experimental design prior to collection of liver cell suspensions for scRNA sequencing analyses and liver function assays ( FIG. 69 A ), as well as for cell-specific knock-out models for which examples are shown ( FIG. 69 B ).

FIG. 70 depicts chronic liver disease is a critical health burden in the USA. NAFLD is the first stem of chronic liver disease and can eventually lead into irreversible cirrhosis.

FIG. 71 depicts strategies to harness liver regeneration. Strategies include hepatocyte-driven liver repair in which hepatocytes proliferate to compensate for loss of liver tissue; cholangiocyte-driven liver repair when hepatocyte proliferation is exhausted is case of chronic or acute injury; and cell therapy using primary human hepatocytes and patient specific induced pluripotent stem cell derived hepatocytes. To do so HGF, EGF, and VEGFA can be utilized to promote hepatocyte survival and proliferation and to activate liver progenitor cells to produce de novo functional hepatocytes.

FIGS. 72 A- 72 C depict intravenous injection of nucleoside modified mRNA encoding luciferase complexed to lipid nanoparticles target the liver specifically.

FIGS. 73 A- 73 B depict cells transfected by mRNA-LNP encoding enhanced GFP. After intravenous injection of eGFP mRNA-LNP, virtually all HNF4α+ hepatocytes were transfected, while cholangiocytes, the CK19 positive cells were not. In addition to hepatocytes, about 70% of CD31+ endothelial cells and 40% s of the F4/80 Kupffer cells, the macrophages of the liver, were also transfected. This was quantified by flow cytometry after dissociation of the liver.

FIGS. 74 A- 74 B depicts HGF mRNA-LNPs induce hepatocyte proliferation in homeostasis condition. 5 hours after HGF mRNA-injection, HGF expression was very intense in all hepatocytes. Hepatocyte proliferation was seen 24 h00 and 48H00 after injection as assessed with staining of incorporated Edu. Quantification shows that more than 60% of the Edu+ cells are hepatocytes in the HGF treated group and that the number of proliferative hepatocytes was 121 fold greater in this group compared to numbers found in the control PolyC treated group.

FIGS. 75 A- 75 C depict HGF and EGF mRNA-LNPs revert liver necrosis and reduce steatosis in a chronic liver injury model. Mice were fed the choline deficient ethionine supplemented CDE diet for 3 weeks to induce steatohepatitis and hepatocyte death. Mice were then treated twice with HGF EGF mRNA LNP during the recovery period. The liver mitogen mRNA-LNP rapidly restores the serum ALT levels indicative of liver damage to normal levels 2 days after the first injection. The treatment also induced a sharp decrease of steatosis compared to that with control Poly(C)-RNA treatment, in which macrosteatosis was maintained, even 8 days after the diet was over. The levels of cholesterol found in the serum of mice were inversely proportional to the degree of steatosis seen in livers.

FIGS. 76 A- 76 C depicts HGF and EGF mRNA-LNPs accelerate liver repair following acetaminophen-induced acute liver injury. A single injection of HGF and EGF mRNA-LNP was administered 24 hours after acetaminophen overdose, and mice were analyzed 8 hours and 24 hours after the mRNA-LNP injection. Beneficial effects of mRNA-LNP were consistently and significantly observed 24 hours after HGF/EGF mRNA-LNP injection with accelerated disappearance of necrotic areas that were still seen in the Poly(C) RNA-LNP-treated control group assessed by H&E staining, which was accompanied by significantly lower serum ALT levels, and absence of TUNEL cells that were still present in Poly(C) RNA-LNP control group.

FIG. 77 depicts use of HGF/EGF mRNA-LNPs to harness hepatocyte-driven liver repair by transiently promoting hepatocyte proliferation and reverting steatosis in a chronic liver injury model.

FIGS. 78 A- 78 C depict VEGFA mRNA-LNPs promote cholangiocyte-to-hepatocyte conversion in vivo and reverse the chronic liver disease. Lineage tracing of cholangiocytes in a CDE-induced chronic liver disease mouse model treated with 2 doses of VEGFA mRNA-LNP after the CDE diet revealed VEGFA significantly induces generation of tomato+ hepatocytes throughout the liver.

FIGS. 79 A- 79 B depict VEGFA mRNA-LNPs promotes cholangiocyte to hepatocyte conversion and reverses steatosis and fibrosis in a chronic liver injury model. Microvesicle of fat are seen in hepatocytes in control PolyC treated mice, while these microvesicles are absent in the VEGFA mRNA-LNPs treated mice. Similarly, VEGFA mRNA-LNPs reverted fibrosis as assessed with trichrome staining. Fibrosis is still present in polyC treated mice, while it is absent in VEGFA mRNA-LNPs treated mice.

FIG. 80 depicts mechanisms of hepatocyte regeneration and the proposed experiments to promote cholangiocyte-driven liver regeneration by activating liver progenitor cells (LPCs) to differentiate into hepatocytes using VEGFA, a ligand for VEGFR2/KDR.

FIGS. 81 A- 81 B depict the identification of liver progenitors expressing VEGFR2/KDR.

FIG. 82 depicts the experimental mouse models and design to test if VEGFA, a ligand for VEGFR2/KDR promotes cholangiocyte to hepatocyte conversion.

FIG. 83 depicts liver injury induces KDR expression on cholangiocytes.

FIG. 84 depicts mapping hepatocytic fate of cholangiocytes and KDR expressing cells in VEGFA-treated mice.

FIGS. 85 A- 85 E demonstrate VEGFA mRNA-LNPs promotes cholangiocyte to hepatocyte conversion and reverses steatosis and fibrosis in a chronic liver injury model.

FIGS. 86 A- 86 D demonstrate that VEGFA mRNA-LNPs promotes KDR+ cell conversion to hepatocytes in acute and chronic liver injuries.

FIGS. 87 A- 87 F summarize that KDR is expressed on a subset of cholangiocytes after acute and chronic liver injuries and that VEGFA mRNA-LNPs promotes cholangiocyte to hepatocyte conversion in acute and chronic liver injuries.

FIGS. 88 A- 88 B depict NSG-PiZ mice recapitulate alpha-1 antitrypsin deficiency liver disease.

FIG. 89 depicts alpha-1 antitrypsin deficiency is an ideal candidate for liver cell therapy.

›BRIEF DESCRIPTION OF THE DRAWINGS · 10 of 10

FIG. 90 depicts experimental proposal to use mRNA-LNP encoding HGF and EGF to stimulate proliferation of transplanted cells in NSG-PiZ mouse model.

FIGS. 91 A- 91 B depict a single IV injection of mRNA-LNP induces robust and restricted protein expression in the liver.

FIGS. 92 A- 9 B show that hepatocytes are the main liver cell type transfected by mRNA-LNP.

FIG. 93 shows that mRNA-LNP induces protein expression in diseased hepatocytes in NSG-PiZ mice.

FIG. 94 shows HGF+EGF mRNA-LNP induces hepatocyte proliferation.

FIG. 95 shows that weekly injections of HGF+EGF mRNA-LNP increase PHH cluster size in male NSG-PiZ mice.

FIGS. 96 A- 96 C show that HGF+EGF mRNA-LNP transiently improves HLC survival after transplantation.

FIGS. 97 A- 97 E show that in equivalent doses of APAP (400 mg/kg), the level and persistence of tissue necrosis and apoptosis are highly sexually dimorphic (n=4 mice/sex/time point). P<0.05=*, <0.005=**, <0.0005=***. In FIG. 97 A- 97 C the first series is male and the second series is female.

FIGS. 98 A- 98 C show single-cell RNA sequencing of whole livers of male and female mice before and after injury reveals sexual dimorphism of growth hormone (GH) pathway activity in hepatocyte and endothelial cell transcriptomes. Data are shown on SPRING plots of combined populations, and violin plots of hepatocyte and endothelial clusters after APAP injury dissecting differences between sexes. Lines on violin plots represent mean expression level for cluster.

FIGS. 99 A- 99 E show that a single dose of GH significantly accelerates liver regeneration after APAP-induced liver injury in both males and females, as compared to PBS-treated and NAC standard of care-treated controls. (n=5 mice/sex/treatment). Females and males were treated with sex-specific doses of APAP that generated similar liver damage. First series is untreated, second series is treated.

FIGS. 100 A- 100 B show a single injection of nucleoside-modified lipid nanoparticle-encapsulated mRNA encoding GH induces robust expression of GH in the liver and serum of uninjured mice 5 hours post retroorbital injection, as compared to Luciferase mRNA-LNP (Luc) negative control. Serum concentration of GH is 30× less than acute recombinant GH injection, within safe limit for clinical application. Future treatment model will test the efficacy of promoting recovery from APAP injury compared to recombinant protein injection. (n=3 mice/treatment).

FIG. 101 depicts alpha-1 antitrypsin deficiency (AATD) increases risk for liver and lung disease.

FIG. 102 depicts NSG-PiZ mice recapitulate AATD liver disease.

FIG. 103 depicts current liver cell therapies have limitations.

FIG. 104 depicts proposed strategies to improve survival, proliferation, maturation, and engraftment of PHHs and HLCs in NSG-PiZ mice, which include stimulating key regenerative pathways in transplanted hepatocytes; preconditioning the host liver to prevent host hepatocyte proliferation; and maturing transplanted cells in vivo.

FIGS. 105 A- 105 D depict mRNA-LNP induces robust protein expression in the liver, mainly in hepatocytes.

FIGS. 106 A- 106 B depict HGF+EGF mRNA-LNP induces hepatocyte proliferation in vivo.

FIGS. 107 A- 107 D demonstrate HGF+EGF mRNA-LNP significantly increase PHH cluster size in NSG-PiZ mice following 2 weeks engraftment.

FIGS. 108 A- 108 C demonstrate HGF+EGF mRNA-LNP significantly increase PHH cluster size in NSG-PiZ mice after 9 weeks of engraftment.

FIGS. 109 A- 109 C demonstrate P21 expression delivered with AAV8-Tbg-p21 blocks host hepatocyte proliferation.

FIGS. 110 A- 110 D demonstrate AAV8-Tbg-P21 in addition to HGF+EGF mRNA-LNP significantly augments PHH repopulation in NSG-PiZ mice after 2 weeks engraftment.

FIGS. 111 A- 111 C demonstrate AAV8-Tbg-P21 and HGF+EGF mRNA-LNP drastically improve PHH engraftment after 5 week-engraftment.

FIGS. 112 A- 112 C demonstrate HGF+EGF mRNA-LNP transiently improves HCL survival after transplantation.

›DETAILED DESCRIPTION · 1 of 32

As described herein, the inventors have designed engineered liver regenerative factor compositions that provide improved therapeutic performance as compared to wild-type liver regenerative factors. These liver regenerative factor compositions can comprise one or more of: engineered mRNA sequences, engineered nucleosides (e.g., modified nucleosides); and carrier molecules and compositions.

As used herein, “liver regenerative factor” refers to a polypeptide that promotes or increases the growth, repair, function, or regeneration of liver tissue or cells, or a gene or mRNA encoding such a polypeptide. Such liver regenerative factors include but are not limited to: vascular endothelial growth factor A (VEGFA); hepatocyte growth factor (HGF); growth hormone (GH); insulin-like growth factor 1 (IGF-1), epidermal growth factor (EGF); signal transducer and activator of transcription 5B (STAT5b) (e.g., constitutively active STAT5b); cyclin-dependent kinase inhibitor 1A (p21); beta catenin (CTNNB1) (e.g., activated beta catenin); yes-associated protein (YAP) (e.g., activated YAP); wingless-type MMTV integration site family, member 2 (WNT2); and wingless-type MMTV integration site family, member 9B (WNT9b).

The sequences and structures of the foregoing liver regenerative factors are known in the art.

As used herein, “Growth Hormone” or “GH” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that, upon recognition by a receptor, stimulates MAPK/ERK and JAK-STAT signaling to increase IGF-1 production. The sequences of GH are known in the art for a number of species, e.g., human GH (NBCI Gene ID: 2688, polypeptide sequences NP_000506.2, NP_072053.1, NP_072054.1 and mRNA sequences NM_000515.5, NM_022559.4, and NM_022560.4) and murine GH (NBCI Gene ID: 14599, polypeptide sequence NP_032143.1 and mRNA sequence NM_008117.3). The structure and function of GH is known in the art. An exemplary wild-type mRNA sequence of GH is provided herein as SEQ ID NO: 1.

As used herein, “Epidermal Growth Factor” or “EGF” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that, upon recognition by the cognate receptor, stimulates cellular proliferation, differentiation, and survival and DNA synthesis. The sequences of EGF are known in the art for a number of species, e.g., human EGF (NBCI Gene ID: 1950, polypeptide sequences NP_001171601.1, NP_001171602.1, NP_001343950.1, and NP_001954.2 and mRNA sequences NM_001178130.3, NM_001178131.3, NM_001357021.2, and NM_001963.6) and murine EGF (NBCI Gene ID: 13645, polypeptide sequences NP_001297666.1, NP_001316523.1, and NP_034243.2 mRNA sequences NM_001310737.1, NM 001329594.1, and NM_010113.4). The structure and function of EGF is known in the art. An exemplary wild-type mRNA sequence of EGF is provided herein as SEQ ID NO: 2. As used herein, “secreted EGF” refers to a matured form of EGF in which the transmembrane region of EGF is cleaved. For example in NP_001954.2 the secreted form of EGF is provided in amino acids 971-1023.

As used herein, “Hepatocyte Growth Factor” or “HGF” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that, upon recognition by the c-Met receptor, stimulates mitogenesis, cell motility, and matrix invasion. HGF is a key factor in angiogenesis and tissue regeneration processes. The sequences of HGF are known in the art for a number of species, e.g., human HGF (NBCI Gene ID: 3082, polypeptide sequences NP_000592.3, NP_001010931.1, NP_001010932.1, NP_001010933.1, and NP_001010934.1 and mRNA sequences NM_000601.6, NM_001010931.3, NM_00101932.3, NM_001010933.3, and NM_001010934.3) and murine HGF (NBCI Gene ID: 15234, polypeptide sequences NP_001276387.1, NP_001276388.1, NP_001276389.1, NP_001276390.1, and NP_034557.3 and mRNA sequences NM_001289458.1, NM_001289459.1, NM_001289460.2, NM_001289461.1, and NM_010427.5). The structure and function of HGF is known in the art. An exemplary wild-type mRNA sequence of HGF is provided herein as SEQ ID NO: 3.

As used herein, “Cyclin-dependent Kinase inhibitor 1” or “p21” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that inhibits cyclin/CDK complexes, particularly CDK2. p21 also inhibits PCNA and apoptosis. The sequences of p21 are known in the art for a number of species, e.g., human p21 (NBCI Gene ID: 1026, polypeptide sequences NP_000380.1, NP_001207706.1, NP_001207707.1, NP_001278478.1, NP_001361438.1, NP 001361439.1, NP_001361440.1, NP_001361441.1, NP_001361442.1, and NP_510867.1 and mRNA sequences NM_000389.5, NM_001220777.2, NP_001220778.2, NM_001291549.3, NM_001374509.1, NM_001374510.1, NM_001374511.1, NM_001374512.1, NM_001374513.1, and NM_078467.3) and murine p21 (NBCI Gene ID: 12575, polypeptide sequences NP_001104569.1 and NP_031695.1 and mRNA sequences NM_001111099.2 and NM_007669.5). The structure and function of p21 is known in the art. An exemplary wild-type mRNA sequence of p21 is provided herein as SEQ ID NO: 4.

As used herein, “Vascular Endothelial Growth Factor” or “VEGF” or “VEGFA” refers to a cysteine-knot growth factor (or the gene or mRNA encoding said factor) that promotes vasculogenesis. A number of isoforms of VEGF are known, including VEGF 165 . The sequences of VEGF are known in the art for a number of species, e.g., human VEGF (NBCI Gene ID: 7422, polypeptide sequences NP 001020537.2, NP_001020538.2, NP_001020539.2, NP 001020540.2, NP_001020541.2, NP 001028928.1, NP_001165093.1, NP_001165094.1, NP_001165095.1, NP_001165096.1, NP_001165097.1, NP_001165098.1, NP_001165099.1, NP 001165100.1, NP_001165101.1, NP 001191313.1, NP_001191314.1, NP_001273973.1, NP_001303939.1, and NP_003367.4 and mRNA sequences NM_001025366.3, NM_00102367.3, NM_001025368.3, NM_001025369.3, NM 001025370.3, NM 001033756.3, NM_001171622.2, NM_001171623.2, NM_001171624.2, NM 001171625.2, NM 001171626.2, NM_001171627.2, NM_001171628.2, NM_001171629.2, NM 001171630.2, NM 001204384.2, NM_001204385.2, NM_001287044.2, NM_001317010.1, NM_003376.6) and murine VEGF (NBCI Gene ID: 22339, polypeptide sequences NP_001020421.2, NP 001020428.2, NP_001103736.1, NP 00103737.1, NP 001103738.1, NP_001273985.1, NP_001273986.1, NP_001273987.1, NP_001303970.1, and NP_033531.3 and mRNA sequences NM 001025250.3, NM 001025257.3, NM_001110266.1, NM_001110267.1, NM_001110268.1, NM_001287056.1, NM_001287057.1, NM_001287058.1, NM_001317041.1, and NM_009505.4). The sequence of VEGF165 is known in the art as well, e.g., human VEGF165 mRNA is provided in NCBI as AF486837.1. The structure and function of VEGF is known in the art. An exemplary wild-type mRNA sequence of VEGF is provided herein as SEQ ID NO: 5.

›DETAILED DESCRIPTION · 2 of 32

As used herein, “Insulin-Like Growth Factor 1” or “IGF-1” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) produced primarily in the liver in response to GH and which stimulates growth. The sequences of IGF-1 are known in the art for a number of species, e.g., human IGF-1 (NBCI Gene ID: 3479, polypeptide sequences NP_000609.1, NP_001104753.1, NP_001104754.1, and NP_001104755.1 and mRNA sequences NM_000618.5, NM_001111283.3, NM_001111284.2, and NM_001111285.3) and murine IGF-1 (polypeptide sequences NP_001104744.1, NP_001104745.1, NP_001104746.1, NP_001300939.1, and NP_034642.2 and mRNA sequences NM_001111274.1, NM_001111275.2, NM_001111276.1, NM_001314010.1, and NM_010512.5). The structure and function of IGF-1 is known in the art. An exemplary wild-type mRNA sequence of IGF-1 is provided herein as SEQ ID NO: 6.

As used herein, “Signal transducer and activator of transcription 5B” or “STAT5” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that is transcription factor responsive to IL2, IL4, CSF1, and growth factors. The sequences of STAT5B are known in the art for a number of species, e.g., human STAT5B (NBCI Gene ID: 6777, polypeptide sequences XP_047292549.1, XP_024306665.1, XP_016880466.1, XP_024306666.1, and XP_005257683.1 and mRNA sequences XM_047436593.1, XM_024450897.2, XM_017024977.2, XM_024450898.2, and XM_005257626.5) and murine STAT5B (NCBI Gene ID: 20851, polypeptide sequences NP_001107035.1, NP_001349611.1, and NP_035619.3 and mRNA sequences NM_001113563.2, NM_001362682.1, and NM_011489.3). The structure and function of STAT5B is known in the art. An exemplary wild-type mRNA sequence of STAT5B is provided herein as SEQ ID NO: 7.

As used herein, “beta catenin” or “β-catenin” or “CTNNB1” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that regulates cell to cell adhesion and gent transcription. The sequences of beta catenin are known in the art for a number of species, e.g., human beta catenin (NBCI Gene ID: 1499, polypeptide sequences NP_001091679.1, NP_001091680.1, NP_001317658.1, and NP_001895.1 and mRNA sequences NM_001098209.2, NM_001098210.2, NM_001330729.2, and NM_001904.4) and murine beta catenin (NCBI Gene ID: 12387, polypeptide sequences NP_001159374.1 and NP_031640.1 and mRNA sequences NM_001165902.1 and NM_007614.3). The structure and function of beta catenin is known in the art. An exemplary wild-type mRNA sequence of beta catenin is provided herein as SEQ ID NO: 17. In some embodiments of any of the aspects, beta catenin is activated beta-catenin.

As used herein, “Yes-associated protein 1” or “YAP” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that is a transcription co regulator of proliferation and apoptotic genes. The sequences of YAP are known in the art for a number of species, e.g., human YAP (NBCI Gene ID: 10413, polypeptide sequences NP_001123617.1, NP_001181973.1, NP_001181974.1, NP_001269026.1, NP_001269027.1, NP_001269028.1, NP_001269029.1, NP 001269030.1, NP_006097.2 and mRNA sequences NM_001130145.3, NM_001195044.2, NM_001195045.2, NM_001282097.2, NM_001282098.2, NM_001282099.2, NM_001282100.2, NM_001282101.2, and NM_006106.5) and murine YAP (NCBI Gene ID: 22601, polypeptide sequences NP_001164618.1 and NP_033560.1 and mRNA sequences NM_001171147.1 and NM_009534.3). The structure and function of YAP is known in the art. An exemplary wild-type mRNA sequence of YAP is provided herein as SEQ ID NO: 22. In some embodiments of any of the aspects, YAP is activated YAP.

As used herein, “Wingless-type MMTV integration site family, member 2” or “Wnt2” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that participates in the Wnt signaling pathway to regulate developmental and growth processes. The sequences of Wnt2 are known in the art for a number of species, e.g., human Wnt2 (NBCI Gene ID: 7472, polypeptide sequence NP_003382.1 and mRNA sequence NM_003391.3) and murine Wnt2 (NCBI Gene ID: 22413, polypeptide sequence NP_076142.3 and mRNA sequence NM_023653.5). The structure and function of Wnt2 is known in the art. An exemplary wild-type mRNA sequence of Wnt2 is provided herein as SEQ ID NO: 23.

As used herein, “Wingless-type MMTV integration site family, member 91B” or “Wnt9b” refers to a polypeptide (or the gene or mRNA encoding said polypeptide) that participates in the Wnt signaling pathway to regulate developmental and growth processes. The sequences of Wnt9b are known in the art for a number of species, e.g., human Wnt9b (NBCI Gene ID: 7484, polypeptide sequences NP_001307387.1 and NP_003387.1 and mRNA sequences NM_001320458.2 and NM_003396.3) and murine Wnt9b (NCBI Gene ID: 22412, polypeptide sequence NP_035849.3 and mRNA sequence NM_011719.4). The structure and function of Wnt9b is known in the art. An exemplary wild-type mRNA sequence of Wnt9b is provided herein as SEQ ID NO: 24.

Where reference is made herein to NCBI sequences and entries, it shall be understood that the NCBI sequences and data available under the provided numbers as of Nov. 8, 2022 are referred to.

In some embodiments of any of the aspects, the liver regenerative factor is or is derived from a mammalian liver regenerative factor. In some embodiments of any of the aspects, the liver regenerative factor is or is derived from a primate liver regenerative factor. In some embodiments of any of the aspects, the liver regenerative factor is or is derived from a human liver regenerative factor. In some embodiments of any of the aspects, the liver regenerative factor is or is derived from a murine liver regenerative factor. In some embodiments of any of the aspects, the liver regenerative factor is or is derived from a human liver regenerative factor or a murine liver regenerative factor.

In some embodiments of any of the aspects, a liver regenerative factor composition described herein relates to one or more mRNAs in which the mRNA sequence is not a naturally-occurring sequence, e.g., in which the mRNA sequence has been engineered to comprise one or more deletions, modifications, or additions. These engineered mRNA sequences can exhibit improve therapeutic characteristics including improved expression levels, cellular specificity, or half-life. Accordingly, in one aspect of any of the embodiments, described herein is a composition comprising at least one engineered liver regenerative factor mRNA, the at least one engineered liver regenerative factor mRNA comprising one or more of the deletions, modifications, and/or additions described herein.

›DETAILED DESCRIPTION · 3 of 32

In some embodiments of any of the aspects, the liver regenerative factor is GH and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 8. In some embodiments of any of the aspects, the liver regenerative factor is EGF and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 9. In some embodiments of any of the aspects, the liver regenerative factor is HGF and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 10. In some embodiments of any of the aspects, the liver regenerative factor is p21 and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 11. In some embodiments of any of the aspects, the liver regenerative factor is VEGF165 and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 12. In some embodiments of any of the aspects, the liver regenerative factor is IGF-1 and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 13. In some embodiments of any of the aspects, the liver regenerative factor is IGF-1 IL-2 SP and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 14. In some embodiments of any of the aspects, the liver regenerative factor is secreted EGF and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 15. In some embodiments of any of the aspects, the liver regenerative factor is STAT5B and the at least one engineered liver regenerative factor mRNA is constitutively activated and comprises SEQ ID NO: 16. In some embodiments of any of the aspects, the liver regenerative factor is beta-catenin and the at least one engineered liver regenerative factor mRNA is activated and comprises SEQ ID NO: 21. In some embodiments of any of the aspects, the liver regenerative factor is YAP and the at least one engineered liver regenerative factor mRNA is activated and comprises SEQ ID NO: 25. In some embodiments of any of the aspects, the liver regenerative factor is WNT2 and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 26. In some embodiments of any of the aspects, the liver regenerative factor is WNT9b and the at least one engineered liver regenerative factor mRNA comprises SEQ ID NO: 27.

In some embodiments of any of the aspects, the liver regenerative factor is GH and the at least one engineered liver regenerative factor mRNA consists of or consists essentially of SEQ ID NO: 8. In some embodiments of any of the aspects, the liver regenerative factor is EGF and the at least one engineered liver regenerative factor mRNA consists of or consists essentially of SEQ ID NO: 9. In some embodiments of any of the aspects, the liver regenerative factor is HGF and the at least one engineered liver regenerative factor mRNA consists of or consists essentially of SEQ ID NO: 10. In some embodiments of any of the aspects, the liver regenerative factor is p21 and the at least one engineered liver regenerative factor mRNA consists of or consists essentially of SEQ ID NO: 11. In some embodiments of any of the aspects, the liver regenerative factor is VEGF165 and the at least one engineered liver regenerative factor mRNA consists of or consists essentially of SEQ ID NO: 12. In some embodiments of any of the aspects, the liver regenerative factor is IGF-1 and the at least one engineered liver regenerative factor mRNA consists of or consists essentially of SEQ ID NO: 13. In some embodiments of any of the aspects, the liver regenerative factor is IGF-1 IL-2 SP and the at least one engineered liver regenerative factor mRNA consists of or consists essentially of SEQ ID NO: 14. In some embodiments of any of the aspects, the liver regenerative factor is secreted EGF and the at least one engineered liver regenerative factor mRNA consists of or consists essentially of SEQ ID NO: 15. In some embodiments of any of the aspects, the liver regenerative factor is STAT5B and the at least one engineered liver regenerative factor mRNA is constitutively active and consists of or consists essentially of SEQ ID NO: 16. In some embodiments of any of the aspects, the liver regenerative factor is beta-catenin and the at least one engineered liver regenerative factor mRNA is activated and consists of or consists essentially of SEQ ID NO: 21. In some embodiments of any of the aspects, the liver regenerative factor is YAP and the at least one engineered liver regenerative factor mRNA is activated and consists of or consists essentially SEQ ID NO: 25. In some embodiments of any of the aspects, the liver regenerative factor is WNT2 and the at least one engineered liver regenerative factor mRNA consists of or consists essentially SEQ ID NO: 26. In some embodiments of any of the aspects, the liver regenerative factor is WNT9b and the at least one engineered liver regenerative factor mRNA consists of or consists essentially SEQ ID NO: 27.

In some embodiments of any of the aspects, the liver regenerative factor is GH and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 8. In some embodiments of any of the aspects, the liver regenerative factor is EGF and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 9. In some embodiments of any of the aspects, the liver regenerative factor is HGF and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 10. In some embodiments of any of the aspects, the liver regenerative factor is p21 and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 11. In some embodiments of any of the aspects, the liver regenerative factor is VEGF165 and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 12. In some embodiments of any of the aspects, the liver regenerative factor is IGF-1 and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 13. In some embodiments of any of the aspects, the liver regenerative factor is IGF-1 IL-2 SP and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 14. In some embodiments of any of the aspects, the liver regenerative factor is EGF and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 15. In some embodiments of any of the aspects, the liver regenerative factor is STAT5B and the at least one engineered liver regenerative factor mRNA is constitutively active and comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 16. In some embodiments of any of the aspects, the liver regenerative factor is beta-catenin and the at least one engineered liver regenerative factor mRNA is activated and comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 21. In some embodiments of any of the aspects, the liver regenerative factor is YAP and the at least one engineered liver regenerative factor mRNA is activated and comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 25. In some embodiments of any of the aspects, the liver regenerative factor is WNT2 and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 26. In some embodiments of any of the aspects, the liver regenerative factor is WNT9b and the at least one engineered liver regenerative factor mRNA comprises, consists of, or consists essentially of a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or greater sequence identity to SEQ ID NO: 27.

›DETAILED DESCRIPTION · 4 of 32

In some embodiments of any of the aspects, the liver regenerative factor is GH and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 8. In some embodiments of any of the aspects, the liver regenerative factor is EGF and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 9. In some embodiments of any of the aspects, the liver regenerative factor is HGF and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 10. In some embodiments of any of the aspects, the liver regenerative factor is p21 and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 11. In some embodiments of any of the aspects, the liver regenerative factor is VEGF165 and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 12. In some embodiments of any of the aspects, the liver regenerative factor is IGF-1 and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% greater sequence identity to SEQ ID NO: 13. In some embodiments of any of the aspects, the liver regenerative factor is IGF-1 IL-2 SP and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 14. In some embodiments of any of the aspects, the liver regenerative factor is EGF and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 15. In some embodiments of any of the aspects, the liver regenerative factor is STAT5B and the at least one engineered liver regenerative factor mRNA is constitutively activated and comprises a sequence having at least 80% sequence identity to SEQ ID NO: 16. In some embodiments of any of the aspects, the liver regenerative factor is beta-catenin and the at least one engineered liver regenerative factor mRNA is activated and comprises a sequence having at least 80% sequence identity to SEQ ID NO: 21. In some embodiments of any of the aspects, the liver regenerative factor is YAP and the at least one engineered liver regenerative factor mRNA is activated and comprises a sequence having at least 80% sequence identity to SEQ ID NO: 25. In some embodiments of any of the aspects, the liver regenerative factor is WNT2 and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 26. In some embodiments of any of the aspects, the liver regenerative factor is WNT9b and the at least one engineered liver regenerative factor mRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 27.

In some embodiments of any of the aspects, the composition comprises a liver regenerative factor and at least one internal label or tag. In some embodiments, the tag can be, for example, a myc or FLAG tag. In some embodiments, the tag is a FLAG tag, for example, a FLAG tag encoded by the nucleic acid of SEQ ID NO: 28-29. In some embodiments the tag is a myc tag, for example, a myc tag encoded by the nucleic acid of SEQ ID NO: 30-31.

In some embodiments of any of the aspects, the at least one engineered liver regenerative factor is beta catenin and the at least one internal tag is a myc tag encoded by the nucleic acid of SEQ ID NO: 30 or SEQ ID NO: 31.

In some embodiments of any of the aspects, the tag is located after between sequence nucleotide 3 and nucleotide 4 of SEQ ID NO: 21. In some embodiments of any of the aspects, the tag is located after between sequence nucleotide 3 and nucleotide 4 of SEQ ID NO: 22. In some embodiments of any of the aspects, the tag is a myc tag is located between nucleotide 3 and nucleotide 4 of SEQ ID NO: 21. In some embodiments of any of the aspects, the tag is a FLAG tag is located between nucleotide 3 and nucleotide 4 of SEQ ID NO: 22.

In some embodiments of any of the aspects, a composition comprises two or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a composition comprises three or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a composition comprises four or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a composition comprises five or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a composition comprises six or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a composition comprises seven or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a composition comprises eight or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a composition comprises nine or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a composition comprises ten or more of the engineered liver regenerative factor mRNA sequences described herein.

In some embodiments of any of the aspects, a composition comprises two or more of SEQ ID NOs: 8-16, 21, and 25-27. In some embodiments of any of the aspects, a composition comprises three or more of SEQ ID NOs: 8-16, 21, and 25-27. In some embodiments of any of the aspects, a composition comprises four or more of SEQ ID NOs: 8-16, 21, and 25-27. In some embodiments of any of the aspects, a composition comprises five or more of SEQ ID NOs: 8-16, 21, and 25-27. In some embodiments of any of the aspects, a composition comprises six or more of SEQ ID NOs: 8-16, 21, and 25-27. In some embodiments of any of the aspects, a composition comprises seven or more of SEQ ID NOs: 8-16, 21, and 25-27. In some embodiments of any of the aspects, a composition comprises eight or more of SEQ ID NOs: 8-16, 21, and 25-27. In some embodiments of any of the aspects, a composition comprises nine or more of SEQ ID NOs: 8-16, 21, and 25-27. In some embodiments of any of the aspects, a composition comprises each of.

›DETAILED DESCRIPTION · 5 of 32

In some embodiments of any of the aspects, a mRNA, e.g., a mRNA molecule can be polycistronic, e.g., it can comprise two or more of the engineered liver regenerative factor mRNA sequences described herein. In some embodiments of any of the aspects, a mRNA, e.g., a mRNA molecule can be polycistronic, e.g., it can comprise two or more of SEQ ID NOs: 8-16, 21, and 25-27.

In embodiments comprising two or more liver regenerative factors, any combination of the liver regenerative factors and engineered liver regenerative factor mRNAs described herein is contemplated. Merely as illustrative examples, the following tables present exemplary pair-wise combinations of liver regenerative factors and engineered liver regenerative factor mRNAs.

In some embodiments of any of the aspects, a composition described herein can comprise two or more different engineered liver regenerative factor mRNAs of a single liver regenerative factor, e.g., two different GH engineered liver regenerative factor mRNAs.

In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of VEGFA; HGF; GH; and EGF. In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of: HGF; GH; EGF; and p21. In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of: HGF; GH; and EGF. In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of: VEGFA; HGF; GH; IGF-1; EGF; STAT5b; p21; CNNTB1; YAP; WNT2; and WNT9b.

In some embodiments of any of the aspects, the at least one liver regenerative factor mRNA (or engineered liver regenerative factor mRNA) is selected from the group consisting of: VEGFA mRNA; HGF mRNA; GH mRNA; and EGF mRNA. In some embodiments of any of the aspects, the at least one liver regenerative factor mRNA (or engineered liver regenerative factor mRNA) is selected from the group consisting of: HGF mRNA; GH mRNA; EGF mRNA; and p21 mRNA. In some embodiments of any of the aspects, the at least one liver regenerative factor mRNA (or engineered liver regenerative factor mRNA) is selected from the group consisting of: HGF mRNA; GH mRNA; and EGF mRNA. In some embodiments of any of the aspects, the at least one liver regenerative factor mRNA (or engineered liver regenerative factor mRNA) is selected from the group consisting of: VEGFA mRNA; HGF mRNA; GH mRNA; IGF-1 mRNA; EGF mRNA; STAT5b mRNA; p21 mRNA; CNNTB1 mRNA; YAP mRNA; WNT2 mRNA; and WNT9b mRNA.

In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of: VEGF165; HGF; GH; and EGF. In some embodiments of any of the aspects, the at least one liver regenerative factor is selected from the group consisting of: VEGF165; HGF; GH; IGF-1; EGF; STAT5b; p21; CNNTB1; YAP; WNT2; and WNT9b.

In some embodiments of any of the aspects, the at least one liver regenerative factor does not comprise VEGFA. In some embodiments of any of the aspects, the at least one liver regenerative factor does not comprise VEGF165.

As described herein, the nucleosides of a liver regenerative factor mRNA can also be engineered, e.g., modified to provide improved performance relative to wild-type mRNAs. In some embodiments of any of the aspects, an engineered liver regenerative factor mRNA can comprise at least one modified nucleoside. In one aspect of any of the embodiments, described herein is a composition comprising at least one liver regenerative factor mRNA (e.g., having a naturally-occurring mRNA sequence) comprising at least one modified nucleoside.

As used herein, “modified nucleoside” refers to a nucleoside that is not naturally-occurring, e.g., a not naturally-occurring sugar and/or nucleobase. Exemplary modified sugars can include, e.g., one of the following at the 2′ position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C1 to C10 alkyl or C2 to C10 alkenyl and alkynyl. Exemplary suitable modifications include O[(CH2)nO] mCH3, O(CH2)·nOCH3, O(CH2)nNH2, O(CH2)nCH3, O(CH2)nONH2, and O(CH2)nON[(CH2)nCH3)]2, where n and m are from 1 to about 10. In some embodiments of any of the aspects, a modified nucleoside can include one of the following at the 2′ position: C1 to C10 lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ONO2, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an mRNA, or a group for improving the pharmacodynamic properties of an mRNA, and other substituents having similar properties. In some embodiments of any of the aspects, the modified nucleoside includes a 2′ methoxyethoxy (2′-O—CH2CH2OCH3, also known as 2′-O-(2-methoxyethyl) or 2′-MOE) (Martin et al., Helv. Chim. Acta, 1995, 78:486-504) i.e., an alkoxy-alkoxy group. Another exemplary modification is 2′-dimethylaminooxyethoxy, i.e., a O(CH2)2ON(CH3)2 group, also known as 2′-DMAOE and 2′-dimethylaminoethoxyethoxy (also known in the art as 2′-O-dimethylaminoethoxyethyl or 2′-DMAEOE), i.e., 2′-O—CH2-O—CH2-N(CH2)2.

Other modifications include 2′-methoxy (2′-OCH3), 2′-aminopropoxy (2′-OCH2CH2CH2NH2) and 2′-fluoro (2′-F). Modified nucleosides may also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar.

›DETAILED DESCRIPTION · 6 of 32

A modified nucleoside can also include nucleobase (often referred to in the art simply as “base”) modifications or substitutions. As used herein, “unmodified” or “natural” nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U). Modified nucleobases include other synthetic and natural nucleobases such as 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl anal other 8-substituted adenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.

In some embodiments of any of the aspects, a modified nucleoside can be pseudouridine, N1-methylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine, pseudouridine, 5-methyluridine, 5-methoxyuridine, or 2′-O-methyluridine.

In some embodiments of any of the aspects, a modified nucleoside can be 2′-O-Methylpseudouridine-5′-Triphosphate; N1-Methylpseudouridine-5′-Triphosphate; N1-Methyl-2′-O-Methylpseudouridine-5′-Triphosphate; or 1-Methylpseudouridine. In some embodiments of any of the aspects, a modified nucleoside can be one-methylpseudouridine (m1Ψ)-5′ triphosphate.

In some embodiments of any of the aspects, a modified nucleoside is a non-natural nucleoside. In some embodiments of any of the aspects, an engineered liver regenerative factor mRNA or liver regenerative factor mRNA described herein does not comprise a modified nucleoside, e.g., it comprises only naturally-occurring nucleosides.

In some embodiments of any of the aspects, a mRNA comprising at least one modified nucleoside comprises at least one one-methylpseudouridine (m1Ψ)-5′-triphosphate residue.

In some embodiments of any of the aspects, a mRNA comprises, relative to a reference sequence provided herein, the replacement of at least one uracil or thymine with a modified nucleoside. In some embodiments of any of the aspects, a mRNA comprises, relative to a reference sequence provided herein, the replacement of at least one uracil or thymine with one-methylpseudouridine (m1Ψ)-5′-triphosphate.

In some embodiments of any of the aspects, a mRNA comprises, relative to a reference sequence provided herein, the replacement of at least half of the uracils or thymines with modified nucleosides. In some embodiments of any of the aspects, a mRNA comprises, relative to a reference sequence provided herein, the replacement of at least half of the uracils or thymines with one-methylpseudouridine (m1Ψ)-5′-triphosphates.

In some embodiments of any of the aspects, a mRNA comprises, relative to a reference sequence provided herein, the replacement of each uracil or thymine with a modified nucleoside. In some embodiments of any of the aspects, a mRNA comprises, relative to a reference sequence provided herein, the replacement of each uracil or thymine with one-methylpseudouridine (m1Ψ)-5′-triphosphate.

In some embodiments of any of the aspects, a mRNA comprises a m7G cap.

The preparation of the modified nucleosides described above are well known in the art.

In some embodiments of any of the aspects, contacting a cell or administering to a subject one of the compositions or combinations described herein, e.g., comprising an engineered liver regenerative factor mRNA and/or liver regenerative factor mRNA comprising at least one modified nucleoside increases the protein level of the liver regenerative factor in the cell/subject. In some embodiments of any of the aspects, contacting a cell or administering to a subject one of the compositions or combinations described herein, e.g., comprising an engineered liver regenerative factor mRNA and/or liver regenerative factor mRNA comprising at least one modified nucleoside increases the protein level of the liver regenerative factor in the liver of the subject. In some embodiments of any of the aspects, the increase is relative to the absence of, or prior to, the contacting or administering. Methods of measuring the protein level of a factor are well known in the art and include, e.g, immunochemistry, immunohistochemistry (“IHC”), immunocytochemistry (“ICC”), Western blot, ELISA, and LFIA methods or the like.

In some embodiments of any of the aspects, the composition further comprises a carrier or scaffold complexed with the at least one engineered liver regenerative factor mRNA (comprising or not comprising at least one modified nucleoside) and/or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside. Carrier or scaffold materials suitable for use in therapeutic compositions are known in the art and can include, but are not limited to, a nanoparticle; a matrix; a hydrogel; and a biomaterial, biocompatible, and/or biodegradable scaffold material.

In some embodiments of any of the aspects, the carrier is, comprises, or consists of a nanoparticle. As used herein, the term “nanoparticle” refers to particles that are on the order of about 10-9 or one to several billionths of a meter. The term “nanoparticle” includes nanospheres; nanorods; nanoshells; and nanoprisms; these nanoparticles may be part of a nanonetwork.

As used herein, the term “matrix” refers to a 3-dimensional structure comprising the components of a composition described herein (e.g., mRNA). Non-limiting examples of matrix structures include foams; hydrogels; electrospun fibers; gels; fiber mats; sponges; 3-dimensional scaffolds; non-woven mats; woven materials; knit materials; fiber bundles; and fibers and other material formats (See, e.g., Rockwood et al. Nature Protocols 2011 6:1612-1631 and US Patent Publications 2011/0167602; 2011/0009960; 2012/0296352; and U.S. Pat. No. 8,172,901; each of which is incorporated by reference herein in its entirety). The structure of the matrix can be selected by one of skill in the art depending upon the intended application of the composition, e.g., electrospun matrices can have greater surface area than foams.

›DETAILED DESCRIPTION · 7 of 32

In some embodiments, the carrier is a hydrogel. As used herein, the term “hydrogel” refers to a three-dimensional polymeric structure that is insoluble in water but which is capable of absorbing and retaining large quantities of water to form a stable, often soft and pliable, structure. In some embodiments, water can penetrate in between the polymer chains of the polymer network, subsequently causing swelling and the formation of a hydrogel. In general, hydrogels are superabsorbent. Hydrogels have many desirable properties for biomedical applications. For example, they can be made nontoxic and compatible with tissue, and they are highly permeable to water, ions, and small molecules. Hydrogels are super-absorbent (they can contain over 99% water) and can be comprised of natural (e.g., silk) or synthetic polymers, e.g., PEG.

As used herein, “biomaterial” refers to a material that is biocompatible and biodegradable. As used herein, the term “biocompatible” refers to substances that are not toxic to cells. In some embodiments, a substance is considered to be “biocompatible” if its addition to cells in vitro results in less than or equal to approximately 20% cell death. In some embodiments, a substance is considered to be “biocompatible” if its addition to cells in vivo does not induce inflammation and/or other adverse effects in vivo. As used herein, the term “biodegradable” refers to substances that are degraded under physiological conditions. In some embodiments, a biodegradable substance is a substance that is broken down by cellular machinery. In some embodiments, a biodegradable substance is a substance that is broken down by chemical processes.

The term “nanoparticles” also encompasses liposomes and lipid particles having the size of a nanoparticle. Exemplary liposomes can comprise, e.g., DSPC, DPPC, DSPG, Cholesterol, hydrogenated soy phosphatidylcholine, soy phosphatidyl choline, methoxypolyethylene glycol (mPEG-DSPE) phosphatidyl choline (PC), phosphatidyl glycerol (PG), distearoylphosphatidylcholine, and combinations thereof. In some embodiments of any of the aspects, the carrier is, comprises, or consists of a lipid nanoparticle (LNP). Lipid nanoparticles can comprise multiple components, including, e.g., ionizable lipids (such as MC3, DLin-MC3-DMA, ALC-0315, or SM-102), pegylated lipids (such as PEG2000-C-DMG, PEG2000-DMG, ALC-0159), phospholipids (such as DSPC), and cholesterol.

Generally, the lipid nanoparticles have a mean diameter selected to provide an intended therapeutic effect. Accordingly, in some aspects, the lipid nanoparticle has a mean diameter from about 30 nm to about 150 nm, more typically from about 50 nm to about 150 nm, more typically about 60 nm to about 130 nm, more typically about 70 nm to about 110 nm, most typically about 85 nm to about 105 nm, and preferably about 100 nm. In some aspects, the disclosure provides for lipid particles that are larger in relative size to common nanoparticles and about 150 to 250 nm in size. Lipid nanoparticle particle size can be determined by quasi-elastic light scattering using, for example, a Malvern Zetasizer Nano ZS (Malvern, UK) system.

Depending on the intended use of the lipid particles, the proportions of the components can be vaned and the delivery efficiency of a particular formulation can be measured using, for example, an endosomal release parameter (ERP) assay.

The mRNA can be complexed with the lipid portion of the particle or encapsulated in the lipid position of the lipid nanoparticle. In some embodiments, the mRNA can be fully encapsulated in the lipid position of the lipid nanoparticle, thereby protecting it from degradation by a nuclease, e.g., in an aqueous solution. In some embodiments, the mRNA in the lipid nanoparticle is not substantially degraded after exposure of the lipid nanoparticle to a nuclease at 37° C. for at least about 20, 30, 45, or 60 minutes. In some embodiments, the mRNA in the lipid nanoparticle is not substantially degraded after incubation of the particle in serum at 37° C. for at least about 30, 45, or 60 minutes or at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36 hours.

In certain embodiments, the lipid nanoparticles are substantially non-toxic to mammals such as humans.

In some embodiments, lipid nanoparticles are solid core particles that possess at least one lipid bilayer. In other embodiments, the lipid nanoparticles have a non-bilayer structure, i.e., a non-lamellar (i.e., non-bilayer) morphology. Without limitations, the non-bilayer morphology can include, for example, three dimensional tubes, rods, cubic symmetries, etc. The non-lamellar morphology (i.e., non-bilayer structure) of the lipid particles can be determined using analytical techniques known to and used by those of skill in the art. Such techniques include, but are not limited to, Cryo-Transmission Electron Microscopy (“Cryo-TEM”), Differential Scanning calorimetry (“DSC”), X-Ray Diffraction, etc. . . . . For example, the morphology of the lipid nanoparticles (lamellar vs. non-lamellar) can readily be assessed and characterized using, e.g., Cryo-TEM analysis as described in US2010/0130588, content of which is incorporated herein by reference in its entirety.

In some further embodiments, the lipid nanoparticles having a non-lamellar morphology are electron dense.

In embodiments, the lipid nanoparticle is either unilamellar or multilamellar in structure. In some aspects, the disclosure provides for a lipid nanoparticle formulation that comprises multi-vesicular particles and/or foam-based particles.

The lipid nanoparticle may have positive or negative zeta potential. In various embodiments, the lipid nanoparticle has a positive zeta potential.

By controlling the composition and concentration of the lipid components, one can control the rate at which the lipid conjugate exchanges out of the lipid particle and, in turn, the rate at which the lipid nanoparticle becomes fusogenic. In addition, other variables including, e.g., pH, temperature, or ionic strength, can be used to vary and/or control the rate at which the lipid nanoparticle becomes fusogenic. Other methods which can be used to control the rate at which the lipid nanoparticle becomes fusogenic will become apparent to those of skill in the art upon reading this disclosure. Also, by controlling the composition and concentration of the lipid conjugate, one can control the lipid particle size.

›DETAILED DESCRIPTION · 8 of 32

The pKa of formulated cationic lipids can be correlated with the effectiveness of the LNPs for delivery of nucleic acids (see Jayaraman et al, Angewandte Chemie, International Edition (2012), 51(34), 8529-8533; Semple et al, Nature Biotechnology 28, 172-176 (2010), both of which are incorporated by reference in their entirety). The preferred range of pKa is ˜5 to ˜7. The pKa of the cationic lipid can be determined in lipid nanoparticles using an assay based on fluorescence of 2-(p-toluidino)-6-napthalene sulfonic acid (TNS).

Encapsulation of mRNA in lipid particles can be determined by performing a membrane-impermeable fluorescent dye exclusion assay, which uses a dye that has enhanced fluorescence when associated with nucleic acid, for example, an Oligreen® assay or PicoGreen® assay. Generally, encapsulation is determined by adding the dye to the lipid particle formulation, measuring the resulting fluorescence, and comparing it to the fluorescence observed upon addition of a small amount of nonionic detergent. Detergent-mediated disruption of the lipid bilayer releases the encapsulated mRNA, allowing it to interact with the membrane-impermeable dye. Encapsulation of mRNA can be calculated as E=(I 0 −I)/I 0 , where I and I 0 refers to the fluorescence intensities before and after the addition of detergent.

Lipid nanoparticles can form spontaneously upon mixing of mRNA and the lipid(s). Depending on the desired particle size distribution, the resultant nanoparticle mixture can be extruded through a membrane (e.g., 100 nm cut-off) using, for example, a thermobarrel extruder, such as Lipex Extruder (Northern Lipids, Inc). In some cases, the extrusion step can be omitted. Ethanol removal and simultaneous buffer exchange can be accomplished by, for example, dialysis or tangential flow filtration.

Generally, lipid nanoparticles can be formed by any method known in the art including. For example, the lipid nanoparticles can be prepared by the methods described, for example, in US2013/0037977, US2010/0015218, US2013/0156845, US2013/0164400, US2012/0225129, and US2010/0130588, content of each of which is incorporated herein by reference in its entirety. In some embodiments, lipid nanoparticles can be prepared using a continuous mixing method, a direct dilution process, or an in-line dilution process. The processes and apparatuses for apparatuses for preparing lipid nanoparticles using direct dilution and in-line dilution processes are described in US2007/0042031, content of which is incorporated herein reference in its entirety. The processes and apparatuses for preparing lipid nanoparticles using step-wise dilution processes are described in US2004/0142025, content of which is incorporated herein reference in its entirety.

In one non-limiting example, the lipid nanoparticles can be prepared by an impinging jet process. Generally, the particles are formed by mixing lipids dissolved in alcohol (e.g., ethanol) with mRNA dissolved in a buffer, e.g., a citrate buffer, a sodium acetate buffer, a sodium acetate and magnesium chloride buffer, a malic acid buffer, a malic acid and sodium chloride buffer, or a sodium citrate and sodium chloride buffer. The mixing ratio of lipids to mRNA can be about 45-55% lipid and about 65-45% mRNA.

The lipid solution can contain a condensing agent (e.g., a cationic lipid), a non-cationic lipid (e.g., a phospholipid, such as DSPC), PEG or PEG conjugated molecule (e.g., PEG-lipid), and a sterol (e.g., cholesterol) at a total lipid concentration of 5-30 mg/mL, more likely 5-15 mg/mL, most likely 9-12 mg/mL in an alcohol, e.g., in ethanol.

In the lipid solution, mol ratio of the lipids can range from about 25-98% for the cationic lipid, preferably about 35-65%; about 0-15% for the non-ionic lipid, preferably about 0-12%; about 0-15% for the PEG or PEG conjugated molecule, preferably about 1-6%; and about 0-75% for the sterol, preferably about 30-50%.

The mRNA solution can comprise the mRNA at a concentration range from 0.3 to 1.0 mg/mL, preferably 0.3-0.9 mg/mL in buffered solution, with pH in the range of 3.5-5.

For forming the LNPs, the two liquids are heated to a temperature in the range of about 15-40° C., preferably about 30-40° C., and then mixed, for example, in an impinging jet mixer, instantly forming the LNP. The mixing flow rate can range from 10-600 ml/min. The tube ID can have a range from 0.25 to 1.0 mm and a total flow rate from 10-600 mL/min. The combination of flow rate and tubing ID can have the effect of controlling the particle size of the LNPs between 30 and 200 nm. The solution can then be mixed with a buffered solution at a higher pH with a mixing ratio in the range of 1:1 to 1:3 vol:vol, preferably about 1:2 vol:vol. If needed this buffered solution can be at a temperature in the range of 15-40° C. or 30-40° C. The mixed LNPs can then undergo an anion exchange filtration step. Prior to the anion exchange, the mixed LNPs can be incubated for a period of time, for example 30 mins to 2 hours. The temperature during incubating can be in the range of 15-40° C. or 30-40° C. After incubating the solution is filtered through a filter, such as a 0.8 μm filter, containing an anion exchange separation step. This process can use tubing IDs ranging from 1 mm ID to 5 mm ID and a flow rate from 10 to 2000 mL/min.

After formation, the LNPs can be concentrated and diafiltered via an ultrafiltration process where the alcohol is removed and the buffer is exchanged for the final buffer solution, for example, phosphate buffered saline (PBS) at about pH 7, e.g., about pH 6.9, about pH 7.0, about pH 7.1, about pH 7.2, about pH 7.3, or about pH 7.4.

The ultrafiltration process can use a tangential flow filtration format (TFF) using a membrane nominal molecular weight cutoff range from 30-500 KD. The membrane format is hollow fiber or flat sheet cassette. The TFF processes with the proper molecular weight cutoff can retain the LNP in the retentate and the filtrate or permeate contains the alcohol; citrate buffer and final buffer wastes. The TFF process is a multiple step process with an initial concentration to a mRNA concentration of 1-3 mg/mL. Following concentration, the LNPs solution is diafiltered against the final buffer for 10-20 volumes to remove the alcohol and perform buffer exchange. The material can then be concentrated an additional 1-3 fold. The concentrated LNP solution can be sterile filtered.

›DETAILED DESCRIPTION · 9 of 32

In some embodiments of any of the aspects, the compositions comprising mRNA as described herein are formulated in a lipid vesicle that have crosslinks between functionalized lipid bilayers.

In some embodiments, the compositions comprising mRNA(s) as described herein are formulated in a lipid-polycation complex. The formation of the lipid-polycation complex can be accomplished by methods known in the art and/or as described in U.S. Pub. No. 20120178702, herein incorporated by reference in its entirety. As a non-limiting example, the polycation can include a cationic peptide or a polypeptide such as, but not limited to, polylysine, polyornithine and/or polyarginine. In another embodiment, the polynucleotides, primary constructs and/or mRNA can be formulated in a lipid-polycation complex which can further include a neutral lipid such as, but not limited to, cholesterol or dioleoyl phosphatidylethanolamine (DOPE).

The selection of the liposome formulation can be influenced by, for example, the selection of the cationic lipid component, the degree of cationic lipid saturation, the nature of the PEGylation, ratio of all components and biophysical parameters such as size. In some embodiments, the ratio of PEG in the LNP formulations can be increased or decreased and/or the carbon chain length of the PEG lipid can be modified from C14 to C18 to alter the pharmacokinetics and/or biodistribution of the LNP formulations. The cationic lipid can be selected from any lipid known in the art such as, but not limited to, DLin-MC3-DMA, DLin-DMA, C12-200 and DLin-KC2-DMA.

In some embodiments, the cationic lipid can be selected from, but not limited to, a cationic lipid described in International Publication Nos. WO2012040184, WO2011153120, WO2011149733, WO2011090965, WO2011043913, WO2011022460, WO2012061259, WO2012054365, WO2012044638, WO2010080724, WO201021865 and WO2008103276, U.S. Pat. Nos. 7,893,302 and 7,404,969 and US Patent Publication No. US20100036115; each of which is herein incorporated by reference in their entireties. In another embodiment, the cationic lipid is selected from formula A described in International Publication Nos. WO2012040184, WO2011153120, WO2011149733, WO2011090965, WO2011043913, WO2011022460, WO2012061259, WO2012054365 and WO2012044638; each of which is herein incorporated by reference in their entireties. In other embodiments, the cationic lipid is selected from, formula CLI-CLXXIX of International Publication No. WO2008103276, formula CLI-CLXXIX of U.S. Pat. No. 7,893,302, formula CLI-CLXXXXII of U.S. Pat. No. 7,404,969 and formula I-VI of US Patent Publication No. 0520100036115; each of which is herein incorporated by reference in their entireties. As non-limiting examples, the cationic lipid is selected from (20Z,23Z)—N,N-dimethylnonacosa-20,23-dien-10-amine, (17Z,20Z)—N,N-dimemylhexacosa-17,20-dien-9-amine, (1Z,19Z)—N5N.about.dimethylpentacosa.about.16,19-dien-8-amine, (13Z,16Z)—N,N-dimethyldocosa-13J16-dien-5-amine, (12Z,15Z)—NJN-dimethylhenicosa-12,15-dien-4-amine, (14Z,17Z)—N,N-dimethyltricosa-14,17-dien-6-amine, (15Z,18Z)—N,N-dimethyltetracosa-15,18-dien-7-amine, (18Z,21Z)—N,N-dimethylheptacosa-18,21-dien-10-amine, (15Z,18Z)—N,N-dimethyltetracosa-15,18-dien-5-amine, (14Z,17Z)—N,N-dimethyltricosa-14,17-dien-4-amine, (19Z,22Z)—N,N-dimeihyloctacosa-19,22-dien-9-amine, (18Z,21Z)—N,N-dimethylheptacosa-18,21-dien-8-amine, (17Z,20Z)—N,N-dimethylhexacosa-17,20-dien-7-amine, (16Z;19Z)—N,N-dimethylpentacosa-16,19-dien-6-amine, (22Z,25Z)—N,N-dimethylhentriaconta-22,25-dien-10-amine, (21Z,24Z)—N;N-dimethyltriaconta-21,24-dien-9-amine, (18Z)—N,N-dimetylheptacos-18-en-10-amine, (17Z)—N,N-dimethylhexacos-17-en-9-amine, (19Z,22Z)—NJN-dimethyloctacosa-19,22-dien-7-amine, N,N-dimethylheptacosan-10-amine, (20Z,23Z)—N-ethyl-N-methylnonacosa-20J23-dien-10-amine, 1-[(11Z,14Z)-1-nonylicosa-11,14-dien-1-yl] pyrrolidine, (20Z)—N,N-dimethylheptacos-20-en-10-amine, (15Z)—N,N-dimethyl eptacos-15-en-10-amine, (14Z)—N,N-dimethylnonacos-14-en-10-amine, (17Z)—N,N-dimethylnonacos-17-en-10-amine, (24Z)—N,N-dimethyltritriacont-24-en-10-amine, (20Z)—N,N-dimethylnonacos-20-en-10-amine, (22Z)—N,N-dimethylhentriacont-22-en-10-amine, (16Z)—N,N-dimethylpentacos-16-en-8-amine, (12Z,15Z)—N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine, (13Z,16Z)—N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine, N,N-dimethyl-1-[(1 S,2R)-2-octylcyclopropyl] eptadecan-8-amine, 1-[(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecan-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]nonadecan-10-amine, N,N-dimethyl-21.about.[(1S,2R)-2-octylcyclopropyl]henicosan-1 O-amine, N,N-dimethyl-1-[(1 S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl} cyclopropyl]nonadecan-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]hexadecan-8-amine, N,N-dimethyH-[(1R,2S)-2-undecylcyclopropyl]tetradecan-5-amine, N,N-dimethyl-3-{7-[(1S,2R)-2-octylcyclopropyl]heptyl} dodec-an-1-amine, 1-[(1R,2S)-2-heptylcyclopropy 1]-N,N-dimethyloctadecan-9-amine, 1-[(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecan-6-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]pentadecan-8-amine, R—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octyloxy)propa-n-2-amine, S—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octyloxy)propan-2-amine, 1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-[(octyloxy) methyl]ethyl}pyrrolidine, (2S)—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-[(5Z)-oct-5-en-1-yloxy]propan-2-amine, 1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-[(octyloxy) methyl]ethyl}azetidine, (2S)-1-(hexyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]pro-pan-2-amine, (2S)-1-(heptyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]pr-opan-2-amine, N,N-dimethyl-1-(nonyloxy)-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, N,N-dimethyl-1-[(9Z)-octadec-9-en-1-yloxy]-3-(octyloxy)propan-2-am-me (Compound 9); (2S)—N,N-dimethyl-1-[(6Z,9Z,12Z)-octadeca-6,9,12-trien-1-yloxy]-3-(octyl-oxy)propan-2-amine, (2S)-1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(pentyloxy)pro-pan-2-amine, (2S)-1-(hexyloxy)-3-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethylprop-an-2-amine, 1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, 1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)pr-opan-2-amine, (2S)-1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-3-(hexyloxy)-N,N-dimethylpro-pan-2-amine, (2S)-1-[(13Z)-docos-13-en-1-yloxy]-3-(hexyloxy)-N,N-dimethylpropan-2-amin-e, 1-[(13Z)-docos-13-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, 1-[(9Z)-hexadec-9-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, (2R)—N,N-dimethyl-H(1-metoyloctyl)oxy]-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, (2R)-1-[(3,7-dimethyloctyl)oxy]-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-di-en-1-yloxy]propan-2-amine, N,N-dimethyl-1-(octyloxy)-3-({8-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]-methyl}cyclopropyl]octyl}oxy)propan-2-amine, N,N-dimethyl-1-{[8-(2-oclylcyclopropyl)octyl]oxy}-3-(octyloxy)propan-2-am-ine and (11E,20Z,23Z)—N;N-dimethylnonacosa-11,20,2-trien-10-amine or a pharmaceutically acceptable salt or stereoisomer thereof.

›DETAILED DESCRIPTION · 10 of 32

In some embodiments, the LNP formulations of the compositions comprising mRNA(s) as described herein can contain PEG-c-DOMG between and including 1.5-3% lipid molar ratio. In some embodiments, the compositions comprising the mRNA(s) described herein include at least one of the PEGylated lipids described in International Publication No. 2012099755, herein incorporated by reference. In some embodiments, the LNP formulation contains PEG-DMG 2000 (1,2-dimyristoyl-sn-glycero-3-phophoethanolamine-N-[methoxy(polyethylene glycol)-2000), a cationic lipid known in the art, and at least one other component. As a non-limiting example, the LNP formulation can contain PEG-DMG 2000, DLin-DMA, DSPC and cholesterol.

In some embodiments, the LNP formulation can be formulated by the methods described in International Publication Nos. WO2011127255 or WO2008103276, each of which is herein incorporated by reference in their entireties. As a non-limiting example, modified RNA can be encapsulated in LNP formulations as described in WO2011127255 and/or WO2008103276; each of which is herein incorporated by reference in their entireties.

In some embodiments, the LNP formulations comprise a polycationic composition. In some embodiments, the LNP formulations additionally comprise a permeability enhancer molecule. Non-limiting permeability enhancer molecules are described in US Patent Publication No. US20050222064; herein incorporated by reference in its entirety.

In some embodiments of any of the aspects, lipid nanoparticle formulations for use with compositions comprising mRNA(s) as described herein can be improved by replacing the cationic lipid with a biodegradable cationic lipid, which is known as a rapidly eliminated lipid nanoparticle (reLNP). The rapid metabolism of the rapidly eliminated lipids can improve the tolerability and therapeutic index of the lipid nanoparticles. Inclusion of an enzymatically degraded ester linkage can improve the degradation and metabolism profile of the cationic component, while still maintaining the activity of the reLNP formulation. The ester linkage can be internally located within the lipid chain or it can be terminally located at the terminal end of the lipid chain. The internal ester linkage can replace any carbon in the lipid chain. The internal ester linkage can be located on either side of the saturated carbon.

In some embodiments, compositions comprising mRNA(s) are formulated as a lipoplex, such as, without limitation, the ATUPLEX system, the DACC system, the DBTC system and other siRNA-lipoplex technology from Silence Therapeutics (London, United Kingdom), STEMFECT from STEMGENT (Cambridge, Mass.), and polyethylenimine (PEI) or protamine-based targeted and non-targeted delivery of nucleic acids (Aleku et al. Cancer Res. 2008 68:9788-9798; Strumberg et al. Int J Clin Pharmacol Ther 2012 50:76-78; Santel et al., Gene Ther 2006 13:1222-1234; Santel et al., Gene Ther 2006 13:1360-1370; Gutbier et al., Pulm Pharmacol. Ther. 2010 23:334-344; Kaufmann et al. Microvasc Res 2010 80:286-293 Weide et al. J Immunother. 2009 32:498-507; Weide et al. J Immunother. 2008 31:180-188; Pascolo Expert Opin. Biol. Ther. 4:1285-1294; Fotin-Mleczek et al., 2011 J. Immu-nother. 34:1-15; Song et al., Nature Biotechnol. 2005, 23:709-717; Peer et al., Proc Natl Acad Sci USA. 2007 6; 104:4095-4100; deFougerolles Hum Gene Ther. 2008 19:125-132; all of which are incorporated herein by reference in their entireties).

In some embodiments, the compositions comprising an mRNA(s) as described herein are formulated as a solid lipid nanoparticle. A solid lipid nanoparticle (SLN) can be spherical with an average diameter be-tween 10 to 1000 nm. SLN possess a solid lipid core matrix that can solubilize lipophilic molecules and can be stabilized with surfactants and/or emulsifiers. The lipid nanoparticle can be a self-assembly lipid-polymer nanoparticle (see Zhang et al., ACS Nano, 2008, 2 (8), pp 1696-1702; herein incorporated by reference in its entirety).

Liposomes, lipoplexes, or lipid nanoparticles can be used to improve the efficacy of protein production as these formulations can increase cell transfection by the compositions comprising mRNA(s) as described herein; and/or increase the translation of encoded protein. One such example involves the use of lipid encapsulation to enable the effective systemic delivery of polyplex plasmid DNA (Heyes et al., Mol Ther. 2007 15:713-720; herein incorporated by reference in its entirety). The liposomes, lipoplexes, or lipid nanoparticles can also be used to increase the stability of the compositions comprising mRNA(s) as described herein.

The compositions comprising RNA(s) as described herein can, in some embodiments, be encapsulated into a lipid nanoparticle or a rapidly eliminating lipid nanoparticle, and the lipid nanoparticles or a rapidly eliminating lipid nanoparticle may then be encapsulated into a polymer, hydrogel and/or surgical seal-ant described herein and/or known in the art. As a non-limiting example, the polymer, hydrogel or surgical sealant may be PLGA, ethylene vinyl acetate (EVAc), poloxamer, GELSITE (Nanotherapeutics, Inc. Alachua, Fla.), HYLENEX (Halozyme Therapeutics, San Diego Calif.), surgical sealants such as fibrinogen polymers (Ethicon Inc. Cornelia, Ga.), TISSELL (Baxter International, Inc Deerfield, Ill.), PEG-based sealants, and COSEAL. (Baxter International, Inc Deerfield, Ill.).

Compositions comprising mRNA(s) as described herein can include formulations in which the mRNA(s) is complexed or associated with one or more cationic polymers, including but not limited to polyethyleneimine and cationic dendrimers.

The compositions comprising mRNA(s) as described herein can be encapsulated in a therapeutic nanoparticle. Therapeutic nanoparticles can be formulated by methods described herein and known in the art such as, but not limited to, International Pub Nos. WO2010005740, WO2010030763, WO2010005721, WO2010005723, WO2012054923, US Pub. Nos. US20110262491, US20100104645, US20100087337, US20100068285, US20110274759, US20100068286, and U.S. Pat. No. 8,206,747; the contents of each of which are herein incorporated by reference in their entireties.

›DETAILED DESCRIPTION · 11 of 32

The nanoparticles used with the compositions comprising mRNA(s) as described herein can comprise a polymeric matrix. As a non-limiting example, the nanoparticle can comprise two or more polymers such as, but not limited to, polyethylenes, polycarbonates, polyanhydrides, polyhydroxyacids, polypropylfumerates, polycaprolactones, polyamides, polyacetals, polyethers, polyesters, poly(orthoesters), pol-ycyanoacrylates, polyvinyl alcohols, polyurethanes, polyphosphazenes, polyacrylates, polymethacry-lates, polycyanoacrylates, polyureas, polystyrenes, polyamines, polylysine, poly(ethylene imine), poly(serine ester), poly(L-lactide-co-L-lysine), poly(4-hydroxy-L-proline ester) or combinations thereof.

The nanoparticles used with the compositions comprising mRNA(s) as described herein can comprise at least one acrylic polymer. Acrylic polymers include but are not limited to, acrylic acid, methacrylic acid, acrylic acid and methacrylic acid copolymers, methyl methacrylate copolymers, ethoxyethyl meth-acrylates, cyanoethyl methacrylate, amino alkyl methacrylate copolymer, poly(acrylic acid), poly(methacrylic acid), polycyanoacrylates and combinations thereof.

The nanoparticles used with compositions comprising mRNA(s) as described herein can comprise at least one cationic polymer described herein and/or known in the art.

The nanoparticles used with the compositions comprising mRNA(s) as described herein can comprise at least one amine-containing polymer such as, but not limited to polylysine, polyethylene imine, poly(amidoamine) dendrimers and combinations thereof.

The nanoparticles used with the compositions comprising mRNA(s) as described herein can comprise at least one degradable polyester that can contain polycationic side chains. Degradable polyesters include, but are not limited to, poly(serine ester), poly(L-lactide-co-L-lysine), poly(4-hydroxy-L-proline ester), and combinations thereof. In another embodiment, the degradable polyesters can include a PEG conjugation to form a PEGylated polymer.

The compositions comprising mRNA(s) as described herein can be encapsulated in, linked to and/or associated with synthetic nanocarriers. The synthetic nanocarriers can be formulated using methods known in the art and/or described herein. As a non-limiting example, the synthetic nanocarriers can be formulated by the methods described in International Pub Nos. WO2010005740, WO2010030763 and US Pub. Nos. US20110262491, US20100104645 and US20100087337, each of which is herein incorporated by reference in their entireties. In some embodiments, the synthetic nanocarriers can contain reactive groups to release the compositions comprising mRNA(s) as described herein (see International Pub. No. WO20120952552 and US Pub No. US20120171229, each of which is herein incorporated by reference in their entireties).

In some embodiments of any of the aspects, the LNP comprises at least one ionizable lipid, at least one phospholipid, and at least one polyethylene glycol (PEG)-lipid. In some embodiments of any of the aspects, the LNP comprises at least one ionizable lipid, at least one phospholipid, at least one structured lipid, and at least one polyethylene glycol (PEG)-lipid.

Exemplary ionizable lipids include but are not limited to 2,2-dioleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA); dioleyl-methyl 4-dimethylaminobutyrate (DLin-MC3-DMA); and di ((Z)-non-2-en-1-yl) 9-((4-(dimethylamino) butyryl) oxy) heptadecanedioate (L319); ALC-0315; C12-200, cKK-E12; A6; A18-Iso5-2DC18; OF-02; 98N12-5; 9A1P9; C12-200; cKK-E12; 7C1; G0-C 14 ; L319; 3040 13 ; FTT5; 306O 110 ; DODAP; and DODMA. In some embodiments of any of the aspects, the at least one ionizable lipid comprises, consists of, or consists essentially of 2,2-dioleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA); dioleyl-methyl 4-dimethylaminobutyrate (DLin-MC3-DMA); and/or di ((Z)-non-2-en-1-yl) 9-((4-(dimethylamino) butyryl) oxy) heptadecanedioate (L319). In some embodiments of any of the aspects, the at least one ionizable lipid has a pKA in the range of 6.0-6.5.

Exemplary phospholipids include but are not limited to phosphatidic acid (phosphatidate) (PA); phosphatidylethanolamine (cephalin) (PE); phosphatidylcholine (lecithin) (PC); phosphatidylserine (PS): phosphoinositides: phosphatidylinositol (PI); phosphatidylinositol phosphate (PIP); phosphatidylinositol bisphosphate (PIP2); phosphatidylinositol trisphosphate (PIP3); phosphatidic acid derivatives (e.g., DMPA, DPPA, DSPA); phosphatidylcholine derivatives (e.g., DDPC, DLPC, DMPC, DPPC, DSPC, DOPC, POPC, DEPC); phosphatidylglycerol derivatives (e.g., DMPG, DPPG, DSPG, POPG); phosphatidylethanolamine derivatives (e.g., DMPE, DPPE, DSPE DOPE); and phosphatidylserine derivatives (DOPS); and PEG phospholipids (e.g., mPEG-phospholipid, polyglycerin-phospholipid, functionalized-phospholipid, terminal activated-phospholipid). In some embodiments of any of the aspects, the at least one phospholipid comprises, consists of, or consists essentially of phosphatidylcholine.

In some embodiments of any of the aspects, the at least one structured lipid comprises, consists of, or consists essentially of cholesterol.

In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is complexed to a nanoparticle by encapsulation in the interior of the nanoparticle. In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is complexed to a nanoparticle by being interspersed within the lipid bilayer of the nanoparticle. In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is complexed to a nanoparticle by being attached to the nanoparticle via a linking molecule.

›DETAILED DESCRIPTION · 12 of 32

In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is complexed to a LNP by encapsulation in the interior of the LNP. In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is complexed to a LNP by being interspersed within the lipid bilayer of the LNP. In some embodiments of any of the aspects, the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is complexed to a LNP by being attached to the LNP via a linking molecule.

In some embodiments of any of the aspects, the composition further comprises N-acetyl cysteine (NAC). In one aspect of any of the embodiments described herein is a combination of a composition described herein and N-acetyl cysteine (NAC). In one aspect of any of the embodiments described herein is a combination of a first composition comprising a first engineered liver regenerative factor mRNA or liver regenerative factor mRNA described herein and a second composition comprising a second engineered liver regenerative factor mRNA or liver regenerative factor mRNA described herein.

In one aspect, described herein is a kit comprising one or more compositions as described herein, e.g., a composition comprising an engineered liver regenerative factor mRNA and/or liver regenerative factor mRNA, as described herein. A kit is any manufacture (e.g., a package or container) comprising at least one reagent, e.g., an mRNA, the manufacture being promoted, distributed, or sold as a unit for performing the methods described herein.

The kits described herein can optionally comprise additional components useful for performing the methods described herein. By way of example, the kit can comprise fluids (e.g., buffers) suitable for composition comprising an mRNA as described herein, an instructional material which describes performance of a method as described herein, and the like. A kit can further comprise devices and/or reagents for delivery of the composition(s) as described herein. Additionally, the kit may comprise an instruction leaflet and/or may provide information as to the relevance of the obtained results.

The compositions and combinations described herein can promote liver repair and/or healing, or decrease the rate or degree of liver damage and/or injury. Accordingly, in one aspect of any of the embodiments, described herein is a method of treating liver injury or liver disease in a subject in need thereof, the method comprising administering the composition or combination described herein to the subject. In one aspect of any of the embodiments, described herein is a method of accelerating intrinsic liver repair in a subject in need thereof, the method comprising administering the composition or combination described herein.

As used herein, “liver injury or liver disease” refers to injuries or disorders that affect the anatomy or metabolism of normal functioning of the liver. Liver disease can be acute liver disease, chronic liver disease, and/or genetic liver disease. Liver diseases are caused by, for example, alcohol (e.g. ASH), non-alcoholic fatty liver changes (such as NAFLD including NASH), nutrition-mediated liver injury (for example starvation), other toxic liver injury (such as unspecific hepatitis induced by e.g. drugs such as but not limited to acetaminophen (paracetamol), chlorinated hydrocarbons (e.g. CC14), amiodarone (cordarone), valproate, tetracycline (only i.v.), isoniacid, or food intoxication resulting in acute or chronic liver failure, e.g. by consumption of mushrooms containing aflatoxins (preferably B1 aflatoxin) or ingestion of certain metals (such as copper or cadmium) or herbal products used in natural medicine (homeopathics such as Mild thistle, Chaparral, Kawa-Kawa), interference of bilirubin metabolism, hepatitis like syndromes, cholestasis, granulomatous lesions, intrahepatic vascular lesions and cirrhosis), trauma and surgery (e.g. Pringle maneuver), or radiation-mediated liver injury (such as caused by radiotherapy). Exemplary liver injuries and/or liver diseases include but are not limited to haemophilia (including haemophilia A or B), familial hypercholesterolemia, ornithine transcarbamylase deficiency, α-antitrypsin deficiency, non-alcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease, alcohol-related liver disease (ARLD), phenylketonuria, glycogen storage disease, α1-antitrypsin deficiency, hereditary hemochromatosis, tyrosinemia type 1, argininosuccinic aciduria, hepatitis virus infection (e.g., hepatitis A, hepatitis B, or hepatitis C), non-viral hepatitis, autoimmune hepatitis, primary biliary cholangitis, cirrhosis, biliary atresia, liver cancer, genetic cholestasis, hemochromatosis, Gilbert syndrome, primary sclerosing cholangitis (PSC), and Wilson's disease. In some embodiments of any of the aspects, the subject is a subject in need of treatment for acute liver disease, chronic liver disease, or acetaminophen (acetyl-para-aminophenol, APAP) overdose.

The liver is capable of intrinsic repair processes, e.g., restoration of normal hepatic function and architecture. This process is referred to herein as “intrinsic liver repair” and can include inhibition or reversal of fibrosis, decreases in the rate or amount of collagen matrix formation, reduction of inflammatory processes, promotion of health hepatocyte gene expression, and/or promotion of healthy hepatocyte function.

In some embodiments, the methods described herein relate to treating a subject having or diagnosed as having a liver injury or liver disease with a composition or combination comprising mRNA(s) as described herein. Subjects having liver disease can be identified by a physician using current methods of diagnosing liver diseases. Symptoms and/or complications of liver diseases which characterize these conditions and aid in diagnosis are well known in the art and include but are not limited to, jaundice, abdominal pain, bruising, changes in urine or stool color, fatigue, vomiting, or edema. Tests that may aid in a diagnosis of, e.g. liver disease include, but are not limited to, liver enzyme blood tests, ultrasounds, MRI, CT scan, or a liver biopsy. A family history of liver disease, or exposure to risk factors for liver disease (e.g. alcohol consumption) can also aid in determining if a subject is likely to have liver disease or in making a diagnosis of liver disease.

›DETAILED DESCRIPTION · 13 of 32

In some embodiments of any of the aspects, administration or treatment according to the methods described herein can result in decreased serum ALT levels, decreased necrosis, and increased survival following APAP overdose or in response to chronic liver disease, e.g., as compared to absence of said administration or treatment. In some further embodiments of any of the aspects, administration or treatment according to the methods described herein can result in increased conversion of biliary epithelial cells into hepatocytes, decreased steatosis, decreased inflammation, and decreased fibrosis, e.g., as compared to absence of said administration or treatment. In further embodiments of any of the aspects, administration or treatment according to the methods described herein can result in increased hepatocyte proliferation, increased hepatocyte survival, and improvement of hepatocyte transplantation or engraftment, e.g., as compared to absence of said administration or treatment

The compositions and methods described herein can be administered to a subject having or diagnosed as having liver disease. In some embodiments, the methods described herein comprise administering an effective amount of compositions described herein, e.g. an mRNA(s) as described herein to a subject in order to alleviate a symptom of a liver disease. As used herein, “alleviating a symptom” is ameliorating any condition or symptom associated with the disease. As compared with an equivalent untreated control, such reduction is by at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, 99% or more as measured by any standard technique. A variety of means for administering the compositions described herein to subjects are known to those of skill in the art. Such methods can include, but are not limited to oral, parenteral, intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, cutaneous, topical, injection, or intratumoral administration. Administration can be local or systemic.

The compositions and combinations described herein can also promote engraftment of cells in liver tissue. Accordingly, in one aspect of any of the embodiments, described herein is a method of engrafting cells in a liver tissue, the method comprising introducing the cells into the liver tissue and contacting the cells or the liver tissue with a composition or combination described herein.

The introduced cells can be primary cells, differentiated cells, cultured cells, transplanted cells, autologous cells, or non-autologous cells. In some embodiments of any of the aspects, the cells are primary human hepatocytes (PHH). In some embodiments of any of the aspects, the cells are induced pluripotent stem cell-derived hepatocyte-like-cells (iPSC-HLCs).

As used herein, the terms “hepatocytes” and the like refers to cells that show liver-like qualities such as expression of liver genes and proteins and/or hepatic cells. A number of genes expressed in the liver includes carrier protein Albumin (ALB), blood coagulation factors Fibrinogen Alpha chain (FBA), Fibrinogen Beta chain (FBB), Fibrinogen Gamma chain (FBG), thrombinogen, Alpha anti-trypsin (AAT), Tyrosine metabolic genes [Fumarylacetoacetate hydrolase (FAH), tyrosine amino transferase (TAT), homogentisate 1,2-dioxygenase (HGD), 4-hydroxyphenylpyruvic acid dioxygenase (HPD), phenylalanine hydroxylase (PAH), maleylacetoacetate isomerase (MAI)], and urea metabolic genes (ARG1), ornithine carbamoyltransferase (OTC), Carbamoyl-phosphate synthase 1 (CPS1), Glutamine Synthetase (GS). The term “hepatocytes or hepatocyte-like cells” includes different hepatocyte subtypes, which includes but not limited to stem cells, progenitors, differentiated perivenous hepatocytes, and periportal hepatocytes. Furthermore, the term “hepatocyte-like cells” refers to cells that may possess liver or hepatic stem cell or progenitor properties, including but not limited to the capacity to self-renew, proliferate and differentiate. Hepatocyte-like cells may or may not be mature and functional but can engraft into the liver.

In some embodiments of any of the aspects, the liver tissue is a liver in a subject, the introducing comprises transplanting the cells into the liver, and the contacting comprises administering. Accordingly, in one aspect of any of the embodiments, described herein is a method of engrafting cells in a liver of a subject, the method comprising transplanting the cells into the liver and administering to the subject a composition or combination described herein. The administering can be performed before, during, and/or after the transplantation. In some embodiments of any of the aspects, the subject is a subject in need of treatment for acute liver disease, chronic liver disease, or genetic liver disease. In some embodiments of any of the aspects, the subject is a subject in need of treatment for alpha-1 antitrypsin deficiency associated liver disease (AATD).

The term “effective amount” as used herein refers to the amount of a composition or combination described herein needed to alleviate at least one or more symptom of the disease or disorder, and relates to a sufficient amount of pharmacological composition to provide the desired effect. The term “therapeutically effective amount” therefore refers to an amount of a composition or combination that is sufficient to provide a particular effect when administered to atypical subject. An effective amount as used herein, in various contexts, would also include an amount sufficient to delay the development of a symptom of the disease, alter the course of a symptom disease (for example but not limited to, slowing the progression of a symptom of the disease), or reverse a symptom of the disease. Thus, it is not generally practicable to specify an exact “effective amount”. However, for any given case, an appropriate “effective amount” can be determined by one of ordinary skill in the art using only routine experimentation.

Effective amounts, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dosage can vary depending upon the dosage form employed and the route of administration utilized. The dose ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as the ratio LD50/ED50. Compositions and methods that exhibit large therapeutic indices are preferred. A therapeutically effective dose can be estimated initially from cell culture assays. Also, a dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of an mRNA, which achieves a half-maximal inhibition of symptoms) as determined in cell culture, or in an appropriate animal model. Levels in plasma can be measured, for example, by high performance liquid chromatography. The effects of any particular dosage can be monitored by a suitable bioassay, e.g., assay for liver enzymes, among others. The dosage can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment.

›DETAILED DESCRIPTION · 14 of 32

Effective amounts, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the minimal effective dose and/or maximal tolerated dose. The dosage can vary depending upon the dosage form employed and the route of administration utilized. A therapeutically effective dose can be estimated initially from cell culture assays. Also, a dose can be formulated in animal models to achieve a dosage range between the minimal effective dose and the maximal tolerated dose. The effects of any particular dosage can be monitored by a suitable bioassay, e.g., assay for liver function among others. The dosage can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment.

In some embodiments, the technology described herein relates to a pharmaceutical composition comprising mRNA(s) as described herein, and optionally a pharmaceutically acceptable carrier. In some embodiments, the active ingredients of the pharmaceutical composition comprise one or more mRNAs as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist essentially of the one or more mRNAs as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist of one or more mRNAs as described herein. In some embodiments, the active ingredients of the pharmaceutical composition comprise one or more mRNAs as described herein and a carrier (e.g, a LNP). In some embodiments, the active ingredients of the pharmaceutical composition consist essentially of the one or more mRNAs as described herein and a carrier (e.g, a LNP). In some embodiments, the active ingredients of the pharmaceutical composition consist of one or more mRNAs as described herein and a carrier (e.g, a LNP).

Pharmaceutically acceptable carriers and diluents include saline, aqueous buffer solutions, solvents and/or dispersion media. The use of such carriers and diluents is well known in the art. Some non-limiting examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids (23) serum component, such as serum albumin, HDL and LDL; (22) C 2 -C 12 alcohols, such as ethanol; and (23) other non-toxic compatible substances employed in pharmaceutical formulations. Wetting agents, coloring agents, release agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservative and antioxidants can also be present in the formulation. The terms such as “excipient”, “carrier”, “pharmaceutically acceptable carrier” or the like are used interchangeably herein. In some embodiments, the carrier inhibits the degradation of the active agent, e.g. mRNA(s) as described herein.

In some embodiments, the pharmaceutical composition as described herein can be a parenteral dose form. Since administration of parenteral dosage forms typically bypasses the patient's natural defenses against contaminants, parenteral dosage forms are preferably sterile or capable of being sterilized prior to administration to a patient. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions. In addition, controlled-release parenteral dosage forms can be prepared for administration of a patient, including, but not limited to, DUROS®-type dosage forms and dose-dumping.

Suitable vehicles that can be used to provide parenteral dosage forms of mRNA(s) as disclosed within are well known to those skilled in the art. Examples include, without limitation: sterile water; water for injection USP; saline solution; glucose solution; aqueous vehicles such as but not limited to, sodium chloride injection, Ringer's injection, dextrose Injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and propylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate. Compounds that alter or modify the solubility of a pharmaceutically acceptable salt of a compound as disclosed herein can also be incorporated into the parenteral dosage forms of the disclosure, including conventional and controlled-release parenteral dosage forms.

Conventional dosage forms generally provide rapid or immediate drug release from the formulation. Depending on the pharmacology and pharmacokinetics of the drug, use of conventional dosage forms can lead to wide fluctuations in the concentrations of the drug in a patient's blood and other tissues. These fluctuations can impact a number of parameters, such as dose frequency, onset of action, duration of efficacy, maintenance of therapeutic blood levels, toxicity, side effects, and the like. Advantageously, controlled-release formulations can be used to control a drug's onset of action, duration of action, plasma levels within the therapeutic window, and peak blood levels. In particular, controlled- or extended-release dosage forms or formulations can be used to ensure that the maximum effectiveness of a drug is achieved while minimizing potential adverse effects and safety concerns, which can occur both from under-dosing a drug (i.e., going below the minimum therapeutic levels) as well as exceeding the toxicity level for the drug. In some embodiments, the composition or combination can be administered in a sustained release formulation.

›DETAILED DESCRIPTION · 15 of 32

Controlled-release pharmaceutical products have a common goal of improving drug therapy over that achieved by their non-controlled release counterparts. Ideally, the use of an optimally designed controlled-release preparation in medical treatment is characterized by a minimum of drug substance being employed to cure or control the condition in a minimum amount of time. Advantages of controlled-release formulations include: 1) extended activity of the drug; 2) reduced dosage frequency; 3) increased patient compliance; 4) usage of less total drug; 5) reduction in local or systemic side effects; 6) minimization of drug accumulation; 7) reduction in blood level fluctuations; 8) improvement in efficacy of treatment; 9) reduction of potentiation or loss of drug activity; and 10) improvement in speed of control of diseases or conditions. Kim, Cherng-ju, Controlled Release Dosage Form Design, 2 (Technomic Publishing, Lancaster, Pa.: 2000).

Most controlled-release formulations are designed to initially release an amount of drug (active ingredient) that promptly produces the desired therapeutic effect, and gradually and continually release other amounts of drug to maintain this level of therapeutic or prophylactic effect over an extended period of time. In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body. Controlled-release of an active ingredient can be stimulated by various conditions including, but not limited to, pH, ionic strength, osmotic pressure, temperature, enzymes, water, and other physiological conditions or compounds.

A variety of known controlled- or extended-release dosage forms, formulations, and devices can be adapted for use with the salts and compositions of the disclosure. Examples include, but are not limited to, those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,733,566; and 6,365,185 B1; each of which is incorporated herein by reference. These dosage forms can be used to provide slow or controlled-release of one or more active ingredients using, for example, hydroxypropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems (such as OROS® (Alza Corporation, Mountain View, Calif USA)), or a combination thereof to provide the desired release profile in varying proportions.

In some embodiments of any of the aspects, the composition or combination described herein is administered as a monotherapy, e.g., another treatment for the liver disease is not administered to the subject.

In some embodiments of any of the aspects, the methods described herein can further comprise administering a second agent and/or treatment to the subject, e.g. as part of a combinatorial therapy. Non-limiting examples of second agents and/or treatments can include a liver transplant, transjugular intrahepatic portosystemic shunt, anti-virals, or interferon. In some embodiments of any of the aspects, a method described herein can further comprise administering N-acetyl cysteine (NAC) to the subject.

In certain embodiments, an effective dose of a composition or combination as described herein can be administered to a patient once. In certain embodiments, an effective dose of a composition or combination as described herein can be administered to a patient repeatedly. For systemic administration, subjects can be administered a therapeutic amount of a composition or combination as described herein, such as, e.g. 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, or more.

In some embodiments, after an initial treatment regimen, the treatments can be administered on a less frequent basis. For example, after treatment biweekly for three months, treatment can be repeated once per month, for six months or a year or longer. Treatment according to the methods described herein can reduce levels of a marker or symptom of a condition, e.g. liver disease by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80% or at least 90% or more.

The dosage of a composition as described herein can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment. With respect to duration and frequency of treatment, it is typical for skilled clinicians to monitor subjects in order to determine when the treatment is providing therapeutic benefit, and to determine whether to increase or decrease dosage, increase or decrease administration frequency, discontinue treatment, resume treatment, or make other alterations to the treatment regimen. The dosing schedule can vary from once a week to daily depending on a number of clinical factors, such as the subject's sensitivity to the active ingredient. The desired dose or amount of activation can be administered at one time or divided into subdoses, e.g., 2-4 subdoses and administered over a period of time, e.g., at appropriate intervals through the day or other appropriate schedule. In some embodiments, administration can be chronic, e.g., one or more doses and/or treatments daily over a period of weeks or months. Examples of dosing and/or treatment schedules are administration daily, twice daily, three times daily or four or more times daily over a period of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months, or more. A preferred example of dosing and/or treatment schedules are administration every two days, 3 times a week over a period of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months, or more. A composition or combination as described herein can be administered over a period of time, such as over a 5 minute, 10 minute, 15 minute, 20 minute, or 25 minute period.

›DETAILED DESCRIPTION · 16 of 32

The dosage ranges for the administration of a composition or combination described herein, according to the methods described herein depend upon, for example, the form of the mRNA(s), its potency, and the extent to which symptoms, markers, or indicators of a condition described herein are desired to be reduced, for example the percentage increase desired for gene expression or liver function. The dosage should not be so large as to cause adverse side effects. Generally, the dosage will vary with the age, condition, and sex of the patient and can be determined by one of skill in the art. The dosage can also be adjusted by the individual physician in the event of any complication.

The efficacy of a composition or combination described herein in, e.g. the treatment of a condition described herein, or to induce a response as described herein (e.g. liver function and/or repair) can be determined by the skilled clinician. However, a treatment is considered “effective treatment,” as the term is used herein, if one or more of the signs or symptoms of a condition described herein are altered in a beneficial manner, other clinically accepted symptoms are improved, or even ameliorated, or a desired response is induced e.g., by at least 10% following treatment according to the methods described herein. Efficacy can be assessed, for example, by measuring a marker, indicator, symptom, and/or the incidence of a condition treated according to the methods described herein or any other measurable parameter appropriate, e.g. liver function or repair. Efficacy can also be measured by a failure of an individual to worsen as assessed by hospitalization, or need for medical interventions (i.e., progression of the disease is halted). Methods of measuring these indicators are known to those of skill in the art and/or are described herein. Treatment includes any treatment of a disease in an individual or an animal (some non-limiting examples include a human or an animal) and includes: (1) inhibiting the disease, e.g., preventing a worsening of symptoms (e.g. pain or inflammation); or (2) relieving the severity of the disease, e.g., causing regression of symptoms. An effective amount for the treatment of a disease means that amount which, when administered to a subject in need thereof, is sufficient to result in effective treatment as that term is defined herein, for that disease. Efficacy of an agent can be determined by assessing physical indicators of a condition or desired response, (e.g. liver repair and/or function). It is well within the ability of one skilled in the art to monitor efficacy of administration and/or treatment by measuring any one of such parameters, or any combination of parameters. Efficacy can be assessed in animal models of a condition described herein, for example treatment of liver disease. When using an experimental animal model, efficacy of treatment is evidenced when a statistically significant change in a marker is observed, e.g. liver enzymes, liver histology normalization, or hepatocyte gene expression.

In vitro and animal model assays are provided herein which allow the assessment of a given dose of one or more mRNA(s) as described herein. By way of non-limiting example, the effects of a dose of one or more mRNA(s) as described herein can be assessed by in vitro assays for liver cell function, or an animal model, e.g., a murine model as described herein.

In one respect, the present invention relates to the herein described compositions, methods, and respective component(s) thereof, as essential to the technology, yet open to the inclusion of unspecified elements, essential or not (“comprising). In some embodiments of any of the aspects, other elements to be included in the description of the composition, method or respective component thereof are limited to those that do not materially affect the basic and novel characteristic(s) of the technology (e.g., the composition, method, or respective component thereof “consists essentially of” the elements described herein). This applies equally to steps within a described method as well as compositions and components therein. In other embodiments of any of the aspects, the compositions, methods, and respective components thereof, described herein are intended to be exclusive of any element not deemed an essential element to the component, composition or method (e.g., the composition, method, or respective component thereof “consists of” the elements described herein). This applies equally to steps within a described method as well as compositions and components therein.

For convenience, the meaning of some terms and phrases used in the specification, examples, and appended claims, are provided below. Unless stated otherwise, or implicit from context, the following terms and phrases include the meanings provided below. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is an apparent discrepancy between the usage of a term in the art and its definition provided herein, the definition provided within the specification shall prevail.

Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should be understood that this invention is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary. The terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is defined solely by the claims. Definitions of common terms in immunology and molecular biology can be found in The Merck Manual of Diagnosis and Therapy, 19th Edition, published by Merck Sharp & Dohme Corp., 2011 (ISBN 978-0-911910-19-3); Robert S. Porter et al. (eds.), Fields Virology, 6 th Edition, published by Lippincott Williams & Wilkins, Philadelphia, PA, USA (2013), Knipe, D. M. and Howley, P. M. (ed.), The Encyclopedia of Molecular Cell Biology and Molecular Medicine, published by Blackwell Science Ltd., 1999-2012 (ISBN 9783527600908); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8); Immunology by Werner Luttmann, published by Elsevier, 2006; Janeway's Immunobiology, Kenneth Murphy, Allan Mowat, Casey Weaver (eds.), Taylor & Francis Limited, 2014 (ISBN 0815345305, 9780815345305); Lewin's Genes XI, published by Jones & Bartlett Publishers, 2014 (ISBN-1449659055); Michael Richard Green and Joseph Sambrook, Molecular Cloning: A Laboratory Manual, 4 th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA (2012) (ISBN 1936113414); Davis et al., Basic Methods in Molecular Biology, Elsevier Science Publishing, Inc., New York, USA (2012) (ISBN 044460149X); Laboratory Methods in Enzymology: DNA, Jon Lorsch (ed.) Elsevier, 2013 (ISBN 0124199542); Current Protocols in Molecular Biology (CPMB), Frederick M. Ausubel (ed.), John Wiley and Sons, 2014 (ISBN047150338X, 9780471503385), Current Protocols in Protein Science (CPPS), John E. Coligan (ed.), John Wiley and Sons, Inc., 2005; and Current Protocols in Immunology (CPI) (John E. Coligan, ADA M Kruisbeek, David H Margulies, Ethan M Shevach, Warren Strobe, (eds.) John Wiley and Sons, Inc., 2003 (ISBN 0471142735, 9780471142737), the contents of which are all incorporated by reference herein in their entireties.

›DETAILED DESCRIPTION · 17 of 32

The terms “decrease”, “reduce”, “inhibit”, or other grammatical forms thereof are used herein to mean a decrease by a statistically significant amount. In some embodiments, “reduce” or “decrease” or “inhibit” typically means a decrease by at least 10% as compared to a reference level (e.g. the absence of a given treatment or agent) and can include, for example, a decrease by at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more. As used herein, “inhibition” does not encompass a complete inhibition as compared to a reference level. “Complete inhibition” is a 100% inhibition as compared to a reference level. Where applicable, a decrease can be preferably down to a level accepted as within the range of normal for a subject without a given disease (e.g., liver disease).

The terms “increased”, “increase”, “enhance”, or grammatical forms thereof are used herein to mean an increase by a statically significant amount. In some embodiments, the terms “increased”, “increase”, or “enhance”, can mean an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level.

As used herein, a “subject” means a human or animal. Usually the animal is a vertebrate such as a primate, rodent, domestic animal or game animal. Primates include, for example, chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, e.g., Rhesus. Rodents include, for example, mice, rats, woodchucks, ferrets, rabbits and hamsters. Domestic and game animals include, for example, cows, horses, pigs, deer, bison, feline species, e.g., domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon. In some embodiments, the subject is a mammal, e.g., a primate, e.g., a human.

Preferably, the subject is a mammal. The mammal can be a human, non-human primate, mouse, rat, dog, cat, horse, or cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects that represent animal models of disease e.g., cancer. A subject can be male or female.

A subject can be one who has been previously diagnosed with or identified as suffering from or having a condition in need of treatment (e.g. chronic or acute liver injury or disease) or one or more complications related to such a condition, and optionally, have already undergone treatment for the condition or the one or more complications related to the condition. Alternatively, a subject can also be one who has not been previously diagnosed as having such condition or related complications. For example, a subject can be one who exhibits one or more risk factors for the condition or one or more complications related to the condition or a subject who does not exhibit risk factors.

A “subject in need” of treatment for a particular condition can be a subject having that condition, diagnosed as having that condition, or at risk of developing that condition.

As used herein, the terms “protein” and “polypeptide” are used interchangeably herein to designate a series of amino acid residues, connected to each other by peptide bonds between the alpha-amino and carboxyl groups of adjacent residues. The terms “protein”, and “polypeptide” refer to a polymer of amino acids, including modified amino acids (e.g., phosphorylated, glycated, glycosylated, etc.) and amino acid analogs, regardless of its size or function. “Protein” and “polypeptide” are often used in reference to relatively large polypeptides, whereas the term “peptide” is often used in reference to small polypeptides, but usage of these terms in the art overlaps. The terms “protein” and “polypeptide” are used interchangeably herein when referring to a gene product and fragments thereof. Thus, exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments and other equivalents, variants, fragments, and analogs of the foregoing.

As used herein, the term “engineered” and its grammatical equivalents can refer to one or more human-designed and human-generated alterations of a nucleic acid, e.g., a mRNA. The term can refer to alterations, additions, and/or deletion of nucleotides. For example, a mRNA is considered to be “engineered” when at least one aspect of the mRNA, e.g., its sequence, has been manipulated by the hand of man to differ from the aspect as it exists in nature.

The terms “nucleic acid segment,” “nucleotide sequence,” or more generally “segment” will be understood by those in the art as a functional term that includes both genomic sequences, ribosomal RNA sequences, transfer RNA sequences, messenger RNA sequences, small regulatory RNAs, operon sequences and smaller engineered nucleotide sequences that express or may be adapted to express, proteins, polypeptides or peptides. Nucleic acids of the present disclosure may also be synthesized, either completely or in part, by methods known in the art. Thus, all or a portion of the nucleic acids of the present codons may be synthesized using codons preferred by a selected host. Species-preferred codons may be determined, for example, from the codons used most frequently in the proteins expressed in a particular host species. Other modifications of the nucleotide sequences may result in mutants having slightly altered activity

›DETAILED DESCRIPTION · 18 of 32

As used herein, the term “nucleic acid” refers to a single or double-stranded polymer of deoxyribonucleotide or ribonucleotide bases read from the 5′ to the 3′ end. The “nucleic acid” may also optionally contain non-naturally occurring or altered nucleotide bases that permit correct read-through by a polymerase and do not reduce expression of a polypeptide encoded by that nucleic acid. The term “nucleotide sequence” or “nucleic acid sequence” refers to both the sense and antisense strands of a nucleic acid as either individual single strands or in the duplex. The term “ribonucleic acid” (RNA) is inclusive of RNAi (inhibitory RNA), dsRNA (double stranded RNA), siRNA (small interfering RNA), mRNA (messenger RNA), miRNA (micro-RNA), tRNA (transfer RNA, whether charged or discharged with a corresponding acylated amino acid), and cRNA (complementary RNA) and the term “deoxyribonucleic acid” (DNA) is inclusive of cDNA and genomic DNA and DNA-RNA hybrids.

The term “constitutive” when made in reference to a promoter means that the promoter is capable of directing transcription of an operably linked nucleic acid sequence in the absence of a stimulus (e.g., heat shock, chemicals, light, etc.). Typically, constitutive promoters are capable of directing expression of a transgene in substantially any cell and any tissue. In contrast, a “regulatable” or “inducible” promoter is one which is capable of directing a level of transcription of an operably linked nuclei acid sequence in the presence of a stimulus (e.g., heat shock, chemicals, light, etc.) which is different from the level of transcription of the operably linked nucleic acid sequence in the absence of the stimulus.

The term “expression” refers to the cellular processes involved in producing RNA and proteins and as appropriate, secreting proteins, including where applicable, but not limited to, for example, transcription, transcript processing, translation and protein folding, modification and processing. Expression can refer to the transcription and stable accumulation of sense (mRNA) or antisense RNA derived from a nucleic acid fragment or fragments of the invention and/or to the translation of mRNA into a polypeptide.

In some embodiments, the expression of a biomarker(s), target(s), or gene/polypeptide described herein is/are tissue-specific. In some embodiments, the expression of a biomarker(s), target(s), or gene/polypeptide described herein is/are global. In some embodiments, the expression of a biomarker(s), target(s), or gene/polypeptide described herein is systemic.

“Expression products” include RNA transcribed from a gene, and polypeptides obtained by translation of mRNA transcribed from a gene. The term “gene” means the nucleic acid sequence which is transcribed (DNA) to RNA in vitro or in vivo when operably linked to appropriate regulatory sequences. The gene may or may not include regions preceding and following the coding region, e.g. 5′ untranslated (5′UTR) or “leader” sequences and 3′ UTR or “trailer” sequences, as well as intervening sequences (introns) between individual coding segments (exons).

“Operably linked” refers to an arrangement of elements wherein the components so described are configured so as to perform their usual function. Thus, control elements operably linked to a coding sequence are capable of effecting the expression of the coding sequence. The control elements need not be contiguous with the coding sequence, so long as they function to direct the expression thereof. Thus, for example, intervening untranslated yet transcribed sequences can be present between a promoter sequence and the coding sequence and the promoter sequence can still be considered “operably linked” to the coding sequence.

In some embodiments of any of the aspects, the mRNA(s) described herein is exogenous. In some embodiments of any of the aspects, the mRNA(s) described herein is ectopic. In some embodiments of any of the aspects, the mRNA(s) described herein is not endogenous.

The term “exogenous” refers to a substance present in a cell other than its native source. The term “exogenous” when used herein can refer to a nucleic acid (e.g. a nucleic acid encoding a polypeptide) or a polypeptide that has been introduced by a process involving the hand of man into a biological system such as a cell or organism in which it is not normally found and one wishes to introduce the nucleic acid or polypeptide into such a cell or organism. Alternatively, “exogenous” can refer to a nucleic acid or a polypeptide that has been introduced by a process involving the hand of man into a biological system such as a cell or organism in which it is found in relatively low amounts and one wishes to increase the amount of the nucleic acid or polypeptide in the cell or organism, e.g., to create ectopic expression or levels. In contrast, the term “endogenous” refers to a substance that is native to the biological system or cell. As used herein, “ectopic” refers to a substance that is found in an unusual location and/or amount. An ectopic substance can be one that is normally found in a given cell, but at a much lower amount and/or at a different time. Ectopic also includes substance, such as a polypeptide or nucleic acid that is not naturally found or expressed in a given cell in its natural environment.

By “wild-type” it is intended, as will be clear to the skilled addressee, that the nucleic acid or polypeptide is not engineered and can be found in nature. Reference wild-type sequences are provided for the liver regenerative factors described herein.

In some embodiments, the methods described herein relate to measuring, detecting, or determining the level of at least one expression product. As used herein, the term “detecting” or “measuring” refers to observing a signal from, e.g. a probe, label, or target molecule to indicate the presence of an analyte in a sample. Any method known in the art for detecting a particular label moiety can be used for detection. Exemplary detection methods include, but are not limited to, spectroscopic, fluorescent, photochemical, biochemical, immunochemical, electrical, optical or chemical methods. In some embodiments of any of the aspects, measuring can be a quantitative observation.

›DETAILED DESCRIPTION · 19 of 32

As used herein, the terms “treat,” “treatment,” “treating,” or “amelioration” refer to therapeutic treatments, wherein the object is to reverse, alleviate, ameliorate, inhibit, slow down or stop the progression or severity of a condition associated with a disease or disorder. The term “treating” includes reducing or alleviating at least one adverse effect or symptom of a condition, disease or disorder. Treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective” if the progression of a disease is reduced or halted. That is, “treatment” includes not just the improvement of symptoms or markers, but also a cessation of, or at least slowing of, progress or worsening of symptoms compared to what would be expected in the absence of treatment. Beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptom(s), diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, remission (whether partial or total), and/or decreased mortality, whether detectable or undetectable. The term “treatment” of a disease also includes providing relief from the symptoms or side-effects of the disease (including palliative treatment).

In some embodiments of any of the aspects, described herein is a prophylactic method of treatment. As used herein “prophylactic” refers to the timing and intent of a treatment relative to a disease or symptom, that is, the treatment is administered prior to clinical detection or diagnosis of that particular disease or symptom in order to protect the patient from the disease or symptom. Prophylactic treatment can encompass a reduction in the severity or speed of onset of the disease or symptom, or contribute to faster recovery from the disease or symptom. Accordingly, the methods described herein can be prophylactic relative to liver failure, or the need for a liver transplant. In some embodiments of any of the aspects, prophylactic treatment is not prevention of all symptoms or signs of a disease.

As used herein, the term “pharmaceutical composition” refers to the active agent in combination with a pharmaceutically acceptable carrier e.g. a carrier commonly used in the pharmaceutical industry. The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. In some embodiments of any of the aspects, a pharmaceutically acceptable carrier can be a carrier other than water. In some embodiments of any of the aspects, a pharmaceutically acceptable carrier can be a cream, emulsion, gel, liposome, nanoparticle, and/or ointment. In some embodiments of any of the aspects, a pharmaceutically acceptable carrier can be an artificial or engineered carrier, e.g., a carrier that the active ingredient would not be found to occur in in nature.

As used herein, the term “administering,” refers to the placement of a compound as disclosed herein into a subject by a method or route which results in at least partial delivery of the agent at a desired site. Pharmaceutical compositions comprising the compounds disclosed herein can be administered by any appropriate route which results in an effective treatment in the subject. In some embodiments, administration comprises physical human activity, e.g., an injection, act of ingestion, an act of application, and/or manipulation of a delivery device or machine. Such activity can be performed, e.g., by a medical professional and/or the subject being treated.

As used herein, “contacting” refers to any suitable means for delivering, or exposing, an agent to at least one cell. Exemplary delivery methods include, but are not limited to, direct delivery to cell culture medium, perfusion, injection, or other delivery method well known to one skilled in the art. In some embodiments, contacting comprises physical human activity, e.g., an injection; an act of dispensing, mixing, and/or decanting; and/or manipulation of a delivery device or machine.

The term “statistically significant” or “significantly” refers to statistical significance and generally means a two standard deviation (2SD) or greater difference.

Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term “about.” The term “about” when used in connection with percentages can mean±1%.

As used herein, the term “comprising” means that other elements can also be present in addition to the defined elements presented. The use of “comprising” indicates inclusion rather than limitation.

The term “consisting of” refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment.

As used herein the term “consisting essentially of” refers to those elements required for a given embodiment. The term permits the presence of additional elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment of the invention.

As used herein, where a modification (e.g. a change, addition, or deletion) is referred to as “relative to” a reference sequence, it is understood that the modification is made at a point or in a residue of an embodiment of the mRNA that corresponds to the indicated location or residue in the reference sequence. The embodied mRNA can have additional modifications, or may be shorter or longer than the reference sequence, so that the modification does not occur at a location having the same numbering as the location in the reference sequence. One of skill in the art can align sequences and determine what positions or residues correspond to a reference sequence position or residue utilizing automated alignment tools such as BLAST.

›DETAILED DESCRIPTION · 20 of 32

As used herein, the term “corresponding to” refers to an amino acid or nucleotide at the enumerated position in a first polypeptide or nucleic acid, or an amino acid or nucleotide that is equivalent to an enumerated amino acid or nucleotide in a second polypeptide or nucleic acid. Equivalent enumerated amino acids or nucleotides can be determined by alignment of candidate sequences using degree of homology programs known in the art, e.g., BLAST.

The singular terms “a,” “an,” and “the” include plural referents unless context clearly indicates otherwise. Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. The abbreviation, “e.g.” is derived from the Latin exempli gratia, and is used herein to indicate a non-limiting example. Thus, the abbreviation “e.g.” is synonymous with the term “for example.”

In some embodiments of any of the aspects, the disclosure described herein does not concern a process for cloning human beings, processes for modifying the germ line genetic identity of human beings, uses of human embryos for industrial or commercial purposes or processes for modifying the genetic identity of animals which are likely to cause them suffering without any substantial medical benefit to man or animal, and also animals resulting from such processes.

Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is herein deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

Other terms are defined herein within the description of the various aspects of the invention.

All patents and other publications; including literature references, issued patents, published patent applications, and co-pending patent applications; cited throughout this application are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodologies described in such publications that might be used in connection with the technology described herein. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the inventors and does not constitute any admission as to the correctness of the dates or contents of these documents.

The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts of the above references and application to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

Specific elements of any of the foregoing embodiments can be combined or substituted for elements in other embodiments. Furthermore, while advantages associated with certain embodiments of the disclosure have been described in the context of these embodiments, other embodiments may also exhibit such advantages, and not all embodiments need necessarily exhibit such advantages to fall within the scope of the disclosure.

In some embodiments, the present technology may be defined in any of the following numbered paragraphs:

1. A composition comprising at least one engineered liver regenerative factor mRNA, the at least one engineered liver regenerative factor mRNA comprising one or more of:

a) a sequence encoding Growth Hormone (GH) and comprising one or more of the following modifications relative to SEQ ID NO: 1:

deletion of nucleotides 1-63; T→C modification of nucleotide 69; A→C modification of nucleotide 72; G→C modification of nucleotide 81; G→C modification of nucleotide 84; C→G modification of nucleotide 92; T→C modification of nucleotide 99; T→C modification of nucleotide 102; C→G modification of nucleotide 111; T→G modification of nucleotide 126; A→G modification of nucleotide 129; A→T modification of nucleotide 136; G→C modification of nucleotide 137; T→C modification of nucleotide 138; A→C modification of nucleotide 147; T→C modification of nucleotide 153; T→C modification of nucleotide 157; A→G modification of nucleotide 159; A→C modification of nucleotide 163; G→C modification of nucleotide 165; T→G modification of nucleotide 168; T→C modification of nucleotide 171; T→C modification of nucleotide 180; C→G modification of nucleotide 186; T→C modification of nucleotide 195; T→C modification of nucleotide 198; T→C modification of nucleotide 216; T→C modification of nucleotide 234; A→G modification of nucleotide 237; A→G modification of nucleotide 240; T→C modification of nucleotide 246; A→C modification of nucleotide 252; A→G modification of nucleotide 258; T→C modification of nucleotide 267; A→C modification of nucleotide 270; C→G modification of nucleotide 297; T→C modification of nucleotide 300; A→C modification of nucleotide 306; T→C modification of nucleotide 312; T→C modification of nucleotide 315; G→C modification of nucleotide 318; A→C modification of nucleotide 321; A→C modification of nucleotide 331; G→C modification of nucleotide 333; A→G modification of nucleotide 339; A→C modification of nucleotide 342; A→G modification of nucleotide 345; A→G modification of nucleotide 351; A→G modification of nucleotide 360; C→G modification of nucleotide 369; C→G modification of nucleotide 387; G→C modification of nucleotide 396; C→G modification of nucleotide 420; A→C modification of nucleotide 421; G→C modification of nucleotide 423; A→T modification of nucleotide 424; G→C modification of nucleotide 425; T→C modification of nucleotide 426; C→G modification of nucleotide 429; A→T modification of nucleotide 439; G→C modification of nucleotide 440; T→C modification of nucleotide 459; A→T modification of nucleotide 463; G→C modification of nucleotide 464; C→G modification of nucleotide 471; T→C modification of nucleotide 474; C→G modification of nucleotide 480; A→G modification of nucleotide 483; A→G modification of nucleotide 492; A→G modification of nucleotide 498; A→G modification of nucleotide 507; G→C modification of nucleotide 510; G→C modification of nucleotide 519; A→C modification of nucleotide 520; G→C modification of nucleotide 522; A→G modification of nucleotide 528; T→C modification of nucleotide 531; A→T modification of nucleotide 535; G→C modification of nucleotide 536; G→C modification of nucleotide 543; T→C modification of nucleotide 546; G→C modification of nucleotide 549; A→T modification of nucleotide 571; G→C modification of nucleotide 572; A→C modification of nucleotide 585; A→C modification of nucleotide 591; T→C modification of nucleotide 600; A→C modification of nucleotide 606; A→G modification of nucleotide 609; C→G modification of nucleotide 612; G→C modification of nucleotide 624; C→G modification of nucleotide 630; A→C modification of nucleotide 640; G→C modification of nucleotide 642; C→G modification of nucleotide 660; A→C modification of nucleotide 666; T→C modification of nucleotide 693; A→T modification of nucleotide 703; G→C modification of nucleotide 704; T→C modification of nucleotide 708; and deletion of nucleotides 717-823;

›DETAILED DESCRIPTION · 21 of 32

b) a sequence encoding Epidermal Growth Factor (EGF) and comprising one or more of the following modifications relative to SEQ ID NO: 2:

deletion of nucleotides 1-453; C→G modification of nucleotide 462; T→C modification of nucleotide 465; T→G modification of nucleotide 468; T→C modification of nucleotide 474; T→C modification of nucleotide 478; A→C modification of nucleotide 483; A→G modification of nucleotide 486; T→G modification of nucleotide 489; A→C modification of nucleotide 492; A→G modification of nucleotide 495; T→C modification of nucleotide 498; A→T modification of nucleotide 499; G→C modification of nucleotide 500; T→C modification of nucleotide 501; T→C modification of nucleotide 504; T→G modification of nucleotide 507; A→T modification of nucleotide 508; G→C modification of nucleotide 509; T→C modification of nucleotide 510; G→C modification of nucleotide 513; A→C modification of nucleotide 516; A→C modification of nucleotide 519; G→C modification of nucleotide 522; A→T modification of nucleotide 532; G→C modification of nucleotide 533; T→C modification of nucleotide 537; T→C modification of nucleotide 540; A→G modification of nucleotide 543; T→C modification of nucleotide 546; T→C modification of nucleotide 549; C→G modification of nucleotide 552; A→C modification of nucleotide 555; A→C modification of nucleotide 558; T→C modification of nucleotide 561; G→C modification of nucleotide 564; T→C modification of nucleotide 567; T→C modification of nucleotide 570; T→C modification of nucleotide 573; T→C modification of nucleotide 576; T→C modification of nucleotide 582; T→C modification of nucleotide 585; A→C modification of nucleotide 588; T→C modification of nucleotide 595; A→G modification of nucleotide 597; T→C modification of nucleotide 600; T→C modification of nucleotide 609; A→C modification of nucleotide 612; T→C modification of nucleotide 615; A→T modification of nucleotide 616; G→C modification of nucleotide 617; T→C modification of nucleotide 618; T→C modification of nucleotide 624; A→C modification of nucleotide 625; G→C modification of nucleotide 627; T→C modification of nucleotide 630; A→C modification of nucleotide 636; A→G modification of nucleotide 639; A→C modification of nucleotide 642; T→C modification of nucleotide 648; T→C modification of nucleotide 651; A→G modification of nucleotide 657; T→C modification of nucleotide 658; T→C modification of nucleotide 669; T→C modification of nucleotide 672; T→C modification of nucleotide 675; C→G modification of nucleotide 678; A→C modification of nucleotide 681; T→C modification of nucleotide 693; T→C modification of nucleotide 696; T→C modification of nucleotide 699; T→C modification of nucleotide 702; T→C modification of nucleotide 705; A→G modification of nucleotide 711; A→C modification of nucleotide 712; A→C modification of nucleotide 714; T→C modification of nucleotide 720; T→C modification of nucleotide 729; T→C modification of nucleotide 730; A→G modification of nucleotide 732; A→G modification of nucleotide 735; A→C modification of nucleotide 736; A→C modification of nucleotide 738; A→G modification of nucleotide 741; T→G modification of nucleotide 744; T→C modification of nucleotide 745; A→G modification of nucleotide 750; A→C modification of nucleotide 751; A→C modification of nucleotide 753; T→G modification of nucleotide 756; T→C modification of nucleotide 759; T→C modification of nucleotide 765; G→C modification of nucleotide 768; A→C modification of nucleotide 771; A→C modification of nucleotide 772; G→C modification of nucleotide 774; A→G modification of nucleotide 777; A→C modification of nucleotide 781; A→C modification of nucleotide 783; A→G modification of nucleotide 786; T→C modification of nucleotide 789; T→C modification of nucleotide 792; A→C modification of nucleotide 795; A→G modification of nucleotide 801; T→C modification of nucleotide 804; T→G modification of nucleotide 807; T→C modification of nucleotide 810; A→C modification of nucleotide 813; A→C modification of nucleotide 819; A→C modification of nucleotide 822; T→C modification of nucleotide 825; A→C modification of nucleotide 831; T→C modification of nucleotide 834; A→G modification of nucleotide 837; A→G modification of nucleotide 840; T→G modification of nucleotide 843; T→C modification of nucleotide 846; A→C modification of nucleotide 852; T→C modification of nucleotide 855; A→G modification of nucleotide 858; A→G modification of nucleotide 864; A→C modification of nucleotide 867; T→C modification of nucleotide 873; A→C modification of nucleotide 876; A→G modification of nucleotide 879; A→C modification of nucleotide 882; T→C modification of nucleotide 885; A→G modification of nucleotide 891; A→C modification of nucleotide 894; T→C modification of nucleotide 897; T→C modification of nucleotide 900; T→C modification of nucleotide 909; T→G modification of nucleotide 912; T→C modification of nucleotide 913; A→G modification of nucleotide 915; A→T modification of nucleotide 916; G→C modification of nucleotide 917; T→C modification of nucleotide 918; T→C modification of nucleotide 921; T→C modification of nucleotide 922; A→G modification of nucleotide 924; A→G modification of nucleotide 927; T→C modification of nucleotide 930; T→C modification of nucleotide 933; A→C modification of nucleotide 936; T→C modification of nucleotide 939; A→G modification of nucleotide 942; A→C modification of nucleotide 945; T→G modification of nucleotide 948; T→C modification of nucleotide 951; A→C modification of nucleotide 954; A→G modification of nucleotide 957; A→G modification of nucleotide 960; A→C modification of nucleotide 961; T→C modification of nucleotide 966; A→C modification of nucleotide 972; T→C modification of nucleotide 978; A→C modification of nucleotide 981; T→C modification of nucleotide 990; A→C modification of nucleotide 993; A→T modification of nucleotide 994; G→C modification of nucleotide 995; T→G modification of nucleotide 999; T→C modification of nucleotide 1002; A→C modification of nucleotide 1003; A→C modification of nucleotide 1005; A→C modification of nucleotide 1008; T→C modification of nucleotide 1011; C→G modification of nucleotide 1014; T→C modification of nucleotide 1017; T→C modification of nucleotide 1020; A→C modification of nucleotide 1026; T→C modification of nucleotide 1035; T→C modification of nucleotide 1039; A→C modification of nucleotide 1047; A→C modification of nucleotide 1050; A→G modification of nucleotide 1056; A→C modification of nucleotide 1059; A→C modification of nucleotide 1062; T→C modification of nucleotide 1065; A→C modification of nucleotide 1071; T→C modification of nucleotide 1072; T→C modification of nucleotide 1077; T→G modification of nucleotide 1083; T→C modification of nucleotide 1086; G→C modification of nucleotide 1092; T→C modification of nucleotide 1098; T→C modification of nucleotide 1104; A→C modification of nucleotide 1114; A→C modification of nucleotide 1116; A→G modification of nucleotide 1119; A→C modification of nucleotide 1122; A→T modification of nucleotide 1123; G→C modification of nucleotide 1124; T→C modification of nucleotide 1128; T→C modification of nucleotide 1131; T→G modification of nucleotide 1134; T→C modification of nucleotide 1137; T→C modification of nucleotide 1146; T→C modification of nucleotide 1149; T→C modification of nucleotide 1152; T→C modification of nucleotide 1155; A→C modification of nucleotide 1158; T→C modification of nucleotide 1161; T→C modification of nucleotide 1164; C→G modification of nucleotide 1167; T→C modification of nucleotide 1173; A→T modification of nucleotide 1174; G→C modification of nucleotide 1175; T→C modification of nucleotide 1176; A→G modification of nucleotide 1179; T→C modification of nucleotide 1182; A→C modification of nucleotide 1185; A→C modification of nucleotide 1188; T→C modification of nucleotide 1194; T→C modification of nucleotide 1197; T→C modification of nucleotide 1198; T→C modification of nucleotide 1203; A→C modification of nucleotide 1206; T→G modification of nucleotide 1215; T→C modification of nucleotide 1218; T→C modification of nucleotide 1221; T→C modification of nucleotide 1227; T→C modification of nucleotide 1236; A→C modification of nucleotide 1239; A→C modification of nucleotide 1242; A→G modification of nucleotide 1248; A→C modification of nucleotide 1257; T→C modification of nucleotide 1260; A→C modification of nucleotide 1266; A→G modification of nucleotide 1275; T→C modification of nucleotide 1281; A→C modification of nucleotide 1284; T→G modification of nucleotide 1296; A→C modification of nucleotide 1297; A→C modification of nucleotide 1299; T→C modification of nucleotide 1302; C→G modification of nucleotide 1308; T→C modification of nucleotide 1311; A→C modification of nucleotide 1314; 1→C modification of nucleotide 1317; T→C modification of nucleotide 1320; A→G modification of nucleotide 1323; A→C modification of nucleotide 1326; T→G modification of nucleotide 1329; T→C modification of nucleotide 1332; A→G modification of nucleotide 1335; A→G modification of nucleotide 1341; A→G modification of nucleotide 1344; T→C modification of nucleotide 1350; A→C modification of nucleotide 1353; T→G modification of nucleotide 1356; A→C modification of nucleotide 1359; A→G modification of nucleotide 1362; A→C modification of nucleotide 1371; A→G modification of nucleotide 1374; T→C modification of nucleotide 1377; T→C modification of nucleotide 1383; T→C modification of nucleotide 1392; A→G modification of nucleotide 1401; T→G modification of nucleotide 1404; A→G modification of nucleotide 1410; A→C modification of nucleotide 1411; A→C modification of nucleotide 1414; G→C modification of nucleotide 1416; A→G modification of nucleotide 1419; A→C modification of nucleotide 1422; A→T modification of nucleotide 1429; G→C modification of nucleotide 1430; A→T modification of nucleotide 1432; G→C modification of nucleotide 1433; T→C modification of nucleotide 1437; T→C modification of nucleotide 1443; G→C modification of nucleotide 1446; A→G modification of nucleotide 1449; C→G modification of nucleotide 1455; A→C modification of nucleotide 1461; T→C modification of nucleotide 1465; T→C modification of nucleotide 1476; A→C modification of nucleotide 1479; A→C modification of nucleotide 1485; A→G modification of nucleotide 1494; A→T modification of nucleotide 1495; G→C modification of nucleotide 1496; T→C modification of nucleotide 1497; A→C modification of nucleotide 1500; G→C modification of nucleotide 1506; T→C modification of nucleotide 1515; A→G modification of nucleotide 1518; T→C modification of nucleotide 1521; T→G modification of nucleotide 1524; T→C modification of nucleotide 1527; A→G modification of nucleotide 1530; T→C modification of nucleotide 1536; T→C modification of nucleotide 1539; T→C modification of nucleotide 1545; T→C modification of nucleotide 1548; T→C modification of nucleotide 1554; T→C modification of nucleotide 1557; T→G modification of nucleotide 1560; G→C modification of nucleotide 1563; T→C modification of nucleotide 1566; A→G modification of nucleotide 1569; T→C modification of nucleotide 1578; A→C modification of nucleotide 1581; T→C modification of nucleotide 1587; G→C modification of nucleotide 1596; T→C modification of nucleotide 1602; A→G modification of nucleotide 1605; A→C modification of nucleotide 1608; T→C modification of nucleotide 1611; T→G modification of nucleotide 1614; T→G modification of nucleotide 1620; T→C modification of nucleotide 1623; T→C modification of nucleotide 1626; G→C modification of nucleotide 1629; A→G modification of nucleotide 1632; A→G modification of nucleotide 1635; T→C modification of nucleotide 1638; T→C modification of nucleotide 1641; A→G modification of nucleotide 1644; T→G modification of nucleotide 1647; T→G modification of nucleotide 1650; T→C modification of nucleotide 1656; A→C modification of nucleotide 1659; T→C modification of nucleotide 1665; T→C modification of nucleotide 1671; A→G modification of nucleotide 1674; A→T modification of nucleotide 1678; G→C modification of nucleotide 1679; T→C modification of nucleotide 1683; T→C modification of nucleotide 1689; T→G modification of nucleotide 1692; A→C modification of nucleotide 1698; A→C modification of nucleotide 1701; A→G modification of nucleotide 1704; T→C modification of nucleotide 1707; T→C modification of nucleotide 1711; A→G modification of nucleotide 1713; T→C modification of nucleotide 1716; T→C modification of nucleotide 1722; T→C modification of nucleotide 1725; A→G modification of nucleotide 1728; A→C modification of nucleotide 1734; T→G modification of nucleotide 1740; A→C modification of nucleotide 1744; A→C modification of nucleotide 1746; T→C modification of nucleotide 1749; G→C modification of nucleotide 1752; A→G modification of nucleotide 1755; A→C modification of nucleotide 1758; T→C modification of nucleotide 1761; A→T modification of nucleotide 1762; G→C modification of nucleotide 1763; T→C modification of nucleotide 1767; T→C modification of nucleotide 1770; A→C modification of nucleotide 1776; T→C modification of nucleotide 1782; T→C modification of nucleotide 1785; T→C modification of nucleotide 1788; A→C modification of nucleotide 1791; T→C modification of nucleotide 1794; A→T modification of nucleotide 1795; G→C modification of nucleotide 1796; C→G modification of nucleotide 1803; T→G modification of nucleotide 1809; T→C modification of nucleotide 1812; T→G modification of nucleotide 1815; A→T modification of nucleotide 1816; G→C modification of nucleotide 1817; A→C modification of nucleotide 1821; A→G modification of nucleotide 1824; A→G modification of nucleotide 1833; T→C modification of nucleotide 1836; T→C modification of nucleotide 1839; T→C modification of nucleotide 1845; T→C modification of nucleotide 1848; G→C modification of nucleotide 1851; T→C modification of nucleotide 1854; A→G modification of nucleotide 1860; A→G modification of nucleotide 1863; T→C modification of nucleotide 1869; A→G modification of nucleotide 1872; A→G modification of nucleotide 1875; G→C modification of nucleotide 1877; T→C modification of nucleotide 1881; A→C modification of nucleotide 1884; T→C modification of nucleotide 1887; A→C modification of nucleotide 1890; A→C modification of nucleotide 1893; A→C modification of nucleotide 1896; A→G modification of nucleotide 1899; A→C modification of nucleotide 1902; T→C modification of nucleotide 1905; T→C modification of nucleotide 1906; T→C modification of nucleotide 1914; T→C modification of nucleotide 1920; T→C modification of nucleotide 1923; A→G modification of nucleotide 1926; T→C modification of nucleotide 1929; T→C modification of nucleotide 1932; A→C modification of nucleotide 1935; T→C modification of nucleotide 1944; T→C modification of nucleotide 1947; T→C modification of nucleotide 1950; A→C modification of nucleotide 1953; A→C modification of nucleotide 1956; T→C modification of nucleotide 1962; A→C modification of nucleotide 1965; T→C modification of nucleotide 1968; C→G modification of nucleotide 1974; A→T modification of nucleotide 1975; G→C modification of nucleotide 1976; A→C modification of nucleotide 1989; T→G modification of nucleotide 1995; T→C modification of nucleotide 1998; A→G modification of nucleotide 2004; T→C modification of nucleotide 2007; T→C modification of nucleotide 2010; T→C modification of nucleotide 2016; A→G modification of nucleotide 2022; T→C modification of nucleotide 2025; A→C modification of nucleotide 2031; T→C modification of nucleotide 2037; T→C modification of nucleotide 2043; A→C modification of nucleotide 2046; A→C modification of nucleotide 2061; A→C modification of nucleotide 2065; A→C modification of nucleotide 2067; T→C modification of nucleotide 2070; T→C modification of nucleotide 2073; T→C modification of nucleotide 2079; T→C modification of nucleotide 2082; A→C modification of nucleotide 2091; A→G modification of nucleotide 2094; A→C modification of nucleotide 2095; G→C modification of nucleotide 2097; T→G modification of nucleotide 2100; T→C modification of nucleotide 2103; A→G modification of nucleotide 2109; A→C modification of nucleotide 2112; A→G modification of nucleotide 2115; T→C modification of nucleotide 2118; A→C modification of nucleotide 2124; A→G modification of nucleotide 2127; T→C modification of nucleotide 2130; T→G modification of nucleotide 2133; T→C modification of nucleotide 2136; T→C modification of nucleotide 2148; T→C modification of nucleotide 2154; A→C modification of nucleotide 2155; A→C modification of nucleotide 2157; T→C modification of nucleotide 2163; A→C modification of nucleotide 2169; A→C modification of nucleotide 2173; A→C modification of nucleotide 2175; G→C modification of nucleotide 2178; A→G modification of nucleotide 2181; T→C modification of nucleotide 2184; T→C modification of nucleotide 2190; A→C modification of nucleotide 2193; A→C modification of nucleotide 2194; G→C modification of nucleotide 2196; A→T modification of nucleotide 2197; G→C modification of nucleotide 2198; T→C modification of nucleotide 2199; T→C modification of nucleotide 2202; T→C modification of nucleotide 2203; A→G modification of nucleotide 2205; T→C modification of nucleotide 2208; G→C modification of nucleotide 2211; A→G modification of nucleotide 2214; T→G modification of nucleotide 2217; A→G modification of nucleotide 2223; A→C modification of nucleotide 2226; T→C modification of nucleotide 2232; T→C modification of nucleotide 2247; A→G modification of nucleotide 2250; A→C modification of nucleotide 2253; A→C modification of nucleotide 2256; A→C modification of nucleotide 2259; T→C modification of nucleotide 2262; T→C modification of nucleotide 2265; T→G modification of nucleotide 2268; T→C modification of nucleotide 2271; A→C modification of nucleotide 2274; A→C modification of nucleotide 2284; A→C modification of nucleotide 2286; T→C modification of nucleotide 2287; A→G modification of nucleotide 2289; T→C modification of nucleotide 2298; T→C modification of nucleotide 2301; A→C modification of nucleotide 2304; G→C modification of nucleotide 2307; T→C modification of nucleotide 2310; T→C modification of nucleotide 2313; A→C modification of nucleotide 2316; A→C modification of nucleotide 2319; T→C modification of nucleotide 2322; A→G modification of nucleotide 2325; A→T modification of nucleotide 2326; G→C modification of nucleotide 2327; T→C modification of nucleotide 2328; T→C modification of nucleotide 2331; C→G modification of nucleotide 2337; A→G modification of nucleotide 2340; T→G modification of nucleotide 2346; T→C modification of nucleotide 2352; T→G modification of nucleotide 2358; A→C modification of nucleotide 2361; T→A modification of nucleotide 2365; G→C modification of nucleotide 2366; T→C modification of nucleotide 2370; T→C modification of nucleotide 2373; A→G modification of nucleotide 2376; A→T modification of nucleotide 2386; G→C modification of nucleotide 2387; T→C modification of nucleotide 2388; A→C modification of nucleotide 2391; A→C modification of nucleotide 2394; G→C modification of nucleotide 2397; T→C modification of nucleotide 2400; T→C modification of nucleotide 2407; A→G modification of nucleotide 2409; T→C modification of nucleotide 2412; T→C modification of nucleotide 2419; T→C modification of nucleotide 2433; T→C modification of nucleotide 2445; T→C modification of nucleotide 2451; A→G modification of nucleotide 2454; T→C modification of nucleotide 2463; T→C modification of nucleotide 2469; T→C modification of nucleotide 2472; A→C modification of nucleotide 2475; A→G modification of nucleotide 2478; A→C modification of nucleotide 2484; A→C modification of nucleotide 2485; A→C modification of nucleotide 2487; T→G modification of nucleotide 2490; T→C modification of nucleotide 2499; T→C modification of nucleotide 2502; A→G modification of nucleotide 2505; T→C modification of nucleotide 2508; A→C modification of nucleotide 2514; T→C modification of nucleotide 2517; T→C modification of nucleotide 2520; A→G modification of nucleotide 2523; A→C modification of nucleotide 2526; T→C modification of nucleotide 2532; T→C modification of nucleotide 2538; T→C modification of nucleotide 2541; A→C modification of nucleotide 2553; T→C modification of nucleotide 2556; T→C modification of nucleotide 2562; A→C modification of nucleotide 2568; A→C modification of nucleotide 2571; A→G modification of nucleotide 2574; G→C modification of nucleotide 2577; A→C modification of nucleotide 2578; A→C modification of nucleotide 2580; A→G modification of nucleotide 2583; A→C modification of nucleotide 2590; G→C modification of nucleotide 2592; T→C modification of nucleotide 2595; A→G modification of nucleotide 2601; T→C modification of nucleotide 2604; A→C modification of nucleotide 2605; A→C modification of nucleotide 2607; A→G modification of nucleotide 2610; T→C modification of nucleotide 2613; A→C modification of nucleotide 2616; A→G modification of nucleotide 2619; A→T modification of nucleotide 2623; G→C modification of nucleotide 2624; A→C modification of nucleotide 2640; A→C modification of nucleotide 2643; T→G modification of nucleotide 2649; T→G modification of nucleotide 2655; T→C modification of nucleotide 2658; A→C modification of nucleotide 2661; T→C modification of nucleotide 2662; A→C modification of nucleotide 2667; A→G modification of nucleotide 2670; A→C modification of nucleotide 2673; A→C modification of nucleotide 2676; A→C modification of nucleotide 2679; T→C modification of nucleotide 2682; T→C modification of nucleotide 2689; A→G modification of nucleotide 2691; T→C modification of nucleotide 2694; A→G modification of nucleotide 2697; A→C modification of nucleotide 2703; T→C modification of nucleotide 2709; A→G modification of nucleotide 2712; T→C modification of nucleotide 2715; T→C modification of nucleotide 2718; A→G modification of nucleotide 2724; A→C modification of nucleotide 2728; G→C modification of nucleotide 2730; T→G modification of nucleotide 2733; A→C modification of nucleotide 2736; T→C modification of nucleotide 2739; T→C modification of nucleotide 2742; T→C modification of nucleotide 2748; G→C modification of nucleotide 2751; T→C modification of nucleotide 2754; T→C modification of nucleotide 2757; A→G modification of nucleotide 2760; T→C modification of nucleotide 2763; T→C modification of nucleotide 2766; A→G modification of nucleotide 2772; A→C modification of nucleotide 2778; T→C modification of nucleotide 2781; G→C modification of nucleotide 2784; A→G modification of nucleotide 2787; G→C modification of nucleotide 2790; T→C modification of nucleotide 2793; T→C modification of nucleotide 2799; T→C modification of nucleotide 2805; T→C modification of nucleotide 2808; T→C modification of nucleotide 2811; T→C modification of nucleotide 2818; A→C modification of nucleotide 2823; T→C modification of nucleotide 2826; T→C modification of nucleotide 2829; A→G modification of nucleotide 2832; T→G modification of nucleotide 2832; T→G modification of nucleotide 2835; T→C modification of nucleotide 2838; A→G modification of nucleotide 2841; A→G modification of nucleotide 2850; A→G modification of nucleotide 2853; A→C modification of nucleotide 2856; A→C modification of nucleotide 2859; T→C modification of nucleotide 2860; T→C modification of nucleotide 2869; T→C modification of nucleotide 2880; A→C modification of nucleotide 2881; A→C modification of nucleotide 2883; A→C modification of nucleotide 2889; A→G modification of nucleotide 2892; T→C modification of nucleotide 2895; T→C modification of nucleotide 2901; A→C modification of nucleotide 2904; A→G modification of nucleotide 2907; T→C modification of nucleotide 2910; A→G modification of nucleotide 2913; A→G modification of nucleotide 2922; T→C modification of nucleotide 2928; A→G modification of nucleotide 2931; A→C modification of nucleotide 2943; T→C modification of nucleotide 2946; A→G modification of nucleotide 2949; T→C modification of nucleotide 2952; T→C modification of nucleotide 2958; T→C modification of nucleotide 2961; T→C modification of nucleotide 2964; A→C modification of nucleotide 2970; A→T modification of nucleotide 2974; G→C modification of nucleotide 2975; T→C modification of nucleotide 2982; T→C modification of nucleotide 2985; G→C modification of nucleotide 2988; T→C modification of nucleotide 2991; T→C modification of nucleotide 2994; A→C modification of nucleotide 2997; A→C modification of nucleotide 3003; T→C modification of nucleotide 3009; A→C modification of nucleotide 3015; T→C modification of nucleotide 3018; T→C modification of nucleotide 3024; T→C modification of nucleotide 3025; A→G modification of nucleotide 3030; A→C modification of nucleotide 3033; T→C modification of nucleotide 3036; T→C modification of nucleotide 3039; G→C modification of nucleotide 3042; T→C modification of nucleotide 3045; A→C modification of nucleotide 3048; A→G modification of nucleotide 3051; A→G modification of nucleotide 3054; T→C modification of nucleotide 3057; T→C modification of nucleotide 3060; T→C modification of nucleotide 3063; A→C modification of nucleotide 3066; T→C modification of nucleotide 3069; A→G modification of nucleotide 3072; T→C modification of nucleotide 3075; T→C modification of nucleotide 3084; C→G modification of nucleotide 3087; A→C modification of nucleotide 3090; T→C modification of nucleotide 3102; A→G modification of nucleotide 3129; T→C modification of nucleotide 3132; T→C modification of nucleotide 3135; T→C modification of nucleotide 3138; C→G modification of nucleotide 3141; G→C modification of nucleotide 3147; A→C modification of nucleotide 3153; A→G modification of nucleotide 3156; A→G modification of nucleotide 3165; A→C modification of nucleotide 3168; T→C modification of nucleotide 3171; G→C modification of nucleotide 3174; T→C modification of nucleotide 3177; T→C modification of nucleotide 3183; T→G modification of nucleotide 3186; T→C modification of nucleotide 3189; T→C modification of nucleotide 3192; T→C modification of nucleotide 3195; A→G modification of nucleotide 3204; G→C modification of nucleotide 3210; A→T modification of nucleotide 3212; A→T modification of nucleotide 3217; G→C modification of nucleotide 3218; T→C modification of nucleotide 3222; A→C modification of nucleotide 3225; T→C modification of nucleotide 3231; A→T modification of nucleotide 3235; G→C modification of nucleotide 3236; A→C modification of nucleotide 3243; T→C modification of nucleotide 3246; A→C modification of nucleotide 3249; A→C modification of nucleotide 3255; T→C modification of nucleotide 3261; T→C modification of nucleotide 3273; T→C modification of nucleotide 3276; A→C modification of nucleotide 3279; T→C modification of nucleotide 3288; A→G modification of nucleotide 3291; A→C modification of nucleotide 3294; A→C modification of nucleotide 3297; T→C modification of nucleotide 3303; T→C modification of nucleotide 3309; T→C modification of nucleotide 3315; T→C modification of nucleotide 3318; A→C modification of nucleotide 3321; T→C modification of nucleotide 3327; C→G modification of nucleotide 3333; A→C modification of nucleotide 3334; G→C modification of nucleotide 3336; A→G modification of nucleotide 3339; T→C modification of nucleotide 3342; T→C modification of nucleotide 3354; A→G modification of nucleotide 3360; A→C modification of nucleotide 3361; A→C modification of nucleotide 3363; T→C modification of nucleotide 3366; T→A modification of nucleotide 3367; G→C modification of nucleotide 3368; T→C modification of nucleotide 3369; T→C modification of nucleotide 3375; A→G modification of nucleotide 3378; T→C modification of nucleotide 3381; T→C modification of nucleotide 3396; G→C modification of nucleotide 3399; C→G modification of nucleotide 3408; T→C modification of nucleotide 3411; T→C modification of nucleotide 3414; T→C modification of nucleotide 3417; T→C modification of nucleotide 3429; T→C modification of nucleotide 3432; A→G modification of nucleotide 3435; A→C modification of nucleotide 3438; T→C modification of nucleotide 3439; T→C modification of nucleotide 3450; A→C modification of nucleotide 3453; T→C modification of nucleotide 3462; T→G modification of nucleotide 3465; T→G modification of nucleotide 3468; G→C modification of nucleotide 3480; A→G modification of nucleotide 3486; T→C modification of nucleotide 3489; A→C modification of nucleotide 3498; A→G modification of nucleotide 3516; T→C modification of nucleotide 3528; G→C modification of nucleotide 3537; C→G modification of nucleotide 3552; T→C modification of nucleotide 3564; C→G modification of nucleotide 3567; T→G modification of nucleotide 3582; C→G modification of nucleotide 3585; C→G modification of nucleotide 3584; C→G modification of nucleotide 3594; C→G modification of nucleotide 3597; A→T modification of nucleotide 3612; G→C modification of nucleotide 3602; G→C modification of nucleotide 3612; A→C modification of nucleotide 3625; G→C modification of nucleotide 3627; T→C modification of nucleotide 3630; A→G modification of nucleotide 3642; G→C modification of nucleotide 3645; A→G modification of nucleotide 3648; A→C modification of nucleotide 3654; T→C modification of nucleotide 3660; T→C modification of nucleotide 3663; T→C modification of nucleotide 3668; G→C modification of nucleotide 3675; A→T modification of nucleotide 3676; G→C modification of nucleotide 3677; A→C modification of nucleotide 3679; A→C modification of nucleotide 3681; T→C modification of nucleotide 3684; A→C modification of nucleotide 3688; G→C modification of nucleotide 3690; A→T modification of nucleotide 3691; G→C modification of nucleotide 3692; T→C modification of nucleotide 3693; A→C modification of nucleotide 3697; G→C modification of nucleotide 3699; T→C modification of nucleotide 3702; T→C modification of nucleotide 3705; T→C modification of nucleotide 3711; T→C modification of nucleotide 3717; G→C modification of nucleotide 3720; T→C modification of nucleotide 3729; T→C modification of nucleotide 3735; A→G modification of nucleotide 3738; T→C modification of nucleotide 3741; T→C modification of nucleotide 3747; T→G modification of nucleotide 3752; A→C modification of nucleotide 3756; A→G modification of nucleotide 3759; A→G modification of nucleotide 3762; A→G modification of nucleotide 3769; C→G modification of nucleotide 3774; T→C modification of nucleotide 3780; G→C modification of nucleotide 3783; T→C modification of nucleotide 3786; A→G modification of nucleotide 3789; A→C modification of nucleotide 3792; T→C modification of nucleotide 3798; T→C modification of nucleotide 3801; T→C modification of nucleotide 3807; A→C modification of nucleotide 3816; A→C modification of nucleotide 3819; T→C modification of nucleotide 3822; G→C modification of nucleotide 3825; A→C modification of nucleotide 3828; A→G modification of nucleotide 3834; A→C modification of nucleotide 3837; T→C modification of nucleotide 3840; A→C modification of nucleotide 3843; A→C modification of nucleotide 3847; G→C modification of nucleotide 3849; T→C modification of nucleotide 3862; A→G modification of nucleotide 3864; T→C modification of nucleotide 3867; A→C modification of nucleotide 3870; A→C modification of nucleotide 3879; A→G modification of nucleotide 3885; T→C modification of nucleotide 3897; A→C modification of nucleotide 3900; A→G modification of nucleotide 3903; A→T modification of nucleotide 3907; G→C modification of nucleotide 3908; T→C modification of nucleotide 3909; T→C modification of nucleotide 3912; T→C modification of nucleotide 3924; A→G modification of nucleotide 3933; A→G modification of nucleotide 3942; A→T modification of nucleotide 3943; G→C modification of nucleotide 3944; T→C modification of nucleotide 3948; T→C modification of nucleotide 3951; T→C modification of nucleotide 3963; G→C modification of nucleotide 3966; A→C modification of nucleotide 3969; T→G modification of nucleotide 3978; A→G modification of nucleotide 3981; G→C modification of nucleotide 3984; T→C modification of nucleotide 3987; C→G modification of nucleotide 3990; T→C modification of nucleotide 4002; T→C modification of nucleotide 4005; C→G modification of nucleotide 4008; A→G modification of nucleotide 4011; A→C modification of nucleotide 4014; T→C modification of nucleotide 4017; A→C modification of nucleotide 4023; T→C modification of nucleotide 4024; A→G modification of nucleotide 4026; A→G modification of nucleotide 4032; A→G modification of nucleotide 4035; A→C modification of nucleotide 4036; G→C modification of nucleotide 4038; A→C modification of nucleotide 4050; A→C modification of nucleotide 4053; A→G modification of nucleotide 4059; T→C modification of nucleotide 4071; and deletion of nucleotides 4076-6388;

›DETAILED DESCRIPTION · 22 of 32

c) a sequence encoding Hepatocyte Growth Factor (HGF) and comprising one or more of the following modifications relative to SEQ ID NO: 3:

deletion of nucleotides 1-76; A→G modification of nucleotide 91; C→G modification of nucleotide 94; T→C modification of nucleotide 118; C→G modification of nucleotide 121; C→G modification of nucleotide 124; T→C modification of nucleotide 130; C→G modification of nucleotide 133; C→G modification of nucleotide 136; C→G modification of nucleotide 142; T→C modification of nucleotide 160; A→C modification of nucleotide 163; A→C modification of nucleotide 169; A→G modification of nucleotide 172; A→C modification of nucleotide 173; G→C modification of nucleotide 175; A→G modification of nucleotide 178; A→C modification of nucleotide 179; A→C modification of nucleotide 181; A→C modification of nucleotide 182; A→C modification of nucleotide 184; T→C modification of nucleotide 187; A→C modification of nucleotide 190; T→C modification of nucleotide 193; T→C modification of nucleotide 196; A→G modification of nucleotide 199; A→G modification of nucleotide 205; A→G modification of nucleotide 208; A→C modification of nucleotide 211; A→C modification of nucleotide 214; T→C modification of nucleotide 220; A→G modification of nucleotide 226; A→G modification of nucleotide 232; A→C modification of nucleotide 235; T→C modification of nucleotide 238; A→C modification of nucleotide 241; A→C modification of nucleotide 244; A→C modification of nucleotide 253; A→G modification of nucleotide 256; A→G modification of nucleotide 262; A→G modification of nucleotide 265; T→C modification of nucleotide 271; T→C modification of nucleotide 274; A→C modification of nucleotide 277; A→G modification of nucleotide 283; T→C modification of nucleotide 286; T→C modification of nucleotide 289; T→C modification of nucleotide 292; A→C modification of nucleotide 293; A→C modification of nucleotide 295; T→C modification of nucleotide 298; T→C modification of nucleotide 301; A→C modification of nucleotide 302; G→C modification of nucleotide 304; T→C modification of nucleotide 307; A→G modification of nucleotide 310; A→C modification of nucleotide 313; T→G modification of nucleotide 316; A→C modification of nucleotide 319; T→C modification of nucleotide 325; T→C modification of nucleotide 334; T→C modification of nucleotide 337; T→G modification of nucleotide 340; T→C modification of nucleotide 343; T→C modification of nucleotide 346; A→G modification of nucleotide 349; A→C modification of nucleotide 352; A→C modification of nucleotide 353; A→C modification of nucleotide 355; A→G modification of nucleotide 358; A→G modification of nucleotide 361; C→G modification of nucleotide 367; T→C modification of nucleotide 382; A→T modification of nucleotide 383; G→C modification of nucleotide 384; A→C modification of nucleotide 391; A→T modification of nucleotide 392; G→C modification of nucleotide 393; T→C modification of nucleotide 394; A→C modification of nucleotide 397; A→G modification of nucleotide 403; A→G modification of nucleotide 406; A→G modification of nucleotide 409; T→C modification of nucleotide 412; T→C modification of nucleotide 418; A→G modification of nucleotide 421; T→C modification of nucleotide 424; C→G modification of nucleotide 430; T→C modification of nucleotide 433; A→G modification of nucleotide 436; A→G modification of nucleotide 442; T→C modification of nucleotide 451; A→C modification of nucleotide 452; A→C modification of nucleotide 454; T→C modification of nucleotide 466; T→C modification of nucleotide 469; A→G modification of nucleotide 472; A→C modification of nucleotide 475; A→T modification of nucleotide 479; G→C modification of nucleotide 480; A→C modification of nucleotide 490; A→C modification of nucleotide 493; A→G modification of nucleotide 496; T→C modification of nucleotide 499; T→C modification of nucleotide 505; A→T modification of nucleotide 509; G→C modification of nucleotide 510; T→C modification of nucleotide 511; A→G modification of nucleotide 520; T→C modification of nucleotide 523; A→T modification of nucleotide 533; G→C modification of nucleotide 534; T→C modification of nucleotide 535; A→C modification of nucleotide 544; A→C modification of nucleotide 547; A→G modification of nucleotide 553; A→T modification of nucleotide 557; G→C modification of nucleotide 558; T→C modification of nucleotide 562; T→C modification of nucleotide 563; T→C modification of nucleotide 568; G→C modification of nucleotide 571; A→T modification of nucleotide 572; G→C modification of nucleotide 573; T→C modification of nucleotide 577; G→C modification of nucleotide 580; T→C modification of nucleotide 583; A→G modification of nucleotide 586; A→G modification of nucleotide 592; A→G modification of nucleotide 598; T→C modification of nucleotide 607; A→C modification of nucleotide 610; T→C modification of nucleotide 613; T→C modification of nucleotide 616; A→C modification of nucleotide 619; G→C modification of nucleotide 622; A→G modification of nucleotide 625; A→G modification of nucleotide 628; G→C modification of nucleotide 631; A→C modification of nucleotide 634; T→C modification of nucleotide 643; A→C modification of nucleotide 649; A→T modification of nucleotide 650; G→C modification of nucleotide 651; T→C modification of nucleotide 655; A→C modification of nucleotide 658; A→G modification of nucleotide 664; A→G modification of nucleotide 673; C→G modification of nucleotide 676; T→C modification of nucleotide 679; T→C modification of nucleotide 685; T→C modification of nucleotide 688; T→C modification of nucleotide 694; A→C modification of nucleotide 697; A→G modification of nucleotide 700; T→G modification of nucleotide 703; A→G modification of nucleotide 706; T→C modification of nucleotide 721; G→C modification of nucleotide 724; A→T modification of nucleotide 728; G→C modification of nucleotide 729; T→C modification of nucleotide 730; T→C modification of nucleotide 733; A→C modification of nucleotide 736; T→C modification of nucleotide 739; C→G modification of nucleotide 742; T→C modification of nucleotide 748; T→C modification of nucleotide 751; A→C modification of nucleotide 754; A→G modification of nucleotide 757; A→C modification of nucleotide 760; T→C modification of nucleotide 769; T→C modification of nucleotide 772; C→G modification of nucleotide 778; T→C modification of nucleotide 784; T→C modification of nucleotide 787; A→C modification of nucleotide 783; A→C modification of nucleotide 786; G→C modification of nucleotide 802; A→G modification of nucleotide 808; T→C modification of nucleotide 812; T→C modification of nucleotide 817; A→G modification of nucleotide 820; A→C modification of nucleotide 821; A→G modification of nucleotide 823; T→C modification of nucleotide 826; T→C modification of nucleotide 841; T→C modification of nucleotide 844; T→C modification of nucleotide 847; T→C modification of nucleotide 850; T→C modification of nucleotide 853; T→C modification of nucleotide 862; T→C modification of nucleotide 868; G→C modification of nucleotide 877; A→C modification of nucleotide 878; G→C modification of nucleotide 880; A→C modification of nucleotide 883; T→C modification of nucleotide 892; T→C modification of nucleotide 895; T→G modification of nucleotide 898; T→C modification of nucleotide 904; T→C modification of nucleotide 925; A→C modification of nucleotide 928; T→C modification of nucleotide 931; A→G modification of nucleotide 934; A→C modification of nucleotide 937; T→C modification of nucleotide 943; T→C modification of nucleotide 949; T→C modification of nucleotide 952; T→C modification of nucleotide 958; T→C modification of nucleotide 964; T→C modification of nucleotide 967; T→G modification of nucleotide 970; T→C modification of nucleotide 973; T→C modification of nucleotide 974; A→G modification of nucleotide 979; A→C modification of nucleotide 982; T→C modification of nucleotide 985; A→G modification of nucleotide 988; A→G modification of nucleotide 997; T→C modification of nucleotide 1000; A→G modification of nucleotide 1003; A→C modification of nucleotide 1006; A→G modification of nucleotide 1009; A→C modification of nucleotide 1016; G→C modification of nucleotide 1018; T→C modification of nucleotide 1024; C→G modification of nucleotide 1027; T→C modification of nucleotide 1030; T→C modification of nucleotide 1036; T→C modification of nucleotide 1042; A→C modification of nucleotide 1045; T→C modification of nucleotide 1048; A→C modification of nucleotide 1051; T→C modification of nucleotide 1054; T→G modification of nucleotide 1060; T→C modification of nucleotide 1066; T→C modification of nucleotide 1069; T→C modification of nucleotide 1075; T→C modification of nucleotide 1078; T→C modification of nucleotide 1087; T→C modification of nucleotide 1096; T→C modification of nucleotide 1099; A→G modification of nucleotide 1102; T→C modification of nucleotide 1105; A→G modification of nucleotide 1123; A→C modification of nucleotide 1126; A→G modification of nucleotide 1129; T→C modification of nucleotide 1132; A→C modification of nucleotide 1141; T→C modification of nucleotide 1144; A→C modification of nucleotide 1147; T→C modification of nucleotide 1150; G→C modification of nucleotide 1153; T→C modification of nucleotide 1156; A→G modification of nucleotide 1159; A→C modification of nucleotide 1162; T→C modification of nucleotide 1171; T→C modification of nucleotide 1174; T→C modification of nucleotide 1180; T→C modification of nucleotide 1183; A→C modification of nucleotide 1186; A→C modification of nucleotide 1195; T→G modification of nucleotide 1198; A→G modification of nucleotide 1213; T→C modification of nucleotide 1216; A→C modification of nucleotide 1219; T→C modification of nucleotide 1225; T→C modification of nucleotide 1228; A→C modification of nucleotide 1234; T→C modification of nucleotide 1237; A→C modification of nucleotide 1240; A→G modification of nucleotide 1243; T→C modification of nucleotide 1246; T→C modification of nucleotide 1249; T→C modification of nucleotide 1252; T→C modification of nucleotide 1255; G→C modification of nucleotide 1258; T→C modification of nucleotide 1261; A→G modification of nucleotide 1267; T→C modification of nucleotide 1270; T→C modification of nucleotide 1273; T→C modification of nucleotide 1283; A→G modification of nucleotide 1285; A→G modification of nucleotide 1291; A→C modification of nucleotide 1294; A→C modification of nucleotide 1295; A→C modification of nucleotide 1297; T→C modification of nucleotide 1300; A→C modification of nucleotide 1303; A→G modification of nucleotide 1306; A→C modification of nucleotide 1309; T→C modification of nucleotide 1312; A→C modification of nucleotide 1315; A→G modification of nucleotide 1336; T→C modification of nucleotide 1340; A→G modification of nucleotide 1342; T→C modification of nucleotide 1345; T→C modification of nucleotide 1348; T→C modification of nucleotide 1351; A→G modification of nucleotide 1363; A→C modification of nucleotide 1366; T→C modification of nucleotide 1369; A→C modification of nucleotide 1372; A→T modification of nucleotide 1373; G→C modification of nucleotide 1374; T→C modification of nucleotide 1375; T→C modification of nucleotide 1384; T→C modification of nucleotide 1390; A→C modification of nucleotide 1399; T→C modification of nucleotide 1402; A→C modification of nucleotide 1405; T→C modification of nucleotide 1408; T→C modification of nucleotide 1411; T→C modification of nucleotide 1414; T→C modification of nucleotide 1417; T→C modification of nucleotide 1420; A→C modification of nucleotide 1423; G→C modification of nucleotide 1438; A→C modification of nucleotide 1441; T→C modification of nucleotide 1444; A→C modification of nucleotide 1447; C→G modification of nucleotide 1450; T→C modification of nucleotide 1453; T→C modification of nucleotide 1456; T→C modification of nucleotide 1462; T→C modification of nucleotide 1465; T→C modification of nucleotide 1471; T→C modification of nucleotide 1474; T→C modification of nucleotide 1477; T→C modification of nucleotide 1480; T→C modification of nucleotide 1483; A→G modification of nucleotide 1486; T→C modification of nucleotide 1489; T→C modification of nucleotide 1492; A→C modification of nucleotide 1498; T→C modification of nucleotide 1501; A→C modification of nucleotide 1504; A→C modification of nucleotide 1507; C→G modification of nucleotide 1510; T→C modification of nucleotide 1513; T→C modification of nucleotide 1514; A→G modification of nucleotide 1516; T→C modification of nucleotide 1522; A→G modification of nucleotide 1528; A→C modification of nucleotide 1531; T→C modification of nucleotide 1534; T→C modification of nucleotide 1537; A→G modification of nucleotide 1543; G→C modification of nucleotide 1546; A→G modification of nucleotide 1549; A→G modification of nucleotide 1552; T→C modification of nucleotide 1553; A→C modification of nucleotide 1558; T→G modification of nucleotide 1561; A→G modification of nucleotide 1564; T→C modification of nucleotide 1567; G→C modification of nucleotide 1570; T→C modification of nucleotide 1573; A→C modification of nucleotide 1576; A→C modification of nucleotide 1579; A→C modification of nucleotide 1582; A→C modification of nucleotide 1585; A→C modification of nucleotide 1594; T→G modification of nucleotide 1603; A→T modification of nucleotide 1604; G→C modification of nucleotide 1605; T→C modification of nucleotide 1606; T→C modification of nucleotide 1607; A→C modification of nucleotide 1610; A→C modification of nucleotide 1612; A→C modification of nucleotide 1616; A→C modification of nucleotide 1618; T→C modification of nucleotide 1621; A→G modification of nucleotide 1624; T→C modification of nucleotide 1627; A→C modification of nucleotide 1636; A→C modification of nucleotide 1639; A→C modification of nucleotide 1642; T→C modification of nucleotide 1643; A→C modification of nucleotide 1648; A→T modification of nucleotide 1655; G→C modification of nucleotide 1656; T→C modification of nucleotide 1657; T→G modification of nucleotide 1663; T→G modification of nucleotide 1666; T→C modification of nucleotide 1669; A→C modification of nucleotide 1672; A→C modification of nucleotide 1675; T→C modification of nucleotide 1681; T→C modification of nucleotide 1687; T→C modification of nucleotide 1690; A→C modification of nucleotide 1693; T→C modification of nucleotide 1697; A→G modification of nucleotide 1702; T→C modification of nucleotide 1705; T→C modification of nucleotide 1708; A→G modification of nucleotide 1711; T→C modification of nucleotide 1714; T→G modification of nucleotide 1720; A→C modification of nucleotide 1723; T→C modification of nucleotide 1726; T→C modification of nucleotide 1729; T→C modification of nucleotide 1732; C→G modification of nucleotide 1735; A→C modification of nucleotide 1741; A→C modification of nucleotide 1742; A→C modification of nucleotide 1744; A→C modification of nucleotide 1747; T→C modification of nucleotide 1750; A→G modification of nucleotide 1756; A→G modification of nucleotide 1762; T→G modification of nucleotide 1768; C→G modification of nucleotide 1771; T→C modification of nucleotide 1774; T→G modification of nucleotide 1777; A→G modification of nucleotide 1789; T→C modification of nucleotide 1792; T→C modification of nucleotide 1798; A→G modification of nucleotide 1801; A→C modification of nucleotide 1804; A→C modification of nucleotide 1807; T→C modification of nucleotide 1810; T→G modification of nucleotide 1816; T→C modification of nucleotide 1817; A→G modification of nucleotide 1819; T→G modification of nucleotide 1828; A→C modification of nucleotide 1832; G→C modification of nucleotide 1834; T→C modification of nucleotide 1837; T→C modification of nucleotide 1840; C→G modification of nucleotide 1843; T→C modification of nucleotide 1849; T→C modification of nucleotide 1852; T→C modification of nucleotide 1855; T→G modification of nucleotide 1858; G→C modification of nucleotide 1860; T→C modification of nucleotide 1861; G→C modification of nucleotide 1863; T→C modification of nucleotide 1866; T→C modification of nucleotide 1870; T→C modification of nucleotide 1871; A→G modification of nucleotide 1873; T→C modification of nucleotide 1876; T→C modification of nucleotide 1879; T→C modification of nucleotide 1882; A→C modification of nucleotide 1885; A→C modification of nucleotide 1891; T→C modification of nucleotide 1894; T→C modification of nucleotide 1897; A→G modification of nucleotide 1900; A→T modification of nucleotide 1907; G→C modification of nucleotide 1908; T→C modification of nucleotide 1909; A→T modification of nucleotide 1913; G→C modification of nucleotide 1914; T→C modification of nucleotide 1915; T→G modification of nucleotide 1918; T→C modification of nucleotide 1921; T→C modification of nucleotide 1936; A→C modification of nucleotide 1939; T→C modification of nucleotide 1940; T→C modification of nucleotide 1951; T→C modification of nucleotide 1954; A→G modification of nucleotide 1960; T→C modification of nucleotide 1961; A→G modification of nucleotide 1963; A→C modification of nucleotide 1966; A→C modification of nucleotide 1972; T→C modification of nucleotide 1975; C→G modification of nucleotide 1978; T→C modification of nucleotide 1981; A→C modification of nucleotide 1990; T→C modification of nucleotide 1993; A→G modification of nucleotide 1999; A→T modification of nucleotide 2003; G→C modification of nucleotide 2004; T→C modification of nucleotide 2011; T→C modification of nucleotide 2014; A→C modification of nucleotide 2017; G→C modification of nucleotide 2020; T→C modification of nucleotide 2029; T→C modification of nucleotide 2035; T→C modification of nucleotide 2041; A→G modification of nucleotide 2044; A→C modification of nucleotide 2047; T→C modification of nucleotide 2050; T→C modification of nucleotide 2053; G→C modification of nucleotide 2056; T→C modification of nucleotide 2059; A→G modification of nucleotide 2062; T→C modification of nucleotide 2068; A→C modification of nucleotide 2071; A→C modification of nucleotide 2074; A→C modification of nucleotide 2077; A→C modification of nucleotide 2080; T→C modification of nucleotide 2083; G→C modification of nucleotide 2089; T→C modification of nucleotide 2092; T→C modification of nucleotide 2095; T→C modification of nucleotide 2098; A→C modification of nucleotide 2104; T→G modification of nucleotide 2107; T→G modification of nucleotide 2110; T→C modification of nucleotide 2113; A→G modification of nucleotide 2119; T→C modification of nucleotide 2122; A→G modification of nucleotide 2125; A→C modification of nucleotide 2129; A→C modification of nucleotide 2131; T→G modification of nucleotide 2137; T→G modification of nucleotide 2140; T→C modification of nucleotide 2143; C→G modification of nucleotide 2146; T→C modification of nucleotide 2149; T→G modification of nucleotide 2152; T→C modification of nucleotide 2155; T→C modification of nucleotide 2158; T→C modification of nucleotide 2161; A→C modification of nucleotide 2164; T→C modification of nucleotide 2167; T→C modification of nucleotide 2173; A→C modification of nucleotide 2176; T→C modification of nucleotide 2179; T→C modification of nucleotide 2182; T→C modification of nucleotide 2185; T→C modification of nucleotide 2188; T→C modification of nucleotide 2191; T→C modification of nucleotide 2194; C→G modification of nucleotide 2197; A→C modification of nucleotide 2200; A→G modification of nucleotide 2203; A→C modification of nucleotide 2206; T→C modification of nucleotide 2209; T→C modification of nucleotide 2212; A→C modification of nucleotide 2215; A→G modification of nucleotide 2218; A→C modification of nucleotide 2224; A→G modification of nucleotide 2230; T→C modification of nucleotide 2233; T→C modification of nucleotide 2236; T→C modification of nucleotide 2237; A→G modification of nucleotide 2239; A→C modification of nucleotide 2242; T→C modification of nucleotide 2245; A→G modification of nucleotide 2251; A→C modification of nucleotide 2254; A→C modification of nucleotide 2262; and deletion of nucleotides 2263-5834;

›DETAILED DESCRIPTION · 23 of 32

d) a sequence encoding Cyclin-Dependent Kinase Inhibitor 1A (P21) and comprising one or more of the following modifications relative to SEQ ID NO: 4:

deletion of nucleotides 1-134; T→C modification of nucleotide 143; T→C modification of nucleotide 146; T→C modification of nucleotide 149; T→C modification of nucleotide 152; C→G modification of nucleotide 155; A→C modification of nucleotide 158; T→C modification of nucleotide 161; T→G modification of nucleotide 164; G→C modification of nucleotide 167; A→C modification of nucleotide 171; G→C modification of nucleotide 173; A→T modification of nucleotide 174; G→C modification of nucleotide 175; A→G modification of nucleotide 179; T→C modification of nucleotide 188; T→C modification of nucleotide 191; C→G modification of nucleotide 194; T→C modification of nucleotide 200; A→T modification of nucleotide 210; G→C modification of nucleotide 211; T→C modification of nucleotide 212; T→C modification of nucleotide 219; T→C modification of nucleotide 227; T→C modification of nucleotide 230; T→C modification of nucleotide 236; G→C modification of nucleotide 239; C→G modification of nucleotide 242; G→C modification of nucleotide 248; T→C modification of nucleotide 254; C→G modification of nucleotide 257; A→C modification of nucleotide 269; A→G modification of nucleotide 272; T→C modification of nucleotide 284; C→G modification of nucleotide 293; G→C modification of nucleotide 296; G→C modification of nucleotide 302; G→C modification of nucleotide 305; C→G modification of nucleotide 323; T→G modification of nucleotide 335; G→C modification of nucleotide 338; A→T modification of nucleotide 339; G→C modification of nucleotide 340; A→G modification of nucleotide 344; G→C modification of nucleotide 347; C→G modification of nucleotide 359; A→T modification of nucleotide 366; G→C modification of nucleotide 367; T→C modification of nucleotide 371; G→C modification of nucleotide 374; A→T modification of nucleotide 381; G→C modification of nucleotide 382; T→C modification of nucleotide 386; A→C modification of nucleotide 398; G→C modification of nucleotide 401; A→C modification of nucleotide 408; G→C modification of nucleotide 410; A→T modification of nucleotide 414; G→C modification of nucleotide 415; T→C modification of nucleotide 416; T→C modification of nucleotide 419; T→C modification of nucleotide 425; G→C modification of nucleotide 440; A→C modification of nucleotide 443; T→C modification of nucleotide 446; G→C modification of nucleotide 449; T→C modification of nucleotide 465; G→C modification of nucleotide 470; T→C modification of nucleotide 476; T→C modification of nucleotide 482; T→C modification of nucleotide 491; G→C modification of nucleotide 497; T→C modification of nucleotide 500; A→G modification of nucleotide 503; T→C modification of nucleotide 506; G→C modification of nucleotide 512; T→C modification of nucleotide 515; G→C modification of nucleotide 518; A→C modification of nucleotide 524; A→C modification of nucleotide 527; T→C modification of nucleotide 530; A→C modification of nucleotide 539; A→G modification of nucleotide 542; G→C modification of nucleotide 545; A→C modification of nucleotide 546; G→C modification of nucleotide 548; A→G modification of nucleotide 555; G→C modification of nucleotide 556; A→C modification of nucleotide 563; T→C modification of nucleotide 566; T→C modification of nucleotide 572; A→C modification of nucleotide 585; A→C modification of nucleotide 587; T→C modification of nucleotide 588; C→G modification of nucleotide 593; A→C modification of nucleotide 603; A→C modification of nucleotide 605; A→G modification of nucleotide 608; and deletion of nucleotides 613-1943;

e) a sequence encoding Vascular Endothelial Growth Factor A (VEGFA) and comprising one or more of the following modifications relative to SEQ ID NO: 5:

deletion of nucleotides 1-1036; T→C modification of nucleotide 1045; T→C modification of nucleotide 1054; T→C modification of nucleotide 1063; A→T modification of nucleotide 1067; G→C modification of nucleotide 1068; T→G modification of nucleotide 1072; T→C modification of nucleotide 1076; C→G modification of nucleotide 1084; C→G modification of nucleotide 1090; T→C modification of nucleotide 1096; T→C modification of nucleotide 1114; A→C modification of nucleotide 1117; A→C modification of nucleotide 1126; A→G modification of nucleotide 1129; A→C modification of nucleotide 1132; A→C modification of nucleotide 1135; G→C modification of nucleotide 1138; T→C modification of nucleotide 1144; T→C modification of nucleotide 1147; A→G modification of nucleotide 1153; T→C modification of nucleotide 1171; C→G modification of nucleotide 1174; T→C modification of nucleotide 1177; C→G modification of nucleotide 1183; A→T modification of nucleotide 1184; G→C modification of nucleotide 1185; T→C modification of nucleotide 1195; A→C modification of nucleotide 1198; T→C modification of nucleotide 1234; T→C modification of nucleotide 1237; A→C modification of nucleotide 1261; T→C modification of nucleotide 1267; A→C modification of nucleotide 1282; G→C modification of nucleotide 1288; T→C modification of nucleotide 1300; T→C modification of nucleotide 1318; T→C modification of nucleotide 1327; T→C modification of nucleotide 1354; G→C modification of nucleotide 1360; A→G modification of nucleotide 1366; T→C modification of nucleotide 1369; A→G modification of nucleotide 1375; A→C modification of nucleotide 1387; A→C modification of nucleotide 1390; A→T modification of nucleotide 1397; G→C modification of nucleotide 1398; A→G modification of nucleotide 1405; A→G modification of nucleotide 1417; T→C modification of nucleotide 1420; A→G modification of nucleotide 1423; A→C modification of nucleotide 1427; A→C modification of nucleotide 1429; A→C modification of nucleotide 1432; A→G modification of nucleotide 1438; T→C modification of nucleotide 1441; A→C modification of nucleotide 1442; A→C modification of nucleotide 1444; A→C modification of nucleotide 1447; A→C modification of nucleotide 1448; A→C modification of nucleotide 1450; A→G modification of nucleotide 1453; A→G modification of nucleotide 1456; T→C modification of nucleotide 1459; T→C modification of nucleotide 1465; G→C modification of nucleotide 1468; T→C modification of nucleotide 1471; A→C modification of nucleotide 1477; G→C modification of nucleotide 1483; A→C modification of nucleotide 1484; A→C modification of nucleotide 1486; T→C modification of nucleotide 1492; T→C modification of nucleotide 1493; T→C modification of nucleotide 1498; A→G modification of nucleotide 1501; A→G modification of nucleotide 1504; T→C modification of nucleotide 1507; G→C modification of nucleotide 1510; G→C modification of nucleotide 1516; T→C modification of nucleotide 1519; A→G modification of nucleotide 1522; T→C modification of nucleotide 1525; A→G modification of nucleotide 1534; A→C modification of nucleotide 1540; G→C modification of nucleotide 1546; T→C modification of nucleotide 1549; and deletion of nucleotides 1556-3502;

›DETAILED DESCRIPTION · 24 of 32

f) a sequence encoding Insulin-like Growth Factor 1 (Igf1) and comprising one or more of the following modifications relative to SEQ ID NO: 6:

deletion of nucleotides 1-298; G→C modification of nucleotide 304; A→G modification of nucleotide 307; A→T modification of nucleotide 311; G→C modification of nucleotide 312; A→T modification of nucleotide 314; G→C modification of nucleotide 315; T→G modification of nucleotide 319; A→C modification of nucleotide 322; T→C modification of nucleotide 325; A→G modification of nucleotide 328; T→C modification of nucleotide 329; A→G modification of nucleotide 331; T→C modification of nucleotide 334; C→G modification of nucleotide 346; T→C modification of nucleotide 349; T→C modification of nucleotide 356; A→C modification of nucleotide 364; A→C modification of nucleotide 370; G→C modification of nucleotide 382; T→C modification of nucleotide 385; A→C modification of nucleotide 388; C→G modification of nucleotide 394; G→C modification of nucleotide 406; C→G modification of nucleotide 409; T→C modification of nucleotide 413; C→G modification of nucleotide 418; A→T modification of nucleotide 428; G→C modification of nucleotide 429; A→C modification of nucleotide 439; T→C modification of nucleotide 442; A→C modification of nucleotide 445; A→C modification of nucleotide 448; T→G modification of nucleotide 457; G→C modification of nucleotide 463; T→C modification of nucleotide 466; T→C modification of nucleotide 478; T→C modification of nucleotide 481; T→G modification of nucleotide 484; T→C modification of nucleotide 496; A→C modification of nucleotide 499; G→C modification of nucleotide 502; A→C modification of nucleotide 503; G→C modification of nucleotide 505; T→C modification of nucleotide 511; A→C modification of nucleotide 529; T→C modification of nucleotide 535; A→T modification of nucleotide 542; G→C modification of nucleotide 543; T→C modification of nucleotide 547; G→C modification of nucleotide 550; A→C modification of nucleotide 551; G→C modification of nucleotide 553; A→C modification of nucleotide 556; T→C modification of nucleotide 559; A→C modification of nucleotide 565; T→C modification of nucleotide 571; T→C modification of nucleotide 577; T→C modification of nucleotide 583; G→C modification of nucleotide 592; A→T modification of nucleotide 593; G→C modification of nucleotide 594; T→C modification of nucleotide 598; T→C modification of nucleotide 601; A→C modification of nucleotide 605; G→C modification of nucleotide 607; A→C modification of nucleotide 608; A→C modification of nucleotide 610; T→C modification of nucleotide 625; A→C modification of nucleotide 631; T→C modification of nucleotide 640; A→C modification of nucleotide 643; A→G modification of nucleotide 646; A→C modification of nucleotide 649; T→C modification of nucleotide 658; T→C modification of nucleotide 664; T→C modification of nucleotide 679; T→C modification of nucleotide 694; A→G modification of nucleotide 703; A→G modification of nucleotide 706; T→C modification of nucleotide 709; T→C modification of nucleotide 710; A→C modification of nucleotide 721; A→T modification of nucleotide 722; G→C modification of nucleotide 723; T→C modification of nucleotide 724; A→C modification of nucleotide 725; A→C modification of nucleotide 727; A→C modification of nucleotide 730; A→T modification of nucleotide 731; G→C modification of nucleotide 732; T→C modification of nucleotide 733; A→C modification of nucleotide 736; A→C modification of nucleotide 739; A→C modification of nucleotide 752; A→C modification of nucleotide 754; and deletion of nucleotides 760-7073;

g) a sequence encoding Insulin-like Growth Factor 1 (Igf1) and comprising one or more of the following modifications relative to SEQ ID NO: 6:

deletion of nucleotides 1-442; insertion of SEQ ID NO: 18; A→C modification of nucleotide 445; A→C modification of nucleotide 448; T→G modification of nucleotide 457; G→C modification of nucleotide 463; T→C modification of nucleotide 466; T→C modification of nucleotide 478; T→C modification of nucleotide 481; T→G modification of nucleotide 484; T→C modification of nucleotide 496; A→C modification of nucleotide 499; G→C modification of nucleotide 502; A→C modification of nucleotide 503; G→C modification of nucleotide 505; T→C modification of nucleotide 511; A→C modification of nucleotide 529; T→C modification of nucleotide 535; A→T modification of nucleotide 542; G→C modification of nucleotide 543; T→C modification of nucleotide 547; G→C modification of nucleotide 550; A→C modification of nucleotide 551; G→C modification of nucleotide 553; A→C modification of nucleotide 556; T→C modification of nucleotide 559; A→C modification of nucleotide 565; T→C modification of nucleotide 571; T→C modification of nucleotide 577; T→C modification of nucleotide 583; G→C modification of nucleotide 592; A→T modification of nucleotide 593; G→C modification of nucleotide 594; T→C modification of nucleotide 598; T→C modification of nucleotide 601; A→C modification of nucleotide 605; G→C modification of nucleotide 607; A→C modification of nucleotide 608; A→C modification of nucleotide 610; T→C modification of nucleotide 625; A→C modification of nucleotide 631; T→C modification of nucleotide 640; A→C modification of nucleotide 643; A→G modification of nucleotide 646; A→C modification of nucleotide 649; T→C modification of nucleotide 658; T→C modification of nucleotide 664; T→C modification of nucleotide 679; T→C modification of nucleotide 694; A→G modification of nucleotide 703; A→G modification of nucleotide 706; T→C modification of nucleotide 709; T→C modification of nucleotide 710; A→C modification of nucleotide 721; A→T modification of nucleotide 722; G→C modification of nucleotide 723; T→C modification of nucleotide 724; A→C modification of nucleotide 725; A→C modification of nucleotide 727; A→C modification of nucleotide 730; A→T modification of nucleotide 731; G→C modification of nucleotide 732; T→C modification of nucleotide 733; A→C modification of nucleotide 736; A→C modification of nucleotide 739; A→C modification of nucleotide 752; A→C modification of nucleotide 754; and deletion of nucleotides 760-7073;

›DETAILED DESCRIPTION · 25 of 32

h) a sequence encoding Epidermal Growth Factor1 (EGF) and comprising one or more of the following modifications relative to SEQ ID NO: 2 deletion of nucleotides 1-3369; insertion of SEQ ID NO: 19; T→C modification of nucleotide 3375; A→G modification of nucleotide 3378; T→C modification of nucleotide 3381; T→C modification of nucleotide 3396; G→C modification of nucleotide 3399; C→G modification of nucleotide 3408; T→C modification of nucleotide 3411; T→C modification of nucleotide 3414; T→C modification of nucleotide 3417; T→C modification of nucleotide 3429; T→C modification of nucleotide 3432; A→G modification of nucleotide 3435; A→C modification of nucleotide 3438; T→C modification of nucleotide 3439; T→C modification of nucleotide 3450; A→C modification of nucleotide 3453; T→C modification of nucleotide 3462; T→G modification of nucleotide 3465; T→G modification of nucleotide 3468; G→C modification of nucleotide 3480; A→G modification of nucleotide 3486; T→C modification of nucleotide 3489; A→C modification of nucleotide 3498; A→G modification of nucleotide 3516; and deletion of nucleotides 3523-6388; i) a sequence encoding signal transducer and activator of transcription 5B (Stat5b) and comprising one or more of the following modifications relative to SEQ ID NO: 7:

deletion of nucleotides 1-532; insertion of SEQ ID NO: 20; T→C modification of nucleotide 535; A→C modification of nucleotide 544; T→C modification of nucleotide 550; C→G modification of nucleotide 559; T→C modification of nucleotide 568; T→G modification of nucleotide 574; T→C modification of nucleotide 590; T→C modification of nucleotide 604; A→C modification of nucleotide 622; T→C modification of nucleotide 625; T→C modification of nucleotide 628; T→C modification of nucleotide 629; A→G modification of nucleotide 631; A→C modification of nucleotide 634; A→G modification of nucleotide 646; A→T modification of nucleotide 647; G→C modification of nucleotide 648; A→G modification of nucleotide 652; A→C modification of nucleotide 664; A→C modification of nucleotide 667; T→C modification of nucleotide 670; T→G modification of nucleotide 673; T→C modification of nucleotide 676; T→C modification of nucleotide 679; A→C modification of nucleotide 682; T→C modification of nucleotide 694; C→G modification of nucleotide 709; G→C modification of nucleotide 745; G→C modification of nucleotide 760; A→G modification of nucleotide 763; T→C modification of nucleotide 766; G→C modification of nucleotide 769; T→C modification of nucleotide 772; T→C modification of nucleotide 773; G→C modification of nucleotide 793; T→C modification of nucleotide 799; A→C modification of nucleotide 805; C→G modification of nucleotide 811; A→T modification of nucleotide 815; G→C modification of nucleotide 816; G→C modification of nucleotide 820; T→G modification of nucleotide 847; T→C modification of nucleotide 853; G→C modification of nucleotide 859; T→C modification of nucleotide 865; A→G modification of nucleotide 877; A→C modification of nucleotide 881; G→C modification of nucleotide 883; T→G modification of nucleotide 889; A→G modification of nucleotide 895; A→T modification of nucleotide 908; G→C modification of nucleotide 909; T→C modification of nucleotide 913; A→C modification of nucleotide 916; T→C modification of nucleotide 919; A→C modification of nucleotide 922; A→T modification of nucleotide 923; G→C modification of nucleotide 924; T→C modification of nucleotide 925; T→G modification of nucleotide 928; T→C modification of nucleotide 931; T→G modification of nucleotide 955; A→G modification of nucleotide 967; G→C modification of nucleotide 970; T→C modification of nucleotide 973; A→C modification of nucleotide 994; G→C modification of nucleotide 1003; A→G modification of nucleotide 1036; A→T modification of nucleotide 1064; G→C modification of nucleotide 1065; G→C modification of nucleotide 1072; A→G modification of nucleotide 1078; T→C modification of nucleotide 1081; T→C modification of nucleotide 1087; A→C modification of nucleotide 1099; A→T modification of nucleotide 1124; G→C modification of nucleotide 1125; A→C modification of nucleotide 1127; G→C modification of nucleotide 1129; G→C modification of nucleotide 1135; C→G modification of nucleotide 1141; A→G modification of nucleotide 1153; A→C modification of nucleotide 1180; A→C modification of nucleotide 1186; A→C modification of nucleotide 1192; A→C modification of nucleotide 1207; T→C modification of nucleotide 1219; G→C modification of nucleotide 1252; G→C modification of nucleotide 1300; A→C modification of nucleotide 1301; A→C modification of nucleotide 1303; G→C modification of nucleotide 1318; G→C modification of nucleotide 1324; T→C modification of nucleotide 1327; A→T modification of nucleotide 1340; G→C modification of nucleotide 1341; T→C modification of nucleotide 1366; G→C modification of nucleotide 1399; A→C modification of nucleotide 1412; G→C modification of nucleotide 1414; T→C modification of nucleotide 1417; T→C modification of nucleotide 1424; A→C modification of nucleotide 1447; T→C modification of nucleotide 1471; C→G modification of nucleotide 1477; G→C modification of nucleotide 1492; A→C modification of nucleotide 1504; C→G modification of nucleotide 1513; A→T modification of nucleotide 1517; G→C modification of nucleotide 1518; G→C modification of nucleotide 1522; T→C modification of nucleotide 1543; T→C modification of nucleotide 1546; C→G modification of nucleotide 1552; T→C modification of nucleotide 1573; A→C modification of nucleotide 1576; G→C modification of nucleotide 1600; G→C modification of nucleotide 1603; T→C modification of nucleotide 1612; G→C modification of nucleotide 1630; G→C modification of nucleotide 1642; A→T modification of nucleotide 1652; G→C modification of nucleotide 1653; C→G modification of nucleotide 1678; T→C modification of nucleotide 1684; T→C modification of nucleotide 1699; T→C modification of nucleotide 1702; A→T modification of nucleotide 1706; G→C modification of nucleotide 1707; T→C modification of nucleotide 1729; C→G modification of nucleotide 1735; T→C modification of nucleotide 1756; A→C modification of nucleotide 1762; C→G modification of nucleotide 1765; A→T modification of nucleotide 1766; G→C modification of nucleotide 1767; A→C modification of nucleotide 1778; A→C modification of nucleotide 1780; A→G modification of nucleotide 1795; A→C modification of nucleotide 1798; A→C modification of nucleotide 1805; T→C modification of nucleotide 1810; T→C modification of nucleotide 1819; G→C modification of nucleotide 1822; A→C modification of nucleotide 1825; A→C modification of nucleotide 1831; A→G modification of nucleotide 1834; G→C modification of nucleotide 1837; A→G modification of nucleotide 1840; G→C modification of nucleotide 1852; T→C modification of nucleotide 1861; A→C modification of nucleotide 1867; A→T modification of nucleotide 1874; G→C modification of nucleotide 1875; C→G modification of nucleotide 1879; T→C modification of nucleotide 1882; A→C modification of nucleotide 1885; C→G modification of nucleotide 1897; T→C modification of nucleotide 1900; A→G modification of nucleotide 1903; C→G modification of nucleotide 1906; T→C modification of nucleotide 1913; G→C modification of nucleotide 1918; C→G modification of nucleotide 1921; G→C modification of nucleotide 1924; T→C modification of nucleotide 1936; T→G modification of nucleotide 1939; A→T modification of nucleotide 1946; G→C modification of nucleotide 1947; T→C modification of nucleotide 1960; A→C modification of nucleotide 1966; T→C modification of nucleotide 1972; C→G modification of nucleotide 1975; C→G modification of nucleotide 1978; T→C modification of nucleotide 1993; A→C modification of nucleotide 1996; T→C modification of nucleotide 2002; A→C modification of nucleotide 2006; G→C modification of nucleotide 2008; A→C modification of nucleotide 2014; T→C modification of nucleotide 2017; T→C modification of nucleotide 2026; G→C modification of nucleotide 2044; T→C modification of nucleotide 2053; A→G modification of nucleotide 2056; G→C modification of nucleotide 2059; C→G modification of nucleotide 2062; A→G modification of nucleotide 2071; T→C modification of nucleotide 2080; A→G modification of nucleotide 2083; A→G modification of nucleotide 2086; A→T modification of nucleotide 2090; G→C modification of nucleotide 2091; G→C modification of nucleotide 2098; T→C modification of nucleotide 2102; C→G modification of nucleotide 2119; A→C modification of nucleotide 2131; A→G modification of nucleotide 2137; A→T modification of nucleotide 2150; G→C modification of nucleotide 2151; A→T modification of nucleotide 2153; G→C modification of nucleotide 2154; C→G modification of nucleotide 2164; A→T modification of nucleotide 2177; G→C modification of nucleotide 2178; G→C modification of nucleotide 2209; T→C modification of nucleotide 2215; T→C modification of nucleotide 2216; A→C modification of nucleotide 2221; A→C modification of nucleotide 2224; G→C modification of nucleotide 2227; T→C modification of nucleotide 2230; T→C modification of nucleotide 2236; T→C modification of nucleotide 2251; T→C modification of nucleotide 2254; A→G modification of nucleotide 2266; A→G modification of nucleotide 2269; T→C modification of nucleotide 2270; A→G modification of nucleotide 2275; A→G modification of nucleotide 2278; T→C modification of nucleotide 2281; C→G modification of nucleotide 2284; T→C modification of nucleotide 2290; T→C modification of nucleotide 2299; T→C modification of nucleotide 2302; G→C modification of nucleotide 2305; T→C modification of nucleotide 2308; T→C modification of nucleotide 2317; A→G modification of nucleotide 2332; C→G modification of nucleotide 2350; A→C modification of nucleotide 2362; G→C modification of nucleotide 2368; A→T modification of nucleotide 2387; G→C modification of nucleotide 2388; G→C modification of nucleotide 2395; A→G modification of nucleotide 2398; G→C modification of nucleotide 2404; T→C modification of nucleotide 2416; T→C modification of nucleotide 2419; T→C modification of nucleotide 2428; T→C modification of nucleotide 2434; A→C modification of nucleotide 2441; A→C modification of nucleotide 2443; T→C modification of nucleotide 2449; A→C modification of nucleotide 2453; T→C modification of nucleotide 2455; T→C modification of nucleotide 2464; T→C modification of nucleotide 2467; T→C modification of nucleotide 2473; A→C modification of nucleotide 2474; A→C modification of nucleotide 2476; T→C modification of nucleotide 2485; G→C modification of nucleotide 2491; C→G modification of nucleotide 2497; T→C modification of nucleotide 2500; G→C modification of nucleotide 2512; T→C modification of nucleotide 2521; C→G modification of nucleotide 2527; A→C modification of nucleotide 2530; T→C modification of nucleotide 2533; T→C modification of nucleotide 2539; T→C modification of nucleotide 2542; T→C modification of nucleotide 2545; G→C modification of nucleotide 2548; A→C modification of nucleotide 2551; T→C modification of nucleotide 2557; A→G modification of nucleotide 2560; A→G modification of nucleotide 2563; T→C modification of nucleotide 2566; T→C modification of nucleotide 2569; A→C modification of nucleotide 2581; G→C modification of nucleotide 2584; C→G modification of nucleotide 2587; T→C modification of nucleotide 2593; A→C modification of nucleotide 2602; T→C modification of nucleotide 2605; G→C modification of nucleotide 2608; A→G modification of nucleotide 2611; A→C modification of nucleotide 2614; T→C modification of nucleotide 2617; A→C modification of nucleotide 2623; A→C modification of nucleotide 2635; C→G modification of nucleotide 2653; T→C modification of nucleotide 2662; A→C modification of nucleotide 2665; T→C modification of nucleotide 2668; A→C modification of nucleotide 2671; A→C modification of nucleotide 2677; T→C modification of nucleotide 2680; T→C modification of nucleotide 2683; G→C modification of nucleotide 2686; A→T modification of nucleotide 2687; G→C modification of nucleotide 2688; T→C modification of nucleotide 2689; T→C modification of nucleotide 2707; T→C modification of nucleotide 2713; T→C modification of nucleotide 2716; A→C modification of nucleotide 2722; C→G modification of nucleotide 2725; T→C modification of nucleotide 2734; T→C modification of nucleotide 2740; A→C modification of nucleotide 2758; G→C modification of nucleotide 2767; C→G modification of nucleotide 2776; T→G modification of nucleotide 2779; T→C modification of nucleotide 2782; T→C modification of nucleotide 2788; G→C modification of nucleotide 2791; T→C modification of nucleotide 2800; A→G modification of nucleotide 2806; G→C modification of nucleotide 2812; G→C modification of nucleotide 2824; G→C modification of nucleotide 2827; A→G modification of nucleotide 2836; C→G modification of nucleotide 2842; T→C modification of nucleotide 2843; A→G modification of nucleotide 2845; G→C modification of nucleotide 2851; A→T modification of nucleotide 2861; G→C modification of nucleotide 2862; T→C modification of nucleotide 2863; T→C modification of nucleotide 2875; A→C modification of nucleotide 2881; A→C modification of nucleotide 2887; and deletion of nucleotides 2888-5255;

›DETAILED DESCRIPTION · 26 of 32

j) a sequence encoding beta catenin (CTNNB1) and comprising one or more of the following modifications relative to SEQ ID NO: 17:

deletion of nucleotides 1-214; T→C modification of nucleotide 220; T→C modification of nucleotide 223; A→G modification of nucleotide 226; T→C modification of nucleotide 229; T→C modification of nucleotide 232; T→C modification of nucleotide 233; T→C modification of nucleotide 242; A→G modification of nucleotide 259; A→C modification of nucleotide 262; A→C modification of nucleotide 266; A→C modification of nucleotide 268; A→G modification of nucleotide 271; G→C modification of nucleotide 274; T→C modification of nucleotide 277; T→G modification of nucleotide 280; A→T modification of nucleotide 281; G→C modification of nucleotide 282; T→C modification of nucleotide 283; A→G modification of nucleotide 295; T→C modification of nucleotide 301; T→C modification of nucleotide 313; A→C modification of nucleotide 316; T→C modification of nucleotide 322; T→C modification of nucleotide 325; T→C modification of nucleotide 328; T→C modification of nucleotide 334; A→C modification of nucleotide 340; T→C modification of nucleotide 343; T→C modification of nucleotide 346; T→C modification of nucleotide 349; A→C modification of nucleotide 353; G→C modification of nucleotide 354; T→C modification of nucleotide 355; T→C modification of nucleotide 358; A→G modification of nucleotide 361; T→C modification of nucleotide 367; T→C modification of nucleotide 370; A→G modification of nucleotide 376; T→C modification of nucleotide 382; T→C modification of nucleotide 388; A→G modification of nucleotide 397; C→G modification of nucleotide 400; T→C modification of nucleotide 406; A→G modification of nucleotide 415; A→C modification of nucleotide 421; T→C modification of nucleotide 424; T→C modification of nucleotide 427; T→C modification of nucleotide 439; A→G modification of nucleotide 442; A→G modification of nucleotide 445; A→G modification of nucleotide 448; A→G modification of nucleotide 451; T→C modification of nucleotide 454; T→C modification of nucleotide 457; T→C modification of nucleotide 460; T→C modification of nucleotide 463; A→C modification of nucleotide 466; T→C modification of nucleotide 472; A→C modification of nucleotide 475; T→C modification of nucleotide 481; A→C modification of nucleotide 484; T→C modification of nucleotide 487; A→C modification of nucleotide 491; G→C modification of nucleotide 493; A→G modification of nucleotide 496; A→C modification of nucleotide 499; T→C modification of nucleotide 502; T→C modification of nucleotide 505; T→C modification of nucleotide 514; A→C modification of nucleotide 520; T→C modification of nucleotide 521; A→G modification of nucleotide 523; T→C modification of nucleotide 526; A→C modification of nucleotide 544; T→C modification of nucleotide 547; A→C modification of nucleotide 550; T→C modification of nucleotide 556; T→C modification of nucleotide 559; T→C modification of nucleotide 562; T→C modification of nucleotide 565; T→C modification of nucleotide 568; T→C modification of nucleotide 574; T→C modification of nucleotide 577; C→G modification of nucleotide 580; T→C modification of nucleotide 586; T→C modification of nucleotide 587; T→C modification of nucleotide 592; A→G modification of nucleotide 595; A→C modification of nucleotide 598; A→C modification of nucleotide 601; A→G modification of nucleotide 613; T→C modification of nucleotide 616; A→C modification of nucleotide 619; T→G modification of nucleotide 622; A→G modification of nucleotide 625; T→C modification of nucleotide 629; T→C modification of nucleotide 634; T→C modification of nucleotide 640; A→G modification of nucleotide 643; T→C modification of nucleotide 646; T→C modification of nucleotide 649; A→C modification of nucleotide 652; A→G modification of nucleotide 655; T→G modification of nucleotide 658; A→C modification of nucleotide 664; T→C modification of nucleotide 667; A→C modification of nucleotide 670; T→C modification of nucleotide 676; A→G modification of nucleotide 679; A→C modification of nucleotide 685; A→G modification of nucleotide 688; A→G modification of nucleotide 694; T→C modification of nucleotide 697; T→G modification of nucleotide 718; T→C modification of nucleotide 721; T→C modification of nucleotide 727; A→C modification of nucleotide 730; T→G modification of nucleotide 733; C→G modification of nucleotide 739; T→C modification of nucleotide 742; T→G modification of nucleotide 748; T→C modification of nucleotide 751; A→G modification of nucleotide 754; A→G modification of nucleotide 760; T→C modification of nucleotide 763; A→C modification of nucleotide 767; A→C modification of nucleotide 769; T→C modification of nucleotide 775; T→C modification of nucleotide 784; T→C modification of nucleotide 787; T→C modification of nucleotide 790; T→C modification of nucleotide 802; T→C modification of nucleotide 805; T→C modification of nucleotide 808; A→G modification of nucleotide 811; T→C modification of nucleotide 814; T→C modification of nucleotide 826; A→C modification of nucleotide 829; T→C modification of nucleotide 832; T→C modification of nucleotide 835; A→G modification of nucleotide 838; A→G modification of nucleotide 841; A→C modification of nucleotide 844; T→C modification of nucleotide 847; T→C modification of nucleotide 850; T→C modification of nucleotide 853; T→C modification of nucleotide 859; G→C modification of nucleotide 862; T→C modification of nucleotide 866; T→C modification of nucleotide 871; T→G modification of nucleotide 878; T→C modification of nucleotide 883; T→C modification of nucleotide 886; T→C modification of nucleotide 889; T→C modification of nucleotide 896; A→G modification of nucleotide 898; T→C modification of nucleotide 910; T→C modification of nucleotide 916; A→C modification of nucleotide 919; T→C modification of nucleotide 925; T→C modification of nucleotide 928; A→G modification of nucleotide 940; T→G modification of nucleotide 946; T→C modification of nucleotide 949; A→C modification of nucleotide 952; A→C modification of nucleotide 955; T→C modification of nucleotide 961; T→C modification of nucleotide 964; T→C modification of nucleotide 968; T→C modification of nucleotide 973; T→C modification of nucleotide 976; T→C modification of nucleotide 982; A→C modification of nucleotide 985; T→C modification of nucleotide 988; C→G modification of nucleotide 991; T→G modification of nucleotide 1000; T→C modification of nucleotide 1001; A→G modification of nucleotide 1003; T→C modification of nucleotide 1004; A→G modification of nucleotide 1006; T→C modification of nucleotide 1009; A→G modification of nucleotide 1012; A→G modification of nucleotide 1015; A→C modification of nucleotide 1018; T→C modification of nucleotide 1021; A→G modification of nucleotide 1024; A→C modification of nucleotide 1030; T→C modification of nucleotide 1036; T→C modification of nucleotide 1037; A→G modification of nucleotide 1039; T→C modification of nucleotide 1042; T→C modification of nucleotide 1045; G→C modification of nucleotide 1048; A→G modification of nucleotide 1057; T→G modification of nucleotide 1063; T→C modification of nucleotide 1067; C→G modification of nucleotide 1072; A→G modification of nucleotide 1078; A→C modification of nucleotide 1081; T→C modification of nucleotide 1084; T→G modification of nucleotide 1087; A→G modification of nucleotide 1090; T→C modification of nucleotide 1094; T→C modification of nucleotide 1099; T→C modification of nucleotide 1102; G→C modification of nucleotide 1105; A→C modification of nucleotide 1108; T→G modification of nucleotide 1117; A→G modification of nucleotide 1120; T→C modification of nucleotide 1123; T→C modification of nucleotide 1124; A→G modification of nucleotide 1126; T→C modification of nucleotide 1129; T→C modification of nucleotide 1132; A→G modification of nucleotide 1141; A→G modification of nucleotide 1144; A→T modification of nucleotide 1145; G→C modification of nucleotide 1146; C→G modification of nucleotide 1153; A→C modification of nucleotide 1159; T→C modification of nucleotide 1165; A→T modification of nucleotide 1166; G→C modification of nucleotide 1167; T→C modification of nucleotide 1168; T→C modification of nucleotide 1171; A→C modification of nucleotide 1174; A→G modification of nucleotide 1180; T→C modification of nucleotide 1183; T→C modification of nucleotide 1184; A→G modification of nucleotide 1186; A→G modification of nucleotide 1189; T→C modification of nucleotide 1192; A→C modification of nucleotide 1195; A→C modification of nucleotide 1199; G→C modification of nucleotide 1201; T→C modification of nucleotide 1207; T→C modification of nucleotide 1210; A→G modification of nucleotide 1216; A→G modification of nucleotide 1219; A→G modification of nucleotide 1222; A→C modification of nucleotide 1234; A→T modification of nucleotide 1235; G→C modification of nucleotide 1236; A→C modification of nucleotide 1238; A→C modification of nucleotide 1240; A→G modification of nucleotide 1255; T→C modification of nucleotide 1258; C→G modification of nucleotide 1261; T→C modification of nucleotide 1267; A→T modification of nucleotide 1268; G→C modification of nucleotide 1269; T→C modification of nucleotide 1270; T→C modification of nucleotide 1273; G→C modification of nucleotide 1279; T→C modification of nucleotide 1282; T→C modification of nucleotide 1285; A→G modification of nucleotide 1288; A→G modification of nucleotide 1291; T→C modification of nucleotide 1294; T→C modification of nucleotide 1297; A→C modification of nucleotide 1300; A→G modification of nucleotide 1306; T→C modification of nucleotide 1309; T→C modification of nucleotide 1310; A→G modification of nucleotide 1312; A→C modification of nucleotide 1315; T→G modification of nucleotide 1318; A→C modification of nucleotide 1327; T→C modification of nucleotide 1330; A→C modification of nucleotide 1333; A→T modification of nucleotide 1334; G→C modification of nucleotide 1335; T→C modification of nucleotide 1336; A→G modification of nucleotide 1339; T→C modification of nucleotide 1342; T→G modification of nucleotide 1345; T→G modification of nucleotide 1348; T→C modification of nucleotide 1357; T→G modification of nucleotide 1360; T→C modification of nucleotide 1366; C→G modification of nucleotide 1369; A→C modification of nucleotide 1370; G→C modification of nucleotide 1372; T→C modification of nucleotide 1375; T→G modification of nucleotide 1378; A→C modification of nucleotide 1381; T→C modification of nucleotide 1384; T→C modification of nucleotide 1387; A→C modification of nucleotide 1390; T→C modification of nucleotide 1393; A→G modification of nucleotide 1396; A→G modification of nucleotide 1402; G→C modification of nucleotide 1405; A→G modification of nucleotide 1411; T→C modification of nucleotide 1414; C→G modification of nucleotide 1417; T→G modification of nucleotide 1420; G→C modification of nucleotide 1423; T→C modification of nucleotide 1426; T→G modification of nucleotide 1429; T→G modification of nucleotide 1432; T→G modification of nucleotide 1438; T→C modification of nucleotide 1444; A→C modification of nucleotide 1447; T→C modification of nucleotide 1450; T→C modification of nucleotide 1453; A→C modification of nucleotide 1456; T→C modification of nucleotide 1459; C→G modification of nucleotide 1465; T→C modification of nucleotide 1471; A→C modification of nucleotide 1474; T→C modification of nucleotide 1477; A→C modification of nucleotide 1480; T→C modification of nucleotide 1483; T→G modification of nucleotide 1486; T→C modification of nucleotide 1489; C→G modification of nucleotide 1495; T→C modification of nucleotide 1498; T→C modification of nucleotide 1504; T→C modification of nucleotide 1507; T→C modification of nucleotide 1510; C→G modification of nucleotide 1528; A→G modification of nucleotide 1534; T→C modification of nucleotide 1540; T→C modification of nucleotide 1543; A→C modification of nucleotide 1546; T→C modification of nucleotide 1552; T→G modification of nucleotide 1555; T→C modification of nucleotide 1561; T→C modification of nucleotide 1564; C→G modification of nucleotide 1567; T→G modification of nucleotide 1570; G→C modification of nucleotide 1573; T→C modification of nucleotide 1576; T→C modification of nucleotide 1579; A→C modification of nucleotide 1583; G→C modification of nucleotide 1585; A→G modification of nucleotide 1588; T→C modification of nucleotide 1597; T→C modification of nucleotide 1603; T→C modification of nucleotide 1612; T→C modification of nucleotide 1615; T→G modification of nucleotide 1618; T→C modification of nucleotide 1621; T→C modification of nucleotide 1624; A→T modification of nucleotide 1631; G→C modification of nucleotide 1632; A→C modification of nucleotide 1636; A→G modification of nucleotide 1642; A→G modification of nucleotide 1645; A→C modification of nucleotide 1648; T→C modification of nucleotide 1663; A→C modification of nucleotide 1666; T→G modification of nucleotide 1669; T→G modification of nucleotide 1675; T→C modification of nucleotide 1681; A→C modification of nucleotide 1684; A→G modification of nucleotide 1687; A→C modification of nucleotide 1690; T→G modification of nucleotide 1693; T→G modification of nucleotide 1699; C→G modification of nucleotide 1705; T→C modification of nucleotide 1706; A→G modification of nucleotide 1708; A→C modification of nucleotide 1714; A→C modification of nucleotide 1717; T→C modification of nucleotide 1729; A→C modification of nucleotide 1735; T→C modification of nucleotide 1741; T→C modification of nucleotide 1744; T→G modification of nucleotide 1747; A→C modification of nucleotide 1750; T→C modification of nucleotide 1751; T→C modification of nucleotide 1756; A→C modification of nucleotide 1759; T→C modification of nucleotide 1762; T→G modification of nucleotide 1765; T→G modification of nucleotide 1771; T→C modification of nucleotide 1774; A→C modification of nucleotide 1780; T→C modification of nucleotide 1783; T→C modification of nucleotide 1786; A→C modification of nucleotide 1789; T→C modification of nucleotide 1792; T→C modification of nucleotide 1793; T→C modification of nucleotide 1798; T→C modification of nucleotide 1807; T→C modification of nucleotide 1813; A→C modification of nucleotide 1816; A→C modification of nucleotide 1819; A→G modification of nucleotide 1822; T→G modification of nucleotide 1825; T→C modification of nucleotide 1829; T→G modification of nucleotide 1834; T→G modification of nucleotide 1837; T→C modification of nucleotide 1840; A→C modification of nucleotide 1843; T→C modification of nucleotide 1846; T→C modification of nucleotide 1852; T→C modification of nucleotide 1864; G→C modification of nucleotide 1867; T→C modification of nucleotide 1876; G→C modification of nucleotide 1879; A→C modification of nucleotide 1882; A→G modification of nucleotide 1891; T→C modification of nucleotide 1894; G→C modification of nucleotide 1903; C→G modification of nucleotide 1906; A→G modification of nucleotide 1915; A→G modification of nucleotide 1918; A→C modification of nucleotide 1921; T→G modification of nucleotide 1924; A→G modification of nucleotide 1927; T→C modification of nucleotide 1930; T→C modification of nucleotide 1933; A→C modification of nucleotide 1939; T→G modification of nucleotide 1945; A→G modification of nucleotide 1954; T→C modification of nucleotide 1957; G→C modification of nucleotide 1960; T→C modification of nucleotide 1963; T→G modification of nucleotide 1966; A→C modification of nucleotide 1975; T→C modification of nucleotide 1978; T→G modification of nucleotide 1981; A→C modification of nucleotide 1985; A→C modification of nucleotide 1987; A→C modification of nucleotide 1990; A→G modification of nucleotide 1993; T→C modification of nucleotide 1996; T→C modification of nucleotide 2002; A→C modification of nucleotide 2005; T→C modification of nucleotide 2006; T→C modification of nucleotide 2011; T→G modification of nucleotide 2023; T→C modification of nucleotide 2026; T→C modification of nucleotide 2029; T→C modification of nucleotide 2035; A→G modification of nucleotide 2038; A→G modification of nucleotide 2047; A→C modification of nucleotide 2048; A→C modification of nucleotide 2050; A→G modification of nucleotide 2053; T→C modification of nucleotide 2056; A→C modification of nucleotide 2059; G→C modification of nucleotide 2062; C→G modification of nucleotide 2065; C→G modification of nucleotide 2068; T→C modification of nucleotide 2071; A→G modification of nucleotide 2074; T→G modification of nucleotide 2077; T→C modification of nucleotide 2080; A→G modification of nucleotide 2092; T→C modification of nucleotide 2095; A→C modification of nucleotide 2098; A→G modification of nucleotide 2101; T→C modification of nucleotide 2104; T→C modification of nucleotide 2107; A→G modification of nucleotide 2110; T→C modification of nucleotide 2113; A→C modification of nucleotide 2119; A→C modification of nucleotide 2125; T→C modification of nucleotide 2128; T→C modification of nucleotide 2131; A→C modification of nucleotide 2137; T→C modification of nucleotide 2141; A→G modification of nucleotide 2143; T→G modification of nucleotide 2146; T→C modification of nucleotide 2152; A→C modification of nucleotide 2153; G→C modification of nucleotide 2155; T→C modification of nucleotide 2158; A→G modification of nucleotide 2161; T→C modification of nucleotide 2164; G→C modification of nucleotide 2170; A→C modification of nucleotide 2173; T→C modification of nucleotide 2176; A→C modification of nucleotide 2179; T→C modification of nucleotide 2182; T→C modification of nucleotide 2185; T→G modification of nucleotide 2188; T→C modification of nucleotide 2189; A→C modification of nucleotide 2197; T→C modification of nucleotide 2203; A→C modification of nucleotide 2215; A→G modification of nucleotide 2218; T→C modification of nucleotide 2221; A→G modification of nucleotide 2230; G→C modification of nucleotide 2233; T→G modification of nucleotide 2236; A→C modification of nucleotide 2239; T→G modification of nucleotide 2242; A→T modification of nucleotide 2252; G→C modification of nucleotide 2253; T→C modification of nucleotide 2257; C→G modification of nucleotide 2260; A→C modification of nucleotide 2264; A→C modification of nucleotide 2266; A→C modification of nucleotide 2269; A→C modification of nucleotide 2275; T→C modification of nucleotide 2281; T→C modification of nucleotide 2287; T→C modification of nucleotide 2293; T→C modification of nucleotide 2296; T→C modification of nucleotide 2299; T→G modification of nucleotide 2302; A→C modification of nucleotide 2305; T→G modification of nucleotide 2308; T→C modification of nucleotide 2311; T→C modification of nucleotide 2314; T→C modification of nucleotide 2317; A→C modification of nucleotide 2326; A→G modification of nucleotide 2329; T→G modification of nucleotide 2335; A→C modification of nucleotide 2338; T→C modification of nucleotide 2341; T→C modification of nucleotide 2350; T→C modification of nucleotide 2353; T→C modification of nucleotide 2356; A→T modification of nucleotide 2357; G→C modification of nucleotide 2358; T→C modification of nucleotide 2362; T→C modification of nucleotide 2365; T→C modification of nucleotide 2368; T→C modification of nucleotide 2371; T→C modification of nucleotide 2377; T→C modification of nucleotide 2380; A→C modification of nucleotide 2383; T→C modification of nucleotide 2386; T→C modification of nucleotide 2395; T→C modification of nucleotide 2399; T→C modification of nucleotide 2404; A→G modification of nucleotide 2422; T→C modification of nucleotide 2425; T→C modification of nucleotide 2434; T→C modification of nucleotide 2447; T→C modification of nucleotide 2449; T→C modification of nucleotide 2452; T→C modification of nucleotide 2458; A→C modification of nucleotide 2461; T→G modification of nucleotide 2464; T→C modification of nucleotide 2467; G→C modification of nucleotide 2470; A→C modification of nucleotide 2476; T→C modification of nucleotide 2479; G→C modification of nucleotide 2485; T→C modification of nucleotide 2488; C→G modification of nucleotide 2500; T→C modification of nucleotide 2506; G→C modification of nucleotide 2509; T→C modification of nucleotide 2515; A→C modification of nucleotide 2518; T→C modification of nucleotide 2521; A→T modification of nucleotide 2525; G→C modification of nucleotide 2526; T→C modification of nucleotide 2530; T→C modification of nucleotide 2545; T→C modification of nucleotide 2548; T→C modification of nucleotide 2551; and deletion of nucleotides 2558-3197;

›DETAILED DESCRIPTION · 27 of 32

k) a sequence encoding yes-associated protein 1 (YAP) and comprising one or more of the following modifications relative to SEQ ID NO: 22:

deletion of nucleotides 1-402; T→C modification of nucleotide 408; G→C modification of nucleotide 414; G→C modification of nucleotide 423; G→C modification of nucleotide 426; T→C modification of nucleotide 429; A→G modification of nucleotide 432; G→C modification of nucleotide 435; A→G modification of nucleotide 450; G→C modification of nucleotide 453; G→C modification of nucleotide 459; T→C modification of nucleotide 462; G→C modification of nucleotide 465; G→C modification of nucleotide 474; G→C modification of nucleotide 480; G→C modification of nucleotide 489; G→C modification of nucleotide 492; A→C modification of nucleotide 498; G→C modification of nucleotide 504; A→G modification of nucleotide 507; G→C modification of nucleotide 510; A→C modification of nucleotide 513; G→C modification of nucleotide 519; G→C modification of nucleotide 522; G→C modification of nucleotide 531; G→C modification of nucleotide 534; G→C modification of nucleotide 537; A→C modification of nucleotide 543; G→C modification of nucleotide 555; T→C modification of nucleotide 558; C→G modification of nucleotide 573; G→C modification of nucleotide 579; G→C modification of nucleotide 585; G→C modification of nucleotide 603; C→G modification of nucleotide 606; C→G modification of nucleotide 618; G→C modification of nucleotide 633; A→C modification of nucleotide 661; G→C modification of nucleotide 663; C→G modification of nucleotide 666; G→C modification of nucleotide 669; G→C modification of nucleotide 696; G→C modification of nucleotide 699; A→G modification of nucleotide 708; A→C modification of nucleotide 720; A→T modification of nucleotide 727; G→C modification of nucleotide 728; T→C modification of nucleotide 729; T→C modification of nucleotide 732; T→C modification of nucleotide 735; A→C modification of nucleotide 738; T→C modification of nucleotide 744; A→C modification of nucleotide 747; A→C modification of nucleotide 750; T→C modification of nucleotide 759; A→C modification of nucleotide 762; T→C modification of nucleotide 768; T→G modification of nucleotide 771; A→C modification of nucleotide 774; T→C modification of nucleotide 777; T→C modification of nucleotide 780; T→G modification of nucleotide 781; T→C modification of nucleotide 786; A→C modification of nucleotide 789; T→C modification of nucleotide 792; T→C modification of nucleotide 795; T→C modification of nucleotide 802; A→C modification of nucleotide 807; T→C modification of nucleotide 810; T→G modification of nucleotide 813; T→C modification of nucleotide 816; T→C modification of nucleotide 819; T→C modification of nucleotide 822; A→C modification of nucleotide 825; T→C modification of nucleotide 837; A→C modification of nucleotide 840; A→G modification of nucleotide 843; C→G modification of nucleotide 846; T→C modification of nucleotide 849; A→C modification of nucleotide 855; A→C modification of nucleotide 858; T→C modification of nucleotide 861; A→C modification of nucleotide 864; A→C modification of nucleotide 870; T→C modification of nucleotide 873; T→C modification of nucleotide 879; T→G modification of nucleotide 882; A→C modification of nucleotide 885; T→C modification of nucleotide 891; T→C modification of nucleotide 894; T→C modification of nucleotide 897; A→C modification of nucleotide 903; T→C modification of nucleotide 906; T→C modification of nucleotide 909; T→C modification of nucleotide 912; A→G modification of nucleotide 915; T→C modification of nucleotide 918; A→C modification of nucleotide 924; A→C modification of nucleotide 927; T→C modification of nucleotide 930; A→C modification of nucleotide 942; A→C modification of nucleotide 948; T→C modification of nucleotide 951; T→C modification of nucleotide 954; T→C modification of nucleotide 957; A→C modification of nucleotide 961; A→C modification of nucleotide 963; T→C modification of nucleotide 970; A→G modification of nucleotide 972; T→C modification of nucleotide 975; T→C modification of nucleotide 984; A→C modification of nucleotide 990; A→C modification of nucleotide 993; A→C modification of nucleotide 996; A→C modification of nucleotide 1009; G→C modification of nucleotide 1011; C→G modification of nucleotide 1038; A→C modification of nucleotide 1041; A→T modification of nucleotide 1051; G→C modification of nucleotide 1052; T→C modification of nucleotide 1053; A→C modification of nucleotide 1056; A→C modification of nucleotide 1059; T→C modification of nucleotide 1071; G→C modification of nucleotide 1083; T→C modification of nucleotide 1086; A→C modification of nucleotide 1089; T→C modification of nucleotide 1092; T→C modification of nucleotide 1095; T→G modification of nucleotide 1098; T→C modification of nucleotide 1101; T→C modification of nucleotide 1104; A→C modification of nucleotide 1107; A→G modification of nucleotide 1113; A→G modification of nucleotide 1116; T→C modification of nucleotide 1125; T→C modification of nucleotide 1131; A→C modification of nucleotide 1134; A→G modification of nucleotide 1137; T→C modification of nucleotide 1140; T→C modification of nucleotide 1146; A→C modification of nucleotide 1149; T→C modification of nucleotide 1155; T→C modification of nucleotide 1173; A→G modification of nucleotide 1179; A→C modification of nucleotide 1185; A→C modification of nucleotide 1186; G→C modification of nucleotide 1188; T→G modification of nucleotide 1191; T→C modification of nucleotide 1197; T→C modification of nucleotide 1200; T→C modification of nucleotide 1203; A→C modification of nucleotide 1216; A→C modification of nucleotide 1218; A→T modification of nucleotide 1222; G→C modification of nucleotide 1223; T→C modification of nucleotide 1224; A→T modification of nucleotide 1228; G→C modification of nucleotide 1229; T→C modification of nucleotide 1230; T→C modification of nucleotide 1233; A→C modification of nucleotide 1236; A→G modification of nucleotide 1242; A→C modification of nucleotide 1248; A→C modification of nucleotide 1251; T→C modification of nucleotide 1260; A→T modification of nucleotide 1267; G→C modification of nucleotide 1268; A→C modification of nucleotide 1272; A→C modification of nucleotide 1278; C→G modification of nucleotide 1284; T→C modification of nucleotide 1290; A→T modification of nucleotide 1294; G→C modification of nucleotide 1295; A→G modification of nucleotide 1314; A→C modification of nucleotide 1323; A→G modification of nucleotide 1332; A→C modification of nucleotide 1351; G→C modification of nucleotide 1353; G→C modification of nucleotide 1359; A→G modification of nucleotide 1365; A→G modification of nucleotide 1371; A→G modification of nucleotide 1374; T→G modification of nucleotide 1380; G→C modification of nucleotide 1383; T→C modification of nucleotide 1390; A→G modification of nucleotide 1392; T→C modification of nucleotide 1401; A→T modification of nucleotide 1402; G→C modification of nucleotide 1403; T→C modification of nucleotide 1408; A→G modification of nucleotide 1410; A→C modification of nucleotide 1413; A→C modification of nucleotide 1416; T→C modification of nucleotide 1428; T→C modification of nucleotide 1431; G→C modification of nucleotide 1434; T→C modification of nucleotide 1437; A→G modification of nucleotide 1440; T→C modification of nucleotide 1443; A→C modification of nucleotide 1446; T→C modification of nucleotide 1452; T→C modification of nucleotide 1455; G→C modification of nucleotide 1461; T→C modification of nucleotide 1467; A→G modification of nucleotide 1473; T→C modification of nucleotide 1474; A→C modification of nucleotide 1477; A→C modification of nucleotide 1479; A→C modification of nucleotide 1482; G→C modification of nucleotide 1488; T→C modification of nucleotide 1494; A→T modification of nucleotide 1495; G→C modification of nucleotide 1496; A→C modification of nucleotide 1500; T→C modification of nucleotide 1503; T→C modification of nucleotide 1506; T→G modification of nucleotide 1512; A→T modification of nucleotide 1516; G→C modification of nucleotide 1517; T→C modification of nucleotide 1518; T→C modification of nucleotide 1527; T→C modification of nucleotide 1533; A→C modification of nucleotide 1536; T→C modification of nucleotide 1539; A→T modification of nucleotide 1543; G→C modification of nucleotide 1544; T→C modification of nucleotide 1545; A→C modification of nucleotide 1548; A→T modification of nucleotide 1552; G→C modification of nucleotide 1553; T→C modification of nucleotide 1554; A→C modification of nucleotide 1557; A→G modification of nucleotide 1560; A→T modification of nucleotide 1561; G→C modification of nucleotide 1562; A→T modification of nucleotide 1567; G→C modification of nucleotide 1568; A→T modification of nucleotide 1570; G→C modification of nucleotide 1571; A→T modification of nucleotide 1576; G→C modification of nucleotide 1577; T→C modification of nucleotide 1578; C→G modification of nucleotide 1581; T→C modification of nucleotide 1584; A→C modification of nucleotide 1587; A→C modification of nucleotide 1593; T→C modification of nucleotide 1596; A→T modification of nucleotide 1609; G→C modification of nucleotide 1610; T→C modification of nucleotide 1611; T→C modification of nucleotide 1617; T→C modification of nucleotide 1626; A→C modification of nucleotide 1629; T→C modification of nucleotide 1632; T→C modification of nucleotide 1635; T→C modification of nucleotide 1638; A→G modification of nucleotide 1647; A→T modification of nucleotide 1648; G→C modification of nucleotide 1649; A→C modification of nucleotide 1662; T→C modification of nucleotide 1674; A→C modification of nucleotide 1680; T→G modification of nucleotide 1689; A→G modification of nucleotide 1692; T→C modification of nucleotide 1698; T→C modification of nucleotide 1701; G→C modification of nucleotide 1704; A→C modification of nucleotide 1707; T→C modification of nucleotide 1710; T→G modification of nucleotide 1719; A→C modification of nucleotide 1722; A→C modification of nucleotide 1725; A→G modification of nucleotide 1731; A→C modification of nucleotide 1734; T→C modification of nucleotide 1737; A→C modification of nucleotide 1740; A→C modification of nucleotide 1749; A→G modification of nucleotide 1752; A→C modification of nucleotide 1755; A→C modification of nucleotide 1770; A→T modification of nucleotide 1771; G→C modification of nucleotide 1772; T→C modification of nucleotide 1773; A→G modification of nucleotide 1782; T→C modification of nucleotide 1785; T→C modification of nucleotide 1786; A→T modification of nucleotide 1789; G→C modification of nucleotide 1790; T→C modification of nucleotide 1791; T→C modification of nucleotide 1794; T→G modification of nucleotide 1803; T→C modification of nucleotide 1806; T→C modification of nucleotide 1818; T→G modification of nucleotide 1821; T→C modification of nucleotide 1822; T→C modification of nucleotide 1827; A→G modification of nucleotide 1839; T→C modification of nucleotide 1842; A→G modification of nucleotide 1845; A→G modification of nucleotide 1848; A→T modification of nucleotide 1849; G→C modification of nucleotide 1850; T→C modification of nucleotide 1854; T→G modification of nucleotide 1857; A→C modification of nucleotide 1860; and deletion of nucleotides 1864-5353;

›DETAILED DESCRIPTION · 28 of 32

l) a sequence encoding wingless-type MMTV integration site family, member 2 (WNT2) and comprising one or more of the following modifications relative to SEQ ID NO: 23:

deletion of nucleotides 1-158; C→G modification of nucleotide 167; T→C modification of nucleotide 170; C→G modification of nucleotide 173; T→C modification of nucleotide 176; A→C modification of nucleotide 179; C→G modification of nucleotide 188; C→G modification of nucleotide 194; T→C modification of nucleotide 197; C→G modification of nucleotide 203; T→C modification of nucleotide 204; C→G modification of nucleotide 215; T→C modification of nucleotide 221; C→G modification of nucleotide 227; A→T modification of nucleotide 228; G→C modification of nucleotide 229; T→C modification of nucleotide 233; A→C modification of nucleotide 236; A→C modification of nucleotide 249; A→C modification of nucleotide 251; T→C modification of nucleotide 254; A→C modification of nucleotide 257; T→C modification of nucleotide 260; A→C modification of nucleotide 270; G→C modification of nucleotide 272; T→C modification of nucleotide 281; T→C modification of nucleotide 287; A→C modification of nucleotide 293; A→T modification of nucleotide 303; G→C modification of nucleotide 304; G→C modification of nucleotide 308; T→C modification of nucleotide 314; A→C modification of nucleotide 329; A→C modification of nucleotide 335; T→C modification of nucleotide 338; T→C modification of nucleotide 347; T→C modification of nucleotide 353; T→C modification of nucleotide 362; T→C modification of nucleotide 368; T→C modification of nucleotide 377; A→C modification of nucleotide 380; A→G modification of nucleotide 389; T→C modification of nucleotide 407; A→C modification of nucleotide 432; A→C modification of nucleotide 434; T→C modification of nucleotide 437; A→T modification of nucleotide 441; G→C modification of nucleotide 442; C→G modification of nucleotide 446; T→C modification of nucleotide 449; G→C modification of nucleotide 455; T→C modification of nucleotide 458; C→G modification of nucleotide 461; C→G modification of nucleotide 464; A→C modification of nucleotide 467; A→T modification of nucleotide 468; G→C modification of nucleotide 469; T→C modification of nucleotide 470; A→T modification of nucleotide 471; G→C modification of nucleotide 472; T→C modification of nucleotide 473; G→C modification of nucleotide 476; A→G modification of nucleotide 479; G→C modification of nucleotide 482; T→C modification of nucleotide 488; T→G modification of nucleotide 491; T→C modification of nucleotide 503; A→C modification of nucleotide 506; T→C modification of nucleotide 509; T→G modification of nucleotide 515; A→G modification of nucleotide 518; T→C modification of nucleotide 521; A→C modification of nucleotide 531; G→C modification of nucleotide 533; T→C modification of nucleotide 539; A→T modification of nucleotide 540; G→C modification of nucleotide 541; A→G modification of nucleotide 545; A→C modification of nucleotide 548; A→G modification of nucleotide 551; T→C modification of nucleotide 552; A→G modification of nucleotide 554; T→C modification of nucleotide 569; T→C modification of nucleotide 572; A→C modification of nucleotide 575; A→G modification of nucleotide 584; A→C modification of nucleotide 587; A→T modification of nucleotide 588; G→C modification of nucleotide 589; T→C modification of nucleotide 590; A→T modification of nucleotide 600; G→C modification of nucleotide 601; A→G modification of nucleotide 605; T→C modification of nucleotide 623; A→T modification of nucleotide 630; G→C modification of nucleotide 631; T→C modification of nucleotide 632; T→C modification of nucleotide 638; T→C modification of nucleotide 641; G→C modification of nucleotide 650; T→C modification of nucleotide 659; T→C modification of nucleotide 665; T→C modification of nucleotide 671; A→G modification of nucleotide 674; T→C modification of nucleotide 677; A→C modification of nucleotide 687; G→C modification of nucleotide 689; A→G modification of nucleotide 692; T→C modification of nucleotide 701; A→C modification of nucleotide 705; A→C modification of nucleotide 707; T→G modification of nucleotide 722; A→C modification of nucleotide 732; A→C modification of nucleotide 734; T→C modification of nucleotide 737; A→C modification of nucleotide 740; A→C modification of nucleotide 741; G→C modification of nucleotide 743; T→C modification of nucleotide 749; A→G modification of nucleotide 752; T→C modification of nucleotide 762; A→G modification of nucleotide 767; A→G modification of nucleotide 770; A→G modification of nucleotide 773; T→C modification of nucleotide 782; T→C modification of nucleotide 785; T→C modification of nucleotide 788; A→T modification of nucleotide 792; G→C modification of nucleotide 793; T→C modification of nucleotide 794; T→C modification of nucleotide 803; T→C modification of nucleotide 806; A→C modification of nucleotide 810; G→C modification of nucleotide 812; A→C modification of nucleotide 815; T→C modification of nucleotide 833; A→C modification of nucleotide 840; G→C modification of nucleotide 842; A→G modification of nucleotide 845; A→C modification of nucleotide 848; T→C modification of nucleotide 857; C→G modification of nucleotide 860; A→C modification of nucleotide 864; G→C modification of nucleotide 866; T→C modification of nucleotide 875; G→C modification of nucleotide 878; A→G modification of nucleotide 890; C→G modification of nucleotide 893; T→C modification of nucleotide 905; T→C modification of nucleotide 911; T→C modification of nucleotide 920; A→G modification of nucleotide 923; T→C modification of nucleotide 929; A→C modification of nucleotide 933; G→C modification of nucleotide 935; T→C modification of nucleotide 938; A→C modification of nucleotide 947; G→C modification of nucleotide 950; A→G modification of nucleotide 953; T→C modification of nucleotide 956; C→G modification of nucleotide 962; T→C modification of nucleotide 968; T→C modification of nucleotide 971; T→C modification of nucleotide 977; T→C modification of nucleotide 980; A→C modification of nucleotide 983; T→C modification of nucleotide 992; A→C modification of nucleotide 996; G→C modification of nucleotide 998; A→C modification of nucleotide 1004; A→C modification of nucleotide 1010; T→C modification of nucleotide 1022; A→C modification of nucleotide 1025; G→C modification of nucleotide 1028; T→C modification of nucleotide 1034; T→C modification of nucleotide 1044; T→C modification of nucleotide 1049; A→C modification of nucleotide 1055; A→T modification of nucleotide 1065; G→C modification of nucleotide 1066; A→G modification of nucleotide 1073; T→G modification of nucleotide 1076; T→C modification of nucleotide 1082; T→C modification of nucleotide 1085; G→C modification of nucleotide 1088; A→C modification of nucleotide 1089; A→C modification of nucleotide 1091; T→C modification of nucleotide 1097; A→C modification of nucleotide 1103; C→G modification of nucleotide 1112; G→C modification of nucleotide 1118; T→C modification of nucleotide 1130; T→C modification of nucleotide 1136; A→G modification of nucleotide 1139; T→C modification of nucleotide 1154; T→C modification of nucleotide 1166; A→C modification of nucleotide 1199; A→T modification of nucleotide 1215; G→C modification of nucleotide 1216; T→C modification of nucleotide 1217; and deletion of nucleotides 1229-2115; and/or

›DETAILED DESCRIPTION · 29 of 32

m) a sequence encoding wingless-type MMTV integration site family, member 9B (WNT9b) and comprising one or more of the following modifications relative to SEQ ID NO: 24:

deletion of nucleotides 1-52; G→C modification of nucleotide 64; G→C modification of nucleotide 70; T→C modification of nucleotide 82; G→C modification of nucleotide 85; G→C modification of nucleotide 88; T→C modification of nucleotide 106; T→C modification of nucleotide 112; T→C modification of nucleotide 145; T→C modification of nucleotide 148; C→G modification of nucleotide 154; A→C modification of nucleotide 160; A→C modification of nucleotide 169; T→C modification of nucleotide 178; G→C modification of nucleotide 181; A→C modification of nucleotide 184; A→C modification of nucleotide 187; G→C modification of nucleotide 193; A→C modification of nucleotide 196; T→C modification of nucleotide 202; T→C modification of nucleotide 205; T→C modification of nucleotide 208; T→C modification of nucleotide 223; A→G modification of nucleotide 229; A→C modification of nucleotide 242; G→C modification of nucleotide 244; G→C modification of nucleotide 247; C→G modification of nucleotide 259; A→C modification of nucleotide 263; G→C modification of nucleotide 265; G→C modification of nucleotide 268; T→C modification of nucleotide 283; A→C modification of nucleotide 293; G→C modification of nucleotide 295; T→C modification of nucleotide 298; T→C modification of nucleotide 301; A→C modification of nucleotide 304; G→C modification of nucleotide 313; A→G modification of nucleotide 322; T→C modification of nucleotide 325; A→C modification of nucleotide 338; G→C modification of nucleotide 340; A→T modification of nucleotide 359; G→C modification of nucleotide 360; G→C modification of nucleotide 370; A→C modification of nucleotide 371; G→C modification of nucleotide 373; T→C modification of nucleotide 376; C→G modification of nucleotide 385; A→C modification of nucleotide 389; A→C modification of nucleotide 391; T→C modification of nucleotide 397; G→C modification of nucleotide 406; T→C modification of nucleotide 418; A→C modification of nucleotide 421; T→C modification of nucleotide 427; A→C modification of nucleotide 430; T→C modification of nucleotide 433; C→G modification of nucleotide 439; G→C modification of nucleotide 442; T→C modification of nucleotide 445; A→C modification of nucleotide 448; T→C modification of nucleotide 455; G→C modification of nucleotide 457; A→T modification of nucleotide 464; G→C modification of nucleotide 465; T→C modification of nucleotide 466; T→C modification of nucleotide 469; G→C modification of nucleotide 472; T→C modification of nucleotide 487; T→C modification of nucleotide 490; T→C modification of nucleotide 493; A→C modification of nucleotide 505; A→T modification of nucleotide 515; G→C modification of nucleotide 516; G→C modification of nucleotide 520; T→C modification of nucleotide 538; T→C modification of nucleotide 544; T→C modification of nucleotide 547; T→C modification of nucleotide 553; A→T modification of nucleotide 563; G→C modification of nucleotide 564; C→G modification of nucleotide 577; A→T modification of nucleotide 578; G→C modification of nucleotide 579; G→C modification of nucleotide 592; A→C modification of nucleotide 599; A→C modification of nucleotide 601; A→C modification of nucleotide 604; A→T modification of nucleotide 605; G→C modification of nucleotide 606; A→C modification of nucleotide 617; G→C modification of nucleotide 619; G→C modification of nucleotide 622; A→C modification of nucleotide 623; G→C modification of nucleotide 625; T→C modification of nucleotide 628; T→C modification of nucleotide 661; A→T modification of nucleotide 668; G→C modification of nucleotide 669; A→C modification of nucleotide 677; A→C modification of nucleotide 679; A→C modification of nucleotide 682; T→C modification of nucleotide 697; T→C modification of nucleotide 700; A→C modification of nucleotide 706; T→C modification of nucleotide 715; T→C modification of nucleotide 718; T→G modification of nucleotide 721; T→C modification of nucleotide 724; T→C modification of nucleotide 730; C→G modification of nucleotide 742; G→C modification of nucleotide 748; T→C modification of nucleotide 751; A→G modification of nucleotide 778; T→C modification of nucleotide 784; G→C modification of nucleotide 790; T→C modification of nucleotide 793; C→G modification of nucleotide 796; A→T modification of nucleotide 806; G→C modification of nucleotide 807; T→C modification of nucleotide 808; T→C modification of nucleotide 824; T→C modification of nucleotide 829; T→C modification of nucleotide 832; A→G modification of nucleotide 841; T→C modification of nucleotide 853; A→C modification of nucleotide 859; T→C modification of nucleotide 862; T→C modification of nucleotide 865; A→G modification of nucleotide 880; T→C modification of nucleotide 886; T→C modification of nucleotide 889; G→C modification of nucleotide 895; C→G modification of nucleotide 904; A→G modification of nucleotide 913; T→C modification of nucleotide 916; T→C modification of nucleotide 919; A→T modification of nucleotide 923; G→C modification of nucleotide 924; G→C modification of nucleotide 934; A→T modification of nucleotide 938; G→C modification of nucleotide 939; T→C modification of nucleotide 949; G→C modification of nucleotide 952; G→C modification of nucleotide 958; A→C modification of nucleotide 961; A→C modification of nucleotide 965; G→C modification of nucleotide 967; T→C modification of nucleotide 973; T→C modification of nucleotide 976; A→C modification of nucleotide 979; A→T modification of nucleotide 986; G→C modification of nucleotide 987; T→C modification of nucleotide 988; A→T modification of nucleotide 992; G→C modification of nucleotide 993; A→T modification of nucleotide 995; G→C modification of nucleotide 996; A→G modification of nucleotide 1000; T→C modification of nucleotide 1006; G→C modification of nucleotide 1009; A→C modification of nucleotide 1012; A→T modification of nucleotide 1028; G→C modification of nucleotide 1029; T→G modification of nucleotide 1042; T→C modification of nucleotide 1057; T→C modification of nucleotide 1096; A→C modification of nucleotide 1099; C→G modification of nucleotide 1111; T→C modification of nucleotide 1117; and deletion of nucleotides 1130-4519.

›DETAILED DESCRIPTION · 30 of 32

2. The composition of paragraph 1, wherein the at least one engineered liver regenerative factor mRNA comprises one or more of:

3. The composition of any one of the preceding paragraphs, wherein the at least one engineered liver regenerative factor mRNA further comprises at least one modified nucleoside. 4. A composition comprising at least one liver regenerative factor mRNAs comprising at least one modified nucleoside;

wherein the at least one liver regenerative factor is selected from the group consisting of:

vascular endothelial growth factor A (VEGFA); hepatocyte growth factor (HGF); growth hormone (GH); insulin-like growth factor 1 (IGF-1), epidermal growth factor (EGF); signal transducer and activator of transcription 5B (STAT5b); cyclin-dependent kinase inhibitor 1A (p21); beta catenin (CTNNB1); yes-associated protein (YAP); wingless-type MMTV integration site family, member 2 (WNT2); and wingless-type MMTV integration site family, member 9B (WNT9b).

5. The composition of any one of the preceding paragraphs, wherein the composition further comprises a carrier complexed with the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside. 6. The composition of paragraph 5, wherein the carrier is a nanoparticle. 7. The composition of paragraph 6, wherein the carrier is a polymer nanoparticle. 8. The composition of paragraph 6, wherein the nanoparticle is a lipid nanoparticle (LNP). 9. The composition of any one of the preceding paragraphs, wherein the at least one liver regenerative factor is selected from the group consisting of: VEGFA; HGF; GH; and EGF. 10. The composition of any one of the preceding paragraphs, wherein the at least one liver regenerative factor is selected from the group consisting of: HGF; GH; EGF; and p21. 11. The composition of any one of the preceding paragraphs, wherein the at least one liver regenerative factor is selected from the group consisting of: HGF; GH; and EGF. 12. The composition of any one of the preceding paragraphs, wherein the at least one liver regenerative factor comprises two or more liver regenerative factors selected from the group consisting of:

VEGFA; HGF; GH; IGF-1; EGF; STAT5bCA; p21; CNNTB1; YAP; WNT2; and WNT9b.

13. The composition of any one of the preceding paragraphs, wherein the at least one liver regenerative factor is a human liver regenerative factor or a murine liver regenerative factor. 14. The composition of any one of the preceding paragraphs, wherein the at least one modified nucleoside comprises at least one non-natural nucleoside. 15. The composition of any one of the preceding paragraphs, wherein the at least one modified nucleoside is selected from the group consisting of:

pseudouridine, N1-methylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine, pseudouridine, and mixtures thereof, 5-methyluridine, 5-methoxyuridine, and 2′-O-methyluridine.

16. The composition of any one of the preceding paragraphs, wherein the at least one modified nucleoside comprises at least one-methylpseudouridine (m1Ψ)-5′ triphosphate (TriLink). 17. The composition of any one of the preceding paragraphs, wherein the LNP comprises at least one ionizable lipid, at least one phospholipid, at least one structured lipid, and at least one polyethylene glycol (PEG)-lipid. 18. The composition of paragraph 17, wherein the at least one ionizable lipid is selected from the group consisting of:

2,2-dioleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA); dioleyl-methyl 4-dimethylaminobutyrate (DLin-MC3-DMA); and di ((Z)-non-2-en-1-yl) 9-((4-(dimethylamino) butyryl) oxy) heptadecanedioate (L319).

19. The composition of any one of paragraphs 17-18, wherein the at least one ionizable lipid has a pKA in the range of 6.0-6.5. 20. The composition of any one of paragraphs 17-19, wherein the at least one phospholipid comprises phosphatidylcholine. 21. The composition of any one of paragraphs 17-20, wherein the at least one structured lipid comprises cholesterol. 22. The composition of any one of the preceding paragraphs, wherein the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is encapsulated in the nanoparticle. 23. The composition of any one of the preceding paragraphs, wherein the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is in an aqueous solution and in admixture with an ethanolic lipid mixture at acidic pH. 24. The composition of any one of the preceding paragraphs, wherein the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside is complexed to the LNP in a way selected from the group consisting of:

encapsulation in the interior of the LNP; interspersed within the lipid bilayer of the LNP; and attached to the LNP via a linking molecule.

25. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises GH; and

the at least one engineered liver regenerative factor mRNA or the at least one liver regenerative factor mRNA comprising at least one modified nucleoside comprises a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 8.

›DETAILED DESCRIPTION · 31 of 32

26. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises EGF; and

27. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises HGF; and

28. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises p21; and

29. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises VEGF; and

30. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises IGF-1; and

31. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises IGF-1 IL-2 SP; and

32. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises secreted EGF; and

33. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises stat5b; and

34. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises beta catenin; and

35. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises YAP; and

36. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises WNT2; and

37. The composition of any one of the preceding paragraphs, wherein the liver regenerative factor comprises WNT9b; and

38. The composition of any one of the preceding paragraphs, further comprising N-acetyl cysteine (NAC). 39. A combination of the composition of any one of paragraphs 1-37 and N-acetyl cysteine (NAC). 40. A method of treating liver injury or liver disease in a subject in need thereof, the method comprising administering the composition or combination of any one of the preceding paragraphs to the subject. 41. A method of accelerating intrinsic liver repair in a subject in need thereof, the method comprising administering the composition or combination of any one of the preceding paragraphs. 42. The method of any one of the preceding paragraphs, wherein the subject is a subject in need of treatment for acute liver disease, chronic liver disease, or acetaminophen (acetyl-para-aminophenol, APAP) overdose. 43. The method of paragraph 42, wherein acute or chronic liver disease is selected from the group consisting of:

haemophilia; familial hypercholesterolemia; ornithine transcarbamylase deficiency; α-antitrypsin deficiency; non-alcoholic fatty liver disease (NAFLD); alcoholic fatty liver disease; alcohol-related liver disease (ARLD); phenylketonuria; glycogen storage disease; α1-antitrypsin deficiency; hereditary hemochromatosis; tyrosinemia type 1; argininosuccinic aciduria; hepatitis virus infection; non-viral hepatitis; autoimmune hepatitis; primary biliary cholangitis; cirrhosis, biliary atresia; liver cancer; genetic cholestasis; hemochromatosis; Gilbert syndrome; primary sclerosing cholangitis (PSC); and Wilson's disease.

›DETAILED DESCRIPTION · 32 of 32

44. The method of any one of the preceding paragraphs, further comprising administering N-acetyl cysteine (NAC) to the subject. 45. A method of engrafting cells in a liver tissue, the method comprising introducing the cells into the liver tissue and contacting the cells or the liver tissue with the composition or combination of any of one of the preceding paragraphs. 46. The method of paragraph 45, wherein the cells are primary human hepatocytes (PHH) or induced pluripotent stem cell-derived hepatocyte-like-cells (iPSC-HLCs). 47. The method of any one of paragraphs 45-46, wherein the liver tissue is a liver in a subject, the introducing comprises transplanting the cells into the liver, and the contacting comprises administering. 48. The method of paragraph 47, wherein the subject is a subject in need of treatment for acute liver disease, chronic liver disease, or genetic liver disease. 49. The method of paragraph 48, wherein acute or chronic liver disease is selected from the group consisting of:

50. The method of paragraph 48, wherein the subject is a subject in need of treatment for alpha-1 antitrypsin deficiency associated liver disease (AATD). 51. The method of any one of the preceding paragraphs, wherein the composition or combination is administered once. 52. The method of any one of the preceding paragraphs, wherein the composition or combination is administered twice or more. 53. The method of any one of the preceding paragraphs, wherein the composition or combination comprises two or more liver regenerative factor mRNAs. 54. The method of any one of the preceding paragraphs, comprising administering a first composition or combination of any one of paragraphs 1-39 comprising a first liver regenerative factor mRNA and concurrently administering a second composition or combination of any one of paragraphs 1-39 comprising a second liver regenerative factor mRNA. 55. The method of any one of the preceding paragraphs, comprising administering a first composition or combination of any one of paragraphs 1-39 comprising a first liver regenerative factor mRNA and separately administering a second composition or combination of any one of paragraphs 1-39 comprising a second liver regenerative factor mRNA. 56. The method of any one of the preceding paragraphs, wherein the administering is intravenous administration. 57. The method of any one of the preceding paragraphs, wherein the administering is via the common bile duct or to the gallbladder.

The technology described herein is further illustrated by the following examples which in no way should be construed as being further limiting.

›EXAMPLES · 1 of 2

Example 1: VEGFA mRNA-LNP Promotes Biliary Epithelial Cell-to-Hepatocyte Conversion in Acute and Chronic Liver Diseases and Reverses Steatosis and Fibrosis

The liver is known for its remarkable regenerative ability through proliferation of hepatocytes. Yet, during chronic injury or severe hepatocyte death, proliferation of hepatocytes is exhausted. To overcome this hurdle, the inventors use vascular-endothelial-growth-factor A (VEGFA) as a therapeutic means to accelerate biliary epithelial cell (BEC)-to-hepatocyte conversion. Investigation in zebrafish establishes that blocking VEGF receptors abrogates BEC-driven liver repair, while VEGFA overexpression promotes it. Delivery of VEGFA via non-integrative and safe nucleoside-modified mRNA encapsulated into lipid-nanoparticles (mRNA-LNP) in acutely or chronically injured mouse livers induces robust BEC-to-hepatocyte conversion and reversion of steatosis and fibrosis. In human and murine diseased livers, the inventors further identified VEGFA-receptor KDR-expressing BECs associated with KDR-expressing cell-derived hepatocytes. This defines KDR-expressing cells, as being BECs, as facultative progenitors. This study reveals novel therapeutic benefits of VEGFA delivered via nucleoside-modified mRNA-LNP, whose safety is widely validated with COVID-19 vaccines, for harnessing BEC-driven repair to potentially treat liver diseases.

Mortality related to end stage liver disease (ESLD) is ranked as the 12 th most common cause of death in the US. Liver transplantation remains the only treatment for ESLD, but this procedure is critically challenged by the shortage of organ donors. The remarkable ability of the liver to regenerate by proliferation of mature hepatocytes is well documented 1 , yet in the case of severe acute hepatocyte death or chronic ESLD, proliferation of mature cells becomes exhausted 2,3 due to the escalating progression of steatosis, inflammation, fibrosis and irreversible cirrhosis. In these cases, the presence of alternative precursors of hepatocytes that derive from biliary epithelial cells (BECs) has been postulated, and these cells have been referred to by various names, including liver progenitor cells (LPCs) 4 . Expansion of LPCs from the BEC compartment, a process described as ductular reaction (DR), is present in virtually all chronic and acute human liver diseases, suggesting an alternative regeneration process by which BECs proliferate and differentiate into hepatocytes 5-15 , the BEC-driven liver regeneration. The evidence for the contribution of BECs to de novo hepatocytes in humans is illustrated by the presence of hepatocytes expressing the BEC marker EpCAM emerging from highly proliferative DR areas in advanced cirrhotic livers 16 , detection of “bi-phenotypic cells” 17 or “ductular hepatocytes” 18 positive for both the BEC marker KRT19 and the hepatocyte marker HNF4α 17 or HepPar1 18 , and observation of budding of hepatocyte-like cells expressing the hepatocytic marker glutamine synthetase (GS) from BECs within the DRs 19-23 Specifically, one study quantified the emerging immature hepatocytes budding from KRT19+ BECs within the septa developed in human cirrhotic livers, and estimated that they represented up to 70% of hepatocytes 19 . Importantly, a recent study showed that aberrant GS positivity adjacent to portal tracts is significantly associated with regressed cirrhosis in humans 24 , suggesting the clinical benefit of LPC-derived hepatocytes in resolving human cirrhosis. In an attempt to lineage trace the LPCs among the DRs in human cirrhotic livers, recent studies have used mutational analysis in mitochondrial DNA encoding cytochrome c oxidase enzyme, and showed the descent of hepatocytes within monoclonal regenerative nodules from adjacent LPC-associated DRs 25 . In mice, lineage tracing studies have confirmed that the first and massive response to liver injury is the proliferation of mature hepatocytes 26-29 . Yet, consistent with an alternative BEC-mediated liver repair identified in human liver diseases, the BEC origin of de novo hepatocytes has been demonstrated in mouse models in which hepatocyte proliferation was significantly impaired by lack of Mdm2 30 , deficiency in β1 integrin 31 or β-catenin 32 , overexpression of p21 in hepatocytes 31 , or following long-term chronic injuries 33 17 In these studies, percentages of hepatocytes mapped from BECs averaged 15% 31,32 In the same line of evidence, the inventors' lineage tracing studies in zebrafish robustly demonstrated the BEC origin of the majority of hepatocytes after near complete ablation of hepatocytes 34,35 .

Results

VEGFR Signaling Regulates BEC-to-Hepatocyte Conversion During Liver Regeneration in Zebrafish

To investigate the molecular mechanisms underlying BEC-driven liver regeneration, the inventors had previously performed a chemical screen 46 using the zebrafish hepatocyte ablation model, Tg(fabp10a:CFP-NTR) s93 , in which metronidazole (Mtz) treatment specifically ablates virtually all nitroreductase (NTR)-expressing fabp10a + hepatocytes, thereby eliciting BEC-driven liver regeneration 35 . Through this screen, the inventors reported that suppressing VEGFR signaling with two VEGFR inhibitors, SU5416 47 and SU4312 48 , significantly reduced the size of regenerating livers but did not affect the induction of early hepatocyte markers Prox1 and HNF4α in BEC-derived liver progenitor cells (LPCs) 46 , showing normal BEC-to-LPC dedifferentiation. The inventors here sought to determine whether VEGFR signaling regulates the next step in BEC-driven liver regeneration, LPC-to-hepatocyte differentiation. To distinguish BEC-derived hepatocytes, the inventors used a Notch zebrafish reporter line, Tg(Tp1:H2B-mCherry)s939, which expresses histone 2B (H2B) and mCherry fusion proteins in BECs 35 . A long half-life of H2B-mCherry allows for tracing the lineage of BECs; thus, BECs are strong H2B-mCherry+ and BEC-derived hepatocytes are weak H2B-mCherry+ 35 . Treating Tg(fabp10a:CFP-NTR) larvae with SU5416 from ablation 18 hours (A18h) to regeneration 24 hours (R24h) significantly reduced the size of regenerating livers ( FIG. 1 a ), as reported 46 . Importantly, the SU5416 treatment greatly suppressed the expression of the hepatocyte marker Bhmt at R24h ( FIG. 1 A ), showing a defect in LPC-to-hepatocyte differentiation. This defect was confirmed with two additional hepatocyte markers, gc and f5 ( FIG. 1 B ). Complimentary to the pharmacological inhibition, the inventors genetically block VEGFR signaling using the Tg(hs:sflt1)bns80 line, which expresses a soluble form of VEGFR1 (sFlt1), a decoy receptor for VEGFA, VEGFB, and PlGF 41 , upon heat-shock 49 . sFlt1 overexpression also reduced Bhmt expression and liver size in regenerating livers at R26h ( FIG. 1 C ), phenocopying the effects of the SU5416 treatment.

›EXAMPLES · 2 of 2

Inversely, the inventors next investigated if the overactivation of VEGFR signaling could promote LPC-to-hepatocyte differentiation. The inventors used the Tg(hs:loxP-mCherry-loxP-hVEGFA) mn32 line, which expresses human VEGFA165 upon heat-shock following Cre-mediated, mCherry-loxP excision 50 , together with the Tg(Tp1:CreERT2)s959 line that expresses CreERT2 in BECs. Larvae were treated with 4-OHT from 2.5 to 3.5 days post-fertilization (dpf) to induce the Cre-mediated excision of mCherry-loxP in BECs, heat-shocked twice at A20h and A

›Tables in the description — 10

TABLE 1 — Sequences used in present disclosure SEQ ID

Name	Sequence	NO:
Human	AAGGATCCCAAGGCCCAACTCCCCGAACCACTCAGGGTCCTGTGGACAG	1
Growth	CTCACCTAGCTGCAATGGCTACAGGCTCCCGGACGTCCCTGCTCCTGGCT
Hormone 1	TTTGGCCTGCTCTGCCTGCCCTGGCTTCAAGAGGGCAGTGCCTTCCCAAC
(GH)	CATTCCCTTATCCAGGCTTTTTGACAACGCTATGCTCCGCGCCCATCGTCT
(variant 1)	GCACCAGCTGGCCTTTGACACCTACCAGGAGTTTGAAGAAGCCTATATC
NCBI Ref	CCAAAGGAACAGAAGTATTCATTCCTGCAGAACCCCCAGACCTCCCTCT
Seq:	GTTTCTCAGAGTCTATTCCGACACCCTCCAACAGGGAGGAAACACAACA
NM_000515.5	GAAATCCAACCTAGAGCTGCTCCGCATCTCCCTGCTGCTCATCCAGTCGT
	GGCTGGAGCCCGTGCAGTTCCTCAGGAGTGTCTTCGCCAACAGCCTGGT
	GTACGGCGCCTCTGACAGCAACGTCTATGACCTCCTAAAGGACCTAGAG
	GAAGGCATCCAAACGCTGATGGGGAGGCTGGAAGATGGCAGCCCCCGG
	ACTGGGCAGATCTTCAAGCAGACCTACAGCAAGTTCGACACAAACTCAC
	ACAACGATGACGCACTACTCAAGAACTACGGGCTGCTCTACTGCTTCAG
	GAAGGACATGGACAAGGTCGAGACATTCCTGCGCATCGTGCAGTGCCGC
	TCTGTGGAGGGCAGCTGTGGCTTCTAGCTGCCCGGGTGGCATCCCTGTGA
	CCCCTCCCCAGTGCCTCTCCTGGCCCTGGAAGTTGCCACTCCAGTGCCCA
	CCAGCCTTGTCCTAATAAAATTAAGTTGCATCA
Human	CAAAAAGAGAAACTGTTGGGAGAGGAATCGTATCTCCATATTTCTTCTTT	2
Epidermal	CAGCCCCAATCCAAGGGTTGTAGCTGGAACTTTCCATCAGTTCTTCCTTT
Growth Factor	CTTTTTCCTCTCTAAGCCTTTGCCTTGCTCTGTCACAGTGAAGTCAGCCAG
(EGF)	AGCAGGGCTGTTAAACTCTGTGAAATTTGTCATAAGGGTGTCAGGTATTT
(Variant 1)	CTTACTGGCTTCCAAAGAAACATAGATAAAGAAATCTTTCCTGTGGCTTC
NCBI Ref	CCTTGGCAGGCTGCATTCAGAAGGTCTCTCAGTTGAAGAAAGAGCTTGG
Seq:	AGGACAACAGCACAACAGGAGAGTAAAAGATGCCCCAGGGCTGAGGCC
NM_001963.6	TCCGCTCAGGCAGCCGCATCTGGGGTCAATCATACTCACCTTGCCCGGGC
	CATGCTCCAGCAAAATCAAGCTGTTTTCTTTTGAAAGTTCAAACTCATCA
	AGATTATGCTGCTCACTCTTATCATTCTGTTGCCAGTAGTTTCAAAATTTA
	GTTTTGTTAGTCTCTCAGCACCGCAGCACTGGAGCTGTCCTGAAGGTACT
	CTCGCAGGAAATGGGAATTCTACTTGTGTGGGTCCTGCACCCTTCTTAAT
	TTTCTCCCATGGAAATAGTATCTTTAGGATTGACACAGAAGGAACCAATT
	ATGAGCAATTGGTGGTGGATGCTGGTGTCTCAGTGATCATGGATTTTCAT
	TATAATGAGAAAAGAATCTATTGGGTGGATTTAGAAAGACAACTTTTGC
	AAAGAGTTTTTCTGAATGGGTCAAGGCAAGAGAGAGTATGTAATATAGA
	GAAAAATGTTTCTGGAATGGCAATAAATTGGATAAATGAAGAAGTTATT
	TGGTCAAATCAACAGGAAGGAATCATTACAGTAACAGATATGAAAGGA
	AATAATTCCCACATTCTTTTAAGTGCTTTAAAATATCCTGCAAATGTAGC
	AGTTGATCCAGTAGAAAGGTTTATATTTTGGTCTTCAGAGGTGGCTGGAA
	GCCTTTATAGAGCAGATCTCGATGGTGTGGGAGTGAAGGCTCTGTTGGA
	GACATCAGAGAAAATAACAGCTGTGTCATTGGATGTGCTTGATAAGCGG
	CTGTTTTGGATTCAGTACAACAGAGAAGGAAGCAATTCTCTTATTTGCTC
	CTGTGATTATGATGGAGGTTCTGTCCACATTAGTAAACATCCAACACAGC
	ATAATTTGTTTGCAATGTCCCTTTTTGGTGACCGTATCTTCTATTCAACAT
	GGAAAATGAAGACAATTTGGATAGCCAACAAACACACTGGAAAGGACA
	TGGTTAGAATTAACCTCCATTCATCATTTGTACCACTTGGTGAACTGAAA
	GTAGTGCATCCACTTGCACAACCCAAGGCAGAAGATGACACTTGGGAGC
	CTGAGCAGAAACTTTGCAAATTGAGGAAAGGAAACTGCAGCAGCACTGT
	GTGTGGGCAAGACCTCCAGTCACACTTGTGCATGTGTGCAGAGGGATAC
	GCCCTAAGTCGAGACCGGAAGTACTGTGAAGATGTTAATGAATGTGCTT
	TTTGGAATCATGGCTGTACTCTTGGGTGTAAAAACACCCCTGGATCCTAT
	TACTGCACGTGCCCTGTAGGATTTGTTCTGCTTCCTGATGGGAAACGATG
	TCATCAACTTGTTTCCTGTCCACGCAATGTGTCTGAATGCAGCCATGACT
	GTGTTCTGACATCAGAAGGTCCCTTATGTTTCTGTCCTGAAGGCTCAGTG
	CTTGAGAGAGATGGGAAAACATGTAGCGGTTGTTCCTCACCCGATAATG
	GTGGATGTAGCCAGCTCTGCGTTCCTCTTAGCCCAGTATCCTGGGAATGT
	GATTGCTTTCCTGGGTATGACCTACAACTGGATGAAAAAAGCTGTGCAG
	CTTCAGGACCACAACCATTTTTGCTGTTTGCCAATTCTCAAGATATTCGA
	CACATGCATTTTGATGGAACAGACTATGGAACTCTGCTCAGCCAGCAGA
	TGGGAATGGTTTATGCCCTAGATCATGACCCTGTGGAAAATAAGATATA
	CTTTGCCCATACAGCCCTGAAGTGGATAGAGAGAGCTAATATGGATGGT
	TCCCAGCGAGAAAGGCTTATTGAGGAAGGAGTAGATGTGCCAGAAGGTC
	TTGCTGTGGACTGGATTGGCCGTAGATTCTATTGGACAGACAGAGGGAA
	ATCTCTGATTGGAAGGAGTGATTTAAATGGGAAACGTTCCAAAATAATC
	ACTAAGGAGAACATCTCTCAACCACGAGGAATTGCTGTTCATCCAATGG
	CCAAGAGATTATTCTGGACTGATACAGGGATTAATCCACGAATTGAAAG
	TTCTTCCCTCCAAGGCCTTGGCCGTCTGGTTATAGCCAGCTCTGATCTAA
	TCTGGCCCAGTGGAATAACGATTGACTTCTTAACTGACAAGTTGTACTGG
	TGCGATGCCAAGCAGTCTGTGATTGAAATGGCCAATCTGGATGGTTCAA
	AACGCCGAAGACTTACCCAGAATGATGTAGGTCACCCATTTGCTGTAGC
	AGTGTTTGAGGATTATGTGTGGTTCTCAGATTGGGCTATGCCATCAGTAA
	TGAGAGTAAACAAGAGGACTGGCAAAGATAGAGTACGTCTCCAAGGCA
	GCATGCTGAAGCCCTCATCACTGGTTGTGGTTCATCCATTGGCAAAACCA
	GGAGCAGATCCCTGCTTATATCAAAACGGAGGCTGTGAACATATTTGCA
	AAAAGAGGCTTGGAACTGCTTGGTGTTCGTGTCGTGAAGGTTTTATGAA
	AGCCTCAGATGGGAAAACGTGTCTGGCTCTGGATGGTCATCAGCTGTTG
	GCAGGTGGTGAAGTTGATCTAAAGAACCAAGTAACACCATTGGACATCT
	TGTCCAAGACTAGAGTGTCAGAAGATAACATTACAGAATCTCAACACAT
	GCTAGTGGCTGAAATCATGGTGTCAGATCAAGATGACTGTGCTCCTGTG
	GGATGCAGCATGTATGCTCGGTGTATTTCAGAGGGAGAGGATGCCACAT
	GTCAGTGTTTGAAAGGATTTGCTGGGGATGGAAAACTATGTTCTGATATA
	GATGAATGTGAGATGGGTGTCCCAGTGTGCCCCCCTGCCTCCTCCAAGTG
	CATCAACACCGAAGGTGGTTATGTCTGCCGGTGCTCAGAAGGCTACCAA
	GGAGATGGGATTCACTGTCTTGATATTGATGAGTGCCAACTGGGGGAGC
	ACAGCTGTGGAGAGAATGCCAGCTGCACAAATACAGAGGGAGGCTATA
	CCTGCATGTGTGCTGGACGCCTGTCTGAACCAGGACTGATTTGCCCTGAC
	TCTACTCCACCCCCTCACCTCAGGGAAGATGACCACCACTATTCCGTAAG
	AAATAGTGACTCTGAATGTCCCCTGTCCCACGATGGGTACTGCCTCCATG
	ATGGTGTGTGCATGTATATTGAAGCATTGGACAAGTATGCATGCAACTGT
	GTTGTTGGCTACATCGGGGAGCGATGTCAGTACCGAGACCTGAAGTGGT
	GGGAACTGCGCCACGCTGGCCACGGGCAGCAGCAGAAGGTCATCGTGGT
	GGCTGTCTGCGTGGTGGTGCTTGTCATGCTGCTCCTCCTGAGCCTGTGGG
	GGGCCCACTACTACAGGACTCAGAAGCTGCTATCGAAAAACCCAAAGAA
	TCCTTATGAGGAGTCGAGCAGAGATGTGAGGAGTCGCAGGCCTGCTGAC
	ACTGAGGATGGGATGTCCTCTTGCCCTCAACCTTGGTTTGTGGTTATAAA
	AGAACACCAAGACCTCAAGAATGGGGGTCAACCAGTGGCTGGTGAGGA
	TGGCCAGGCAGCAGATGGGTCAATGCAACCAACTTCATGGAGGCAGGAG
	CCCCAGTTATGTGGAATGGGCACAGAGCAAGGCTGCTGGATTCCAGTAT
	CCAGTGATAAGGGCTCCTGTCCCCAGGTAATGGAGCGAAGCTTTCATAT
	GCCCTCCTATGGGACACAGACCCTTGAAGGGGGTGTCGAGAAGCCCCAT
	TCTCTCCTATCAGCTAACCCATTATGGCAACAAAGGGCCCTGGACCCACC
	ACACCAAATGGAGCTGACTCAGTGAAAACTGGAATTAAAAGGAAAGTC
	AAGAAGAATGAACTATGTCGATGCACAGTATCTTTTCTTTCAAAAGTAG
	AGCAAAACTATAGGTTTTGGTTCCACAATCTCTACGACTAATCACCTACT
	CAATGCCTGGAGACAGATACGTAGTTGTGCTTTTGTTTGCTCTTTTAAGC
	AGTCTCACTGCAGTCTTATTTCCAAGTAAGAGTACTGGGAGAATCACTAG
	GTAACTTATTAGAAACCCAAATTGGGACAACAGTGCTTTGTAAATTGTGT
	TGTCTTCAGCAGTCAATACAAATAGATTTTTGTTTTTGTTGTTCCTGCAGC
	CCCAGAAGAAATTAGGGGTTAAAGCAGACAGTCACACTGGTTTGGTCAG
	TTACAAAGTAATTTCTTTGATCTGGACAGAACATTTATATCAGTTTCATG
	AAATGATTGGAATATTACAATACCGTTAAGATACAGTGTAGGCATTTAA
	CTCCTCATTGGCGTGGTCCATGCTGATGATTTTGCAAAATGAGTTGTGAT
	GAATCAATGAAAAATGTAATTTAGAAACTGATTTCTTCAGAATTAGATG
	GCTTATTTTTTAAAATATTTGAATGAAAACATTTTATTTTTAAAATATTAC
	ACAGGAGGCTTCGGAGTTTCTTAGTCATTACTGTCCTTTTCCCCTACAGA
	ATTTTCCCTCTTGGTGTGATTGCACAGAATTTGTATGTATTTTCAGTTACA
	AGATTGTAAGTAAATTGCCTGATTTGTTTTCATTATAGACAACGATGAAT
	TTCTTCTAATTATTTAAATAAAATCACCAAAAACATAAACATTTTATTGT
	ATGCCTGATTAAGTAGTTAATTATAGTCTAAGGCAGTACTAGAGTTGAAC
	CAAAATGATTTGTCAAGCTTGCTGATGTTTCTGTTTTTCGTTTTTTTTTTTT
	TTCCGGAGAGAGGATAGGATCTCACTCTGTTATCCAGGCTGGAGTGTGC
	AATGGCACAATCATAGCTCAGTGCAGCCTCAAACTCCTGGGCTCAAGCA
	ATCCTCCTGCCTCAGCCTCCCGAGTAACTAGGACCACAGGCACAGGCCA
	CCATGCCTGGCTAAGGTTTTTATTTTTATTTTTTGTAGACATGGGGATCAC
	ACAATGTTGCCCAGGCTGGTCTTGAACTCCTGGCCTCAAGCAAGGTCGTG
	CTGGTAATTTTGCAAAATGAATTGTGATTGACTTTCAGCCTCCCAACGTA
	TTAGATTATAGGCATTAGCCATGGTGCCCAGCCTTGTAACTTTTAAAAAA
	ATTTTTTAATCTACAACTCTGTAGATTAAAATTTCACATGGTGTTCTAATT
	AAATATTTTTCTTGCAGCCAAGATATTGTTACTACAGATAACACAACCTG
	ATATGGTAACTTTAAATTTTGGGGGCTTTGAATCATTCAGTTTATGCATT
	AACTAGTCCCTTTGTTTATCTTTCATTTCTCAACCCCTTGTACTTTGGTGA
	TACCAGACATCAGAATAAAAAGAAATTGAAGTACCTGTTTTCAAATGGA
	TACTTTATAGGAATTTTGGTAAAGATTTGGTGATGGGAGGATGACTTGAG
	GTTTGTGGATATTAGTTAATTATTCAGTATGATACCTCACCCAGCTAATT
	TAGATTTTTCTATATTCGGTTTTGCTTTCATTGACAATATCCTGGAGGATC
	AGAAGACTTGTCTATTTCTGCTGAGTCACTGGCCTCAGAAAAATAATAAC
	CATAATTTCCCCCAAGGTTTTCTTTACCTAAGTGTGAATATTTTTTCTTCC
	TCCAAAAGCTCACTTTTGGGTTTAGATTAAATTTTTGTATTTTAGCACCTT
	TTTCTTTTAGGGGTTCAATGATGACAAAAGAAATGACATGAGAACACGG
	CTACCCATAACATACCATTATCTTTGTACCAGAAAAATCCTTGTTTCCTTC
	TTAATGACTCTGGTACCTTAGAAACTGGGACCCTGCTAAGTCCTTGACTA
	GGCTATCTACCAGCTCCTGGTCGGATTAAAGAAAAAACACACTTTGTGTT
	TTTTAATCACCAAGGCACCCTGCAGAGATATCTTCTTCTTGCAACTTCAC
	ATCTTTATCAGTAATGTCCTCTTTCCTTTAAAAATTCAAGTTTTAAGAACA
	GCATTTTCATGTAAAAACTTGATTTGTGTTTTTTCCAGACTGAATACTTTT
	CCTCCCTAACTCTCATCGTCTCATTGCGCGCAACGCCTGATTGAGCTTCT
	GTTTGACTAAATATCACCTACTATGTAAAAAATGAGCATATTGGCCTCTT
	TTCTAGCATCTAATAAAGGCTTAATACACTGTA
Human	AGGCACTGACTCCGAACAGGATTCTTTCACCCAGGCATCTCCTCCAGAG	3
hepatocyte	GGATCCGCCAGCCCGTCCAGCAGCACCATGTGGGTGACCAAACTCCTGC
growth factor	CAGCCCTGCTGCTGCAGCATGTCCTCCTGCATCTCCTCCTGCTCCCCATC
(HGF)	GCCATCCCCTATGCAGAGGGACAAAGGAAAAGAAGAAATACAATTCAT
Variant 1	GAATTCAAAAAATCAGCAAAGACTACCCTAATCAAAATAGATCCAGCAC
NCBI Ref	TGAAGATAAAAACCAAAAAAGTGAATACTGCAGACCAATGTGCTAATAG
Seq:	ATGTACTAGGAATAAAGGACTTCCATTCACTTGCAAGGCTTTTGTTTTTG
NM_000601.6	ATAAAGCAAGAAAACAATGCCTCTGGTTCCCCTTCAATAGCATGTCAAG
	TGGAGTGAAAAAAGAATTTGGCCATGAATTTGACCTCTATGAAAACAAA
	GACTACATTAGAAACTGCATCATTGGTAAAGGACGCAGCTACAAGGGAA
	CAGTATCTATCACTAAGAGTGGCATCAAATGTCAGCCCTGGAGTTCCATG
	ATACCACACGAACACAGCTTTTTGCCTTCGAGCTATCGGGGTAAAGACCT
	ACAGGAAAACTACTGTCGAAATCCTCGAGGGGAAGAAGGGGGACCCTG
	GTGTTTCACAAGCAATCCAGAGGTACGCTACGAAGTCTGTGACATTCCTC
	AGTGTTCAGAAGTTGAATGCATGACCTGCAATGGGGAGAGTTATCGAGG
	TCTCATGGATCATACAGAATCAGGCAAGATTTGTCAGCGCTGGGATCAT
	CAGACACCACACCGGCACAAATTCTTGCCTGAAAGATATCCCGACAAGG
	GCTTTGATGATAATTATTGCCGCAATCCCGATGGCCAGCCGAGGCCATG
	GTGCTATACTCTTGACCCTCACACCCGCTGGGAGTACTGTGCAATTAAAA
	CATGCGCTGACAATACTATGAATGACACTGATGTTCCTTTGGAAACAACT
	GAATGCATCCAAGGTCAAGGAGAAGGCTACAGGGGCACTGTCAATACCA
	TTTGGAATGGAATTCCATGTCAGCGTTGGGATTCTCAGTATCCTCACGAG
	CATGACATGACTCCTGAAAATTTCAAGTGCAAGGACCTACGAGAAAATT
	ACTGCCGAAATCCAGATGGGTCTGAATCACCCTGGTGTTTTACCACTGAT
	CCAAACATCCGAGTTGGCTACTGCTCCCAAATTCCAAACTGTGATATGTC
	ACATGGACAAGATTGTTATCGTGGGAATGGCAAAAATTATATGGGCAAC
	TTATCCCAAACAAGATCTGGACTAACATGTTCAATGTGGGACAAGAACA
	TGGAAGACTTACATCGTCATATCTTCTGGGAACCAGATGCAAGTAAGCT
	GAATGAGAATTACTGCCGAAATCCAGATGATGATGCTCATGGACCCTGG
	TGCTACACGGGAAATCCACTCATTCCTTGGGATTATTGCCCTATTTCTCG
	TTGTGAAGGTGATACCACACCTACAATAGTCAATTTAGACCATCCCGTAA
	TATCTTGTGCCAAAACGAAACAATTGCGAGTTGTAAATGGGATTCCAAC
	ACGAACAAACATAGGATGGATGGTTAGTTTGAGATACAGAAATAAACAT
	ATCTGCGGAGGATCATTGATAAAGGAGAGTTGGGTTCTTACTGCACGAC
	AGTGTTTCCCTTCTCGAGACTTGAAAGATTATGAAGCTTGGCTTGGAATT
	CATGATGTCCACGGAAGAGGAGATGAGAAATGCAAACAGGTTCTCAATG
	TTTCCCAGCTGGTATATGGCCCTGAAGGATCAGATCTGGTTTTAATGAAG
	CTTGCCAGGCCTGCTGTCCTGGATGATTTTGTTAGTACGATTGATTTACCT
	AATTATGGATGCACAATTCCTGAAAAGACCAGTTGCAGTGTTTATGGCTG
	GGGCTACACTGGATTGATCAACTATGATGGCCTATTACGAGTGGCACAT
	CTCTATATAATGGGAAATGAGAAATGCAGCCAGCATCATCGAGGGAAGG
	TGACTCTGAATGAGTCTGAAATATGTGCTGGGGCTGAAAAGATTGGATC
	AGGACCATGTGAGGGGGATTATGGTGGCCCACTTGTTTGTGAGCAACAT
	AAAATGAGAATGGTTCTTGGTGTCATTGTTCCTGGTCGTGGATGTGCCAT
	TCCAAATCGTCCTGGTATTTTTGTCCGAGTAGCATATTATGCAAAATGGA
	TACACAAAATTATTTTAACATATAAGGTACCACAGTCATAGCTGAAGTA
	AGTGTGTCTGAAGCACCCACCAATACAACTGTCTTTTACATGAAGATTTC
	AGAGAATGTGGAATTTAAAATGTCACTTACAACAATCCTAAGACAACTA
	CTGGAGAGTCATGTTTGTTGAAATTCTCATTAATGTTTATGGGTGTTTTCT
	GTTGTTTTGTTTGTCAGTGTTATTTTGTCAATGTTGAAGTGAATTAAGGTA
	CATGCAAGTGTAATAACATATCTCCTGAAGATACTTGAATGGATTAAAA
	AAACACACAGGTATATTTGCTGGATGATAAAGATTTCATGGGAAAAAAA
	ATCAATTAATCTGTCTAAGCTGCTTTCTGATGTTGGTTTCTTAATAATGAG
	TAAACCACAAATTAAATGTTATTTTAACCTCACCAAAACAATTTATACCT
	TGTGTCCCTAAATTGTAGCCCTATATTAAATTATATTACATTTCATATGCT
	ATATGTTATAGTTCATTCATTTCTCTTCACCATGTATCCTGCAATACTGGT
	ACACGAACACACTTTTTACAAAACCACATACCCATGTACACATGCCTAG
	GTACACATGTGCATGCACTACAGTTTAAATTATGGTGTACCTAATGTAAC
	CCCTAAATATTTTAGAAGTATGTACCTATAGTTTTACCTCAAAAAAACCA
	GAAATCTCTAAAGACCAGTAGAAATATTAAAAAATGATGCAAGATCAAA
	ATGATTAGCTAATTCTCCATACATAATCTGCAGATGATCTTCTTTGGTTG
	GCATTTCAGGTGTGGCCATCACCCAGAGTTAAATAACACCTAATCTAGGT
	GTTTACATGTATTCATTATCCTAGTTATTTCATGTAGTTTCTAATTCTTAA
	AGGAAAGAGGGTAATAGTTCTATTTGTGTAATTTGTTTCCTCCAAACTTA
	AGGCCACTTATTTACACAAGATATTTGTAGATCTATTTTCCTAAAGCATT
	TCTTAAGTGCTCAGATCAGTATCTAATTGAAGAAGTTTAAAAGTGTTTTG
	GTCATTAAAAATGTACTTAAATAGGTTAAATCTAAGCCTTGCTGCTGTGA
	TTGGCTTCTAGCTCACTGCCTTTAAATTTTAAAAAATTTAAGAGGAAAAT
	TTCCAAGTCTCCAAAGTTTTATAAATACCCTTCATCAAGTCATGCATTAA
	AGTATATATTGGAGAAAAAAATAAAAATACTTTTCTCAACCTGGAAGAT
	TTTAGCCTAATAAAGCTTTTTTGAAGTAAAAGACAACTTGTAAAAGGAA
	AGAAACTAGTTTGTCTCAACTCTGTATTCATTTATTTTTTTTTTGAAGTAG
	AGTGGAATCTGTTGAATCAGATATTTTATCAAGATATGTTTATTTTTTCTT
	ATTTCATTTTACAAAGTTCACTCCTAATGCCATATGTAACAGACATTTAA
	ATTTTGTGTTCTGTATAACAGCCAAATTATCATATTTATCATTGTATTTGT
	CATGCTTAGCTAAAGATCATGTATTTGTTGAGAAATAGAATAACAAAAA
	GTAATAGGATAGGCTTTGAATTTTTGCAGAAATCTTCCTGTACAAAACAC
	CTTTAAAAATAATTTTTTGAATGGTGTGAATCCAGTAGTCCCATTTCTCT
	GACTTAGTTTTCTTGAGTGATTTTTATCAAGGCCAAGTCCCCAAACAATT
	CCCTACCAGCTCTTTAGAGTACTGTTCAATCTGGACTAAAATGGTTTTAA
	GTTTATGGAGAGCTTAGTCCACAGAATATAGGGCGGCGAGTCCAGAAAT
	GCTTATACAATTTTTTTTTCATAATAAGATATGTGCTGGCATCAAGAAAC
	TTAAAGTGGAAGCAAAAAGACATCCAACTAGTTGCTGGTCTCTATCATCT
	TATCTGATGGTATTTCTATTTTCCTTATATAATACACCATTTTAGTAAGAA
	CTCCTAGAAATTTCAAGAGCATATTGCCAAAATATAAAGTATATTTCATA
	GTTTCTTCTGGCTGAACCAGTGAAATTTTATTATTGCATATTAATGATATT
	TGTAAAACTTTTATAAAAATTGTCATAATTTTAAATACTCACATTTTAAA
	AATACTTCTTTAATGACTCTTCCTCTAAATTTCCTGGAAATACAGATAAA
	GATTAGCTAGATACAAGATACAGCTAAGTATTTAGACATTTTGAGGCTA
	GTATTTTTCATTTTATTAAAGGCTAAAAACAATACCACCAATAAATCATC
	AAACAAACCGTACAAAGTAATTCTCTCTTTGGGAGGCTCCTTTCGTGATA
	GAGGGACATGGGTGGAATTGACAATGAAACTTAGATGAACAAGGTCCAT
	GTTATTTTAGGTGGTAGAACAGGGTAGAGTCATGTCATTATTTGCTGGTG
	GAAGACACTATTTACCAGGTGTTCTTTGCTGAATAAATCATTAAACATTT
	TTAAAAATCCAACAATCCACTTTATTTTGTGTCATTGACAAAAGGATCTT
	TTAAATCAGAAGGTTTCAATGCAATTTTTGGTTTGGCTGTTTGAATAATG
	GTTATGTACTGTTATAATTGTAGACATTTTCTCACGTCTACCAGGAATTG
	AAGTGTAAAACTAAAATATTTTTCATAATGCCTCTGCCGTGCAGAAGGA
	ATGATAATCCTTTTGTATACTTCTTTAATTTTATTGTAAAATGTGTAATGA
	CTTTTACCTATATGCTGTGGGCAGGTCCTCAGTAAAATCTATTGAGTCAA
	TTTCTAGTATTAACAGGCTTTTGCTTGCTATCTAAGTGTTTCAAATTATGG
	GAAGTGTGAGACACTGGAAGGCAAGAAAATTAACAATAATGGCATGTG
	ATAGCAAAATTGTATTTCACTTATTCCTGTGAATATTTCTTGTTGGTACCA
	ATGGTACTGTACAAAGTGAATGTTATAGCCACAACATTCTCTTGAAAAG
	AACACTGTCAAGAAGTGGGAAATTGCTGTCAGGCATTTCATTGTTGTTTT
	TAAACTTTTTTAAAAGAAATACTGGTTTTGCAATATAGAGATCATGTGGT
	AAAGAATTTTAATAAGATCTTATACTAAAAAGCCTTAAATCAATTTATTG
	AGATTCAAAAAATACTATTATAATTAATTACATCCCATACATATAGGCAA
	ACTCATTTAAAAAATAAAACTAATTTTGGTAAAAGTACATGGCCTTTGTT
	TTTAAAATACATAATTTTAAAATAAATCACTTGTCATGATAAAGTCCAAA
	AAGAAGTTATCATTCAACATTCAACTAAGGTTGGAGCTAAGAATTTACT
	AATACAAAAAAAGTTAAAATTTTTTGGACCATATATATCTTGACAGTGTA
	ACTTTTAAGTAGGTTCATTTCCATTTGCACAGAAAGTTTCTGTCTTTAGG
	AAACTGAAAATGAAATACTGTGGATGCTATGACTGTTTGTCTTGTATGTA
	AATAGGAAATTAATAAGCTGCCTATTGAGTGGTATAGCTGTATGCTTACC
	CAAAAAAGGGAACACTGTGGTTATGACTTGTATTATAAACTTTCTGTAGT
	TAATAAAGTTGTTATTTTTATAACCATGATTATATTATTATTATTAATAAA
	ATATTTTATCAAAA
Mouse Cyclin-	AGTGCAGGGTGGTGGAGACCTGATGATACCCAACTACCAGCTGTGGGGT	4
dependent	GAGGAGGAGCATGAATGGAGACAGAGACCCCAGATAATTAAGGACGTC
kinase	CCACTTTGCCAGCAGAATAAAAGGTGCCACAGGCACCATGTCCAATCCT
inhibitor	GGTGATGTCCGACCTGTTCCGCACAGGAGCAAAGTGTGCCGTTGTCTCTT
1A/Senescence	CGGTCCCGTGGACAGTGAGCAGTTGCGCCGTGATTGCGATGCGCTCATG
gene (p21)	GCGGGCTGTCTCCAGGAGGCCCGAGAACGGTGGAACTTTGACTTCGTCA
(Cdkn1a)	CGGAGACGCCGCTGGAGGGCAACTTCGTCTGGGAGCGCGTTCGGAGCCT
(Variant 2)	AGGGCTGCCCAAGGTCTACCTGAGCCCTGGGTCCCGCAGCCGTGACGAC
NCBI Ref	CTGGGAGGGGACAAGAGGCCCAGTACTTCCTCTGCCCTGCTGCAGGGGC
Seq:	CAGCTCCGGAGGACCACGTGGCCTTGTCGCTGTCTTGCACTCTGGTGTCT
NM_001111099.2	GAGCGGCCTGAAGATTCCCCGGGTGGGCCCGGAACATCTCAGGGCCGAA
	AACGGAGGCAGACCAGCCTGACAGATTTCTATCACTCCAAGCGCAGATT
	GGTCTTCTGCAAGAGAAAACCCTGAAGTGCCCACGGGAGCCCCGCCCTC
	TTCTGCTGTGGGTCAGGAGGCCTCTTCCCCATCTTCGGCCTTAGCCCTCA
	CTCTGTGTGTCTTAATTATTATTTGTGTTTTAATTTAAACGTCTCCTGTAT
	ATACGCTGCCTGCCCTCTCCCAGTCTCCAAACTTAAAGTTATTTAAAAAA
	AGAACAAAACAAAACAAAAAAAACCAAAACAAAACAAACCTAAATTAG
	TAGGACGGTAGGGCCCTTAGTGTGGGGGATTTCTATTATGTAGATTATTA
	TTATTTAAGCCCCTCCCAACCCAAGCTCTGTGTTTCCTATACCGGAGGAA
	CAGTCCTACTGATATCAACCCATCTGCATCCGTTTCACCCAACCCCCCTC
	CCCCCATTCCCTGCCTGGTTCCTTGCCACTTCTTACCTGGGGGTGATCCTC
	AGACCTGAATAGCACTTTGGAAAAATGAGTAGGACTTTGGGGTCTCCTT
	GTCACCTCTAAGGCCAGCTAGGATGACAGTGAAGCAGTCACAGCCTAGA
	ACAGGGATGGCAGTTAGGACTCAACCGTAATATCCCGACTCTTGACATT
	GCTCAGACCTGTGAAGACAGGAATGGTCCCCACTCTGGATCCCCTTTGCC
	ACTCCTGGGGAGCCCACCTCTCCTGTGGGTCTCTGCCAGCTGCCCCTCTA
	TTTTGGAGGGTTAATCTGGTGATCTGCTGCTCTTTTCCCCCACCCCATACT
	TCCCCTTCTGCAGGTCGGCAGGAGGCATATCTAGGCACTTGCCCCACAGC
	TCAGTGGACTGGAAGGGAATGTATATGCAGGGTACACTAAGTGGGATTC
	CCTGGTCTTACCTTAGGCAGCTCCAGTGGCAACCCCCTGCATTGTGGGTC
	TAGGGTGGGTCCTTGGTGGTGAGACAGGCCTCCCAGAGCATTCTATGGT
	GTGTGGTGGTGGGGGTGGGCTTATCTGGGATGGGGACCCCAGTTGGGGT
	TCTCAGTGACTTCTCCCATTTCTTAGTAGCAGTTGTACAAGGAGCCAGGC
	CAAGATGGTGTCTTGGGGGCTAAGGGAGCTCACAGGACACTGAGCAATG
	GCTGATCCTTTCTCAGTGTTGAATACCGTGGGTGTCAAAGCACTTAGTGG
	GTCTGACTCCAGCCCCAAACATCCCTGTTTCTGTAACATCCTGGTCTGGA
	CTGTCTACCCTTAGCCCGCACCCCAAGAACATGTATTGTGGCTCCCTCCC
	TGTCTCCACTCAGATTGTAAGCGTCTCACGAGAAGGGACAGCACCCTGC
	ATTGTCCCGAGTCCTCACACCCGACCCCAAAGCTGGTGCTCAATAAATAC
	TTCTCGATGATT
Human	GCGGAGGCTTGGGGCAGCCGGGTAGCTCGGAGGTCGTGGCGCTGGGGGC	5
vascular	TAGCACCAGCGCTCTGTCGGGAGGCGCAGCGGTTAGGTGGACCGGTCAG
endothelial	CGGACTCACCGGCCAGGGCGCTCGGTGCTGGAATTTGATATTCATTGATC
growth factor	CGGGTTTTATCCCTCTTCTTTTTTCTTAAACATTTTTTTTTAAAACTGTATT
A (VEGFA)	GTTTCTCGTTTTAATTTATTTTTGCTTGCCATTCCCCACTTGAATCGGGCC
(variant 5)	GACGGCTTGGGGAGATTGCTCTACTTCCCCAAATCACTGTGGATTTTGGA
NCBI Ref	AACCAGCAGAAAGAGGAAAGAGGTAGCAAGAGCTCCAGAGAGAAGTCG
Seq:	AGGAAGAGAGAGACGGGGTCAGAGAGAGCGCGCGGGCGTGCGAGCAGC
NM_001025369.3	GAAAGCGACAGGGGCAAAGTGAGTGACCTGCTTTTGGGGGTGACCGCCG
	GAGCGCGGCGTGAGCCCTCCCCCTTGGGATCCCGCAGCTGACCAGTCGC
	GCTGACGGACAGACAGACAGACACCGCCCCCAGCCCCAGCTACCACCTC
	CTCCCCGGCCGGCGGCGGACAGTGGACGCGGCGGCGAGCCGCGGGCAG
	GGGCCGGAGCCCGCGCCCGGAGGCGGGGTGGAGGGGGTCGGGGCTCGC
	GGCGTCGCACTGAAACTTTTCGTCCAACTTCTGGGCTGTTCTCGCTTCGG
	AGGAGCCGTGGTCCGCGCGGGGGAAGCCGAGCCGAGCGGAGCCGCGAG
	AAGTGCTAGCTCGGGCCGGGAGGAGCCGCAGCCGGAGGAGGGGGAGGA
	GGAAGAAGAGAAGGAAGAGGAGAGGGGGCCGCAGTGGCGACTCGGCGC
	TCGGAAGCCGGGCTCATGGACGGGTGAGGCGGCGGTGTGCGCAGACAGT
	GCTCCAGCCGCGCGCGCTCCCCAGGCCCTGGCCCGGGCCTCGGGCCGGG
	GAGGAAGAGTAGCTCGCCGAGGCGCCGAGGAGAGCGGGCCGCCCCACA
	GCCCGAGCCGGAGAGGGAGCGCGAGCCGCGCCGGCCCCGGTCGGGCCT
	CCGAAACCATGAACTTTCTGCTGTCTTGGGTGCATTGGAGCCTTGCCTTG
	CTGCTCTACCTCCACCATGCCAAGTGGTCCCAGGCTGCACCCATGGCAGA
	AGGAGGAGGGCAGAATCATCACGAAGTGGTGAAGTTCATGGATGTCTAT
	CAGCGCAGCTACTGCCATCCAATCGAGACCCTGGTGGACATCTTCCAGG
	AGTACCCTGATGAGATCGAGTACATCTTCAAGCCATCCTGTGTGCCCCTG
	ATGCGATGCGGGGGCTGCTGCAATGACGAGGGCCTGGAGTGTGTGCCCA
	CTGAGGAGTCCAACATCACCATGCAGATTATGCGGATCAAACCTCACCA
	AGGCCAGCACATAGGAGAGATGAGCTTCCTACAGCACAACAAATGTGAA
	TGCAGACCAAAGAAAGATAGAGCAAGACAAGAAAATCCCTGTGGGCCT
	TGCTCAGAGCGGAGAAAGCATTTGTTTGTACAAGATCCGCAGACGTGTA
	AATGTTCCTGCAAAAACACAGACTCGCGTTGCAAGATGTGACAAGCCGA
	GGCGGTGAGCCGGGCAGGAGGAAGGAGCCTCCCTCAGGGTTTCGGGAA
	CCAGATCTCTCACCAGGAAAGACTGATACAGAACGATCGATACAGAAAC
	CACGCTGCCGCCACCACACCATCACCATCGACAGAACAGTCCTTAATCC
	AGAAACCTGAAATGAAGGAAGAGGAGACTCTGCGCAGAGCACTTTGGG
	TCCGGAGGGCGAGACTCCGGCGGAAGCATTCCCGGGCGGGTGACCCAGC
	ACGGTCCCTCTTGGAATTGGATTCGCCATTTTATTTTTCTTGCTGCTAAAT
	CACCGAGCCCGGAAGATTAGAGAGTTTTATTTCTGGGATTCCTGTAGACA
	CACCCACCCACATACATACATTTATATATATATATATTATATATATATAA
	AAATAAATATCTCTATTTTATATATATAAAATATATATATTCTTTTTTTAA
	ATTAACAGTGCTAATGTTATTGGTGTCTTCACTGGATGTATTTGACTGCT
	GTGGACTTGAGTTGGGAGGGGAATGTTCCCACTCAGATCCTGACAGGGA
	AGAGGAGGAGATGAGAGACTCTGGCATGATCTTTTTTTTGTCCCACTTGG
	TGGGGCCAGGGTCCTCTCCCCTGCCCAGGAATGTGCAAGGCCAGGGCAT
	GGGGGCAAATATGACCCAGTTTTGGGAACACCGACAAACCCAGCCCTGG
	CGCTGAGCCTCTCTACCCCAGGTCAGACGGACAGAAAGACAGATCACAG
	GTACAGGGATGAGGACACCGGCTCTGACCAGGAGTTTGGGGAGCTTCAG
	GACATTGCTGTGCTTTGGGGATTCCCTCCACATGCTGCACGCGCATCTCG
	CCCCCAGGGGCACTGCCTGGAAGATTCAGGAGCCTGGGCGGCCTTCGCT
	TACTCTCACCTGCTTCTGAGTTGCCCAGGAGACCACTGGCAGATGTCCCG
	GCGAAGAGAAGAGACACATTGTTGGAAGAAGCAGCCCATGACAGCTCC
	CCTTCCTGGGACTCGCCCTCATCCTCTTCCTGCTCCCCTTCCTGGGGTGCA
	GCCTAAAAGGACCTATGTCCTCACACCATTGAAACCACTAGTTCTGTCCC
	CCCAGGAGACCTGGTTGTGTGTGTGTGAGTGGTTGACCTTCCTCCATCCC
	CTGGTCCTTCCCTTCCCTTCCCGAGGCACAGAGAGACAGGGCAGGATCC
	ACGTGCCCATTGTGGAGGCAGAGAAAAGAGAAAGTGTTTTATATACGGT
	ACTTATTTAATATCCCTTTTTAATTAGAAATTAAAACAGTTAATTTAATTA
	AAGAGTAGGGTTTTTTTTCAGTATTCTTGGTTAATATTTAATTTCAACTAT
	TTATGAGATGTATCTTTTGCTCTCTCTTGCTCTCTTATTTGTACCGGTTTTT
	GTATATAAAATTCATGTTTCCAATCTCTCTCTCCCTGATCGGTGACAGTC
	ACTAGCTTATCTTGAACAGATATTTAATTTTGCTAACACTCAGCTCTGCC
	CTCCCCGATCCCCTGGCTCCCCAGCACACATTCCTTTGAAATAAGGTTTC
	AATATACATCTACATACTATATATATATTTGGCAACTTGTATTTGTGTGT
	ATATATATATATATATGTTTATGTATATATGTGATTCTGATAAAATAGAC
	ATTGCTATTCTGTTTTTTATATGTAAAAACAAAACAAGAAAAAATAGAG
	AATTCTACATACTAAATCTCTCTCCTTTTTTAATTTTAATATTTGTTATCA
	TTTATTTATTGGTGCTACTGTTTATCCGTAATAATTGTGGGGAAAAGATA
	TTAACATCACGTCTTTGTCTCTAGTGCAGTTTTTCGAGATATTCCGTAGTA
	CATATTTATTTTTAAACAACGACAAAGAAATACAGATATATCTTAAAAA
	AAAAAAAGCATTTTGTATTAAAGAATTTAATTCTGATCTCAAA
Mouse Insulin	ACTCGATAACTTTGCCAGAAGAGGGAGAGAGAGAGAAGGCGAATGTTC	6
Growth Factor	CCCCAGCTGTTTCCTGTCTACAGTGTCTGTGTTTTGTAGATAAATGTGAG
(IGF-1)	GATTTTCTCTAAATCCCTCTTCTGCTTGCTAAATCTCACTGTCACTGCTAA
Variant 4	ATTCAGAGCAGATAGAGCCTGCGCAATGGAATAAAGTCCTCAAAATTGA
NCBI Ref	AATGTGACATTGCTCTAACATCTCCCATCTCTCTGGATTTCTTTTTCGCCT
Seq:	CATTATCCCTGCCCACCAATTCATTTCCAGACTTTGTACTTCAGAAGCGA
NM_001111275.2	TGGGGAAAATCAGCAGCCTTCCAACTCAATTATTTAAGATCTGCCTCTGT
	GACTTCTTGAAGATAAAGATACACATCATGTCGTCTTCACACCTCTTCTA
	CCTGGCGCTCTGCTTGCTCACCTTCACCAGCTCCACCACAGCTGGACCAG
	AGACCCTTTGCGGGGCTGAGCTGGTGGATGCTCTTCAGTTCGTGTGTGGA
	CCGAGGGGCTTTTACTTCAACAAGCCCACAGGCTATGGCTCCAGCATTCG
	GAGGGCACCTCAGACAGGCATTGTGGATGAGTGTTGCTTCCGGAGCTGT
	GATCTGAGGAGACTGGAGATGTACTGTGCCCCACTGAAGCCTACAAAAG
	CAGCCCGCTCTATCCGTGCCCAGCGCCACACTGACATGCCCAAGACTCA
	GAAGGAAGTACATTTGAAGAACACAAGTAGAGGAAGTGCAGGAAACAA
	GACCTACAGAATGTAGGAGGAGCCTCCCACGGAGCAGAAAATGCCACAT
	CACCGCAGGATCCTTTGCTGCTTGAGCAACCTGCAAAACATCGAAACAC
	CTACCAAATAACAATAATAAGTCCAATAACATTACAAAGATGGGCATTT
	CCCCCAATGAAATATACAAGTAAACATTCCAACATCGTCTTTAGGAGTGT
	TTGTTTAAAAAGCTTTGCACCTTGCAAAAGTGGTCCTGGCGTGGGTAGAT
	TGCTGTTGGTCCTTTATCAATAACATTCTATAGAGAAAAAAAATATATAT
	ATAACTATATCTCCTAGTCCCTGCCTCTAAAGAGCCGAAAATGCATGGAT
	GTTGTAGAGATCCAGTTGCTCTAAGTTTCTCTCTGAATTTTGGCTGCTGA
	AGCCATTCATTTAGCAACTGTGTAGAGGTGGTTTATGAATGGTTCCCTTA
	TCTTCACCTCTTCCCACGTAGCTCAAGCTGCTTGTTTTACAGAGTCTAATC
	ATCTTGTCTAGCTGCATTAGACACACCCTTTCCTAACACTTGTATTTGTTG
	AATTTGGCCTCCTTAAGAGCAATAGCAAATAAGTAGTCAAGTGGCCTAC
	CAAGTTTTAACGTACCTGACTCCATCTGTGGCATTTGTACCAAATATAAG
	TTGAATGCATTTATTTTAGACACAAAGCTTTATTTTTTTTGACATTGTGTT
	TCAAGAAAAAAAATAGAATAACAATAACTACAACTTTGAGGCCAATCAT
	TTTTAGGTGTGTGTTTGAAGCATAGAACGTCTCTTAAACTCTCAATGGTT
	TCTTCAAATGATAAGTTAGTATGTAACCTAAGTATAGCAGTTTCTCTCTT
	TTTTATTTTTTTCCATATAGAGCACTATGTAAAGTTAGTATATCAATAATA
	CAGGAAATATCAAACAGTATGTAAAACTCTGTTGTTGTTGTTTTTTAGTA
	CAATGGTGCTATTTTGTAGTTTGTTATATGAAAGAATCTAGTCAACACAG
	TAAAAGGAGAAAGCAAAGCAAAAACAACAAACGAAAGCCTGGAGCCTA
	AGATGACAAAACGAGGAAGGGAACTGAAAAAAAAAATCCTTCCTCTTG
	GGAGATGCAAAGGCCTCCCCAATTATGCCTTCCAAGAAGAACTTAAGAT
	ATAGAGTCCATTAAGACGCACTTACTTGTCAAGTCCAGAGAGGAAGCTA
	TGGAGTGGGAAAAGCAAGAGGCTAGGGATTTGGGAGTCCTGGTTTCTTT
	TTAATCACTGAAGAAGTAAGTATTTGCAACCTGGGTCACACAAACTCAC
	CACCCTGTGACCTCAGTCAAATCACTCCACCTCTCGGTGCCTCAGTTTTC
	CTCATCTGCAAAATGGGGGCAATATGTCATCTACCTACCTCAAAGGGGT
	GGTATGAAGATTAAAAAGTAGACCTTCAGATTTTTGTTCTGGGTTTCCAG
	GAGGGTGCAACATCAGAACCCTTGAATTGCTAGGATGCAAGGAATTCTG
	TAAATAACCCACTAACAATGTAGCTCCAAGGATCATTCATCTGTCACTGG
	GATGCCACCACAATATCCAAGTTCTTATTGGTGAAGCTGTGCAACTAATT
	AGTGACAAGCTAAGGACTCAGTCTCCCCAGCATGTCACACGGCAGGAGA
	CATTTGATTTGCAGTTTTATTTAACTTCTGCATTTGAGCTTATGACTATAA
	AGACTAGTGAAAAGAAGGGAGAGAGGAGAAAGAAGATCCTTGCCAAGT
	AAAGGGTAATTAATTATTATTCCATTTATCCACTCTCATTAAAGGGTAAT
	TAATTATTCCATGTATCCACTCTCATTAATCCTTCCAGTCACTTAGTATCT
	AGAAATAACTCTAACATTGTCAATGAGACTCTACTCAGTTTGCCAAACAC
	AATTCTCCTTCCCCATAGCATATGAAAAAAAGGCGCTGACATTCTTAAAT
	TTTGAAATAGTATCTATTACAATCACAGGTTGCTGTAGCAGATGTAGTCT
	TGCCCTTGTTTGTACATGCATGTATTTTTTTTTTAATTTTATGAAAATGTG
	CTAGCAAGAATTGCTACTTGAGGGGCAAAATTCTTCCTTCTCAAGCCTGA
	GGTTCTCCCTAGTGTCTGCTTAGAAGGAAGGATCCAGCTTCCTGGAAATG
	TGTTGGATGCATTCAACTGGGCATTGCTAACCAAAAACATTTAGAAAAA
	TGTTCTCTATGTATATAGCAAGATTGTCTCCCTCTTTTAAAAACAAAATC
	CAATATTCACATCTTATTACCTACAACCTTGATTCTCTATTGCAAGCTTCC
	TTAATATTCTTATAAAATGTATTAAGAAAAACAAAAAGGACACCTTTAG
	CTCTCCTTCCGCCAGGTTGCCTCTAGAATCTCTGGGGAAATGCAGAAGGT
	GCTGTTGAGTAAAGCCCTCAGAAGGATTGGATTTAGGAACATCAGGCAC
	GCTGTACATCCCCTGATTACTGTAGAAATGTAAATGGAATAAGAGGTCA
	GCTGACCATCCACCTGCTTCCCCAGAAGGATACAGGGAAAAGTTAGGCC
	CTCACACACCCTGGGTGACACTTCTGACTTCTAGTTCTTGTTCACAGTGT
	GTACTTTTTCAAATTGGTAATTCCCAGAAAAACACATAGGTGGCCTTCTC
	CAGATCTGTGGGCTTCCTGCCATGGTTGGATTTGGTGATTCCAAGTGTCT
	ATCACATATTTTGTTCACTTAATTCTATCCACAGTCAGAAATTCTTTCAAT
	GAGGAAAGTTTAAATATGCAATCCTTTATCCAATACCTAATTCTCTCCAA
	CTGCATCATAAATCAAGTAATAAAAATTAATTGTACTAATTAATCATAAT
	AATGTACCATTGTACTTTTAAATGAATGAACACTGCAAGACAAATCTATG
	TAAACTCTGAAAAGTAACTGATCATTATATGGTGAATCAAAATGACTCA
	AGATTGATAGAAAGGGACATTTAAAATTTTACAACTCAAAATTTTGTAG
	ACTTTGCTATGGAGGTAAATTGTTTTAGTGCCTAGAGATGGAGCGGTTTT
	AATAAATTTACAAAAGAACTATAAAGATAGGTAGGAAGGAATTTTCATT
	TGATAGGATTGTTGCTGATTTACTTACTCAATACCTAGGTCAAATGTTGA
	TCCTATTCTCCAAAGACTATCAAGTGCTTGAACATTGTAAGATGAGTCTG
	CTCCACTGAAAATGTAATACATCTCTCCATTATAATCTATTTTCCTGGGG
	TAAAAAAATCCTTTTTTTAAATATCCACCTACATATACCTACCCTACATG
	TGCATTTGCACATGCGTGCATACGCTCATGCGCCCCACCCCACACACACC
	TATTCACCCTAAGACTAAGAAGAAATCATTTCTTTGAAAGTCTTATCTTT
	CAAAAAAGGCAGCGGTGCCCCTTGAGACTCCTTCTCCTTCTTTGAATGTC
	AATGTGAAATGTGGCATGTCTGTGTACATGAAACCATCTCATACCCTATG
	GCTCCAGGGTTTCTTTATGGTTTGTGCACTTGGGAGGATGCGCAGAAGAC
	AGGATGCAGCCTGTTTTGCTTTCCCCTTTACTGTTTGGCCAGCTACGCCA
	ATGTGGTGCTATTGTTTCTTTAAGAAAGTACTTGACTAAAAAAAAAGAA
	AAAAAGAAAAAAAGAAAAGAAAAAGAAAAAAGAAAAAAAAAGAAAGC
	ATAGACCTATTTTTTTAAAGTCTGAAAACAACAGTTCTATAGTAGATGGC
	TTACTGAGATAGCATTAGATCTAGCCACCACCCTAGCCACCACCTTTCAA
	CTATGTGTCACTCACAAGTAGAATATTGTTCACCAAGTTGTGAGTTTGGG
	GGTTCAGAGACAAAGGATGGAAAAGTTTTAAAGTTAGATGGCTCAATCA
	TTTCATTGGCTCTCAAATTTAACAAAATTGGCAATACTTCACCCAATCTG
	AAGTGTTGGTCAATAACTTGAACTGGGGGCAAAAATAACTTCAGGCAAA
	TGGCAGAAGAAAATAATTAACTTACTTCTTGCTTTTTTTGTTGATTGTTTG
	GTTTCCTGTTGATTTTTGGTTTTGGTTTTGCTGTGGGTGGGTGAGTACATG
	TGTGTAAGTACGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTTCC
	ACTCAAAAACAAATACTCAGAAAGTGGAGAAAATACAACGATTTTAAGA
	GCATAGACTTACCTACTACTAGAACCAGCTTCTGTCACATCCTCTGGAGA
	AGGCACTGATTTCTTGTTTTGTAGAGGTTGCTCTTCCATCAGTGACCTGA
	AAGAGTGACCAGTCTCCTAGAGTAGACATGGATCTCATTAGGAGAAGAC
	AGAAGTATTTCCTTATGAATTGGGCTTATCTACTGACAAAGAAAGGGAA
	GAGTTTATGAGAAGTTATTGAAGAAGATGGCTAACAGTCTGTGAAGATT
	TTGTTCTGGTTTTTTTTGTTGTTGTTGTTGTTGGGTTTGGGTTTTGATTTTT
	TTTTTTTTTTTTTTACTTTATACAATCTTTATGAATGGAAATCTTAATGCTC
	AAAAAGACTTGGTCTTTTTTTCTCTTTCGTAACAGAATGGAAGATGACAA
	ACTCACATAGACTCTTTCTAGGCTGGCTAGCAAAGGTGTGGTTTGACTTA
	TTTGAATCAGACCATTTTAAATGTTCCTCTCTATTTTTAATCATAAAAGGC
	TGTCATAATTTATTAGCGTAGGCCCTTTTTGGCACTTCTCAAATGAATGA
	GCATTCCCATTCAAAGCATGGCTTTCCCCATGGTTCCAAAACATGAATGA
	TTAATATTAAGGAATTATTTACTTCAAAATACAGTAGAAGTGTGAGTCTC
	TGTTCCCATTCCCCACAAAGATCATTAAGTCCTGAATCGGGGGCGGGGG
	TGGGGCGCCTGGATACTAAGGGAATTTTTTTGTTGCTTGTTTTTTGTTTTC
	AATGCTAGTGCTTAATCCTATAGTATACAGATTTGCTTCTTGCTATTGTG
	ATATTCTGTAAGACTTTCCTGTTAGGTATTAGAAATTGATACATAAATAC
	CTTTTTTGTGTGGTTTCTATTTAAAAGGAAAGAGATAAGACTGTCTGAAC
	CTTAAATTCGTAAGGCACATGATAAAGAGATCACATTAAATAACAAGCC
	ATATCTGGTTCAATCCTTTCTTTCTTATCATTTTAAGGAAAACTTGCCCAG
	ATAAGACAGAGGCCCAGGGGACTTTTGAAACTCTCTTTGTTCCGCCAATT
	CATTTTGGCTGGTGATGGTTTTTCCCCAGTGTCTGCCTCAGAATCTTTTAG
	AGGCTGGCCAGACTAAAGACTGTCTTTTAAAACACATTTCACATGGTTCC
	TCTTAATGAATGATTACACTTATGTAGAACATGATTTTTTTTTCTCTCCAC
	TTATTTTTTTTTTCCCCATCATTGATAAGGGTTCTTAAGGAGAAGAATTCA
	TTAACAAAACTCAAGAAAGCGTACAAAAAAAAAATTCTAAATGTCACTG
	CCCAATTGAAATACGAGCTAAAATGGAAATACTTTCTCCTACTTAAAACC
	CAGACTGAATCACCTTCAAAATGACCTTTCACAATCTTTCCAATTTGCCT
	TTGTTTAAACTGTCTGGGCCTAAAAGCAAGCATTATTCATTTTCTCTTGCC
	CAAAGTGAACTTGTGTAAAGTAGGAAAATTAAAAGAAACTGCTAGAAAT
	CCCTTCCAACCAGTGGCTGACCCCTCTCACTAGCTCACAGCAAAGTCTCC
	TCTGTTGATCTATCACCTAGTCTCATTTCGTTTGAATATTTACATTGTACC
	TACTGCTAAACACTTGGCAGGAGGCTCCATCCATATCTCCTATCGGTGTC
	TCTGTATCCTTAAACCTTGCAAACATCATACAGTGTATATTAAGTTTACA
	GGAAAGCTCCAAATAGCATATCAGACCTGGTCTCTCTTTGTTAAAGATTT
	AAGGAGCTATGGGAATCTGGATTACAACGCACATTTTGCTTCATTTATTT
	TTATCACACTTTAAAGGCCAAGGGTGATGATTAACTTACAGACACTGAA
	TTGATTTCCCTACTGAAACCTGAAAGTAATATTTGGTCATTCATTGTATG
	TGTTTTACACAAAAAAAACATCTTCTATCAAATTACTCCTGATTGTATTT
	GAAGTGGTTATTCAATTCATTTATGGCAGAGCAATATCTGTCCTAATGAC
	TCTTATAAAATGTAACTAACTGAATCATTATCTTACATTTACTGTTTAGTA
	AGCATATTTTGAAATTGTATGGCTAGAGTGTCATAATAAAATGGTATATC
	TTTCTTTAGTAATTACATTAAAATTAATCATGTTTGATTAACTGGT
Human Signal	TTCCTGTAACATCGCAGCCAGTGAGCAGTGAACGAGTGTAGACAGAGCT	7
transducer and	CTGCCTCTAGCCTGGCTGCCCAAGCCCAAGCCGTTAGAAGCAGGAGCCC
activator of	CTGCGCAGTGCCTGGTCACGGAGCTGAGCTGTGTTTAGATGTGTTGGCTG
transcription	CTGCGTGGTGAAGGAAGACCCGTCTCCAGAAAAGCAATTTAGGCAAAAG
5B (stat5b)	GGATTCCGTTTGATGGCAGAGTCCCAGTGCTAGAAAGGTAGCGAAGGTG
(Variant 1)	GACAGCTTACAGTCTCAACTCATTTCGTCGTAAATGTCCTCGTAACGACA
NCBI Ref	TTGATTCTTCTACCTGGATAACCTTTTGTTTGTTTGTTTGTTTGTTTTTGTT
Seq:	TTGTTTTTCCCCTGTAACCATTTTTTTTTCTGACAAGAAAACATTTTAATT
NM_011489.3	TTCTAAGCAAGAAGCATTTTTCAAATACCATGTCTGTGACCCAAAGTAAA
	AATGGATGATAATTCATGTAAATGTGTGCAACATAGCAACCTGAACCTG
	CACGCGATTCGGGCTCTGTAGGTTGTGAACCATGGCTATGTGGATACAG
	GCTCAGCAGCTCCAGGGCGATGCCCTTCACCAGATGCAGGCCTTGTACG
	GCCAGCATTTCCCCATCGAGGTGCGACATTATTTATCACAGTGGATCGAA
	AGCCAAGCCTGGGACTCAATAGATCTTGATAATCCACAGGAGAACATTA
	AGGCCACCCAGCTCCTGGAGGGCCTGGTGCAGGAGCTGCAGAAGAAGG
	CGGAGCACCAGGTGGGGGAAGATGGGTTTTTGCTGAAGATCAAGCTGGG
	GCACTATGCCACACAGCTCCAGAGCACGTACGACCGCTGCCCCATGGAG
	CTGGTTCGCTGTATCCGGCACATTCTGTACAACGAACAGAGGCTGGTTCG
	CGAAGCCAACAACGGCAGCTCTCCAGCTGGAAGTCTTGCTGACGCCATG
	TCCCAGAAGCACCTTCAGATCAACCAAACGTTTGAGGAGCTGCGCCTGA
	TCACACAGGACACGGAGAACGAGCTGAAGAAGCTGCAGCAGACCCAAG
	AGTACTTCATCATCCAGTACCAGGAGAGCCTGCGGATCCAAGCTCAGTTT
	GCCCAGCTGGGACAGCTGAACCCCCAGGAGCGCATGAGCAGGGAGACG
	GCCCTCCAGCAGAAGCAAGTGTCCCTGGAGACCTGGCTGCAGCGAGAGG
	CACAGACACTGCAGCAGTACCGAGTGGAGCTGGCTGAGAAGCACCAGA
	AGACCCTGCAGCTGCTGCGGAAGCAGCAGACCATCATCCTGGACGACGA
	GCTGATCCAGTGGAAGCGGAGACAGCAGCTGGCCGGGAACGGGGGTCC
	CCCCGAGGGCAGCCTGGACGTGCTGCAGTCCTGGTGTGAGAAGCTGGCC
	GAGATCATCTGGCAGAACCGGCAGCAGATCCGCAGGGCTGAGCACTTGT
	GCCAGCAGCTGCCCATCCCAGGCCCCGTGGAGGAGATGCTGGCTGAGGT
	CAACGCCACCATCACGGACATCATCTCAGCCCTGGTCACCAGCACGTTC
	ATCATCGAGAAGCAGCCTCCTCAGGTCCTGAAGACCCAGACCAAGTTTG
	CAGCCACCGTGCGCCTGCTGGTGGGGGGGAAGCTGAATGTGCACATGAA
	CCCCCCGCAGGTGAAGGCGACCATCATCAGCGAGCAGCAGGCCAAGTCC
	CTGCTCAAGAATGAGAACACCCGCAATGATTACAGCGGCGAGATCCTGA
	ACAACTGTTGCGTCATGGAGTACCACCAGGCCACTGGCACACTCAGCGC
	CCACTTCAGAAACATGTCCCTGAAACGAATCAAGAGGTCTGACCGCCGT
	GGGGCAGAGTCAGTAACGGAAGAGAAGTTCACGATCCTGTTTGACTCAC
	AGTTCAGCGTCGGTGGAAACGAGCTGGTCTTTCAAGTCAAGACCTTGTC
	GCTCCCGGTGGTGGTGATTGTTCACGGCAGCCAGGACAACAATGCCACA
	GCCACTGTCCTCTGGGACAACGCCTTTGCAGAGCCTGGCAGGGTGCCATT
	TGCCGTGCCTGACAAGGTGCTGTGGCCGCAGCTGTGTGAAGCGCTCAAC
	ATGAAATTCAAGGCTGAAGTACAGAGCAACCGGGGCTTGACCAAGGAG
	AACCTCGTGTTCCTGGCACAGAAACTGTTCAACATCAGCAGCAACCACC
	TCGAGGACTACAACAGCATGTCCGTGTCCTGGTCCCAGTTCAACCGGGA
	GAATTTGCCAGGACGGAATTACACTTTCTGGCAGTGGTTTGATGGCGTGA
	TGGAAGTATTGAAAAAACATCTCAAGCCTCACTGGAATGATGGGGCTAT
	CCTGGGTTTCGTGAACAAGCAACAGGCCCACGACCTGCTCATCAACAAG
	CCAGACGGGACCTTCCTGCTGCGCTTCAGCGACTCGGAAATCGGGGGCA
	TCACCATTGCTTGGAAGTTTGACTCTCAGGAGAGAATGTTTTGGAATCTG
	ATGCCTTTTACCACTAGAGACTTCTCTATCCGGTCCCTCGCTGACCGCCT
	GGGGGACCTGAATTACCTCATATATGTGTTTCCTGATCGGCCAAAGGATG
	AAGTATATTCTAAGTACTACACACCGGTCCCCTGTGAGCCCGCAACTGCG
	AAAGCAGCTGACGGATACGTGAAGCCACAGATCAAGCAGGTGGTCCCCG
	AGTTTGCAAATGCATCCACAGATGCTGGGAGTGGCGCCACCTACATGGA
	TCAGGCTCCTTCCCCAGTCGTGTGCCCTCAGGCTCACTACAACATGTACC
	CACCCAACCCGGACTCCGTCCTTGATACCGATGGGGACTTCGATCTGGA
	AGACACGATGGACGTGGCGCGGCGCGTGGAAGAGCTCTTAGGCCGGCCC
	ATGGACAGTCAGTGGATCCCTCACGCACAGTCATGACCAGACCTCACCA
	CCTGCAGCTTCATCGCCCTCGTGGAGGAACTTCCTGTGGATGTTTTAATT
	CCATGAATCGCTTCTCTTTGGAAACAATACTCGTAATGTGAAGTGTTAAT
	ACTAGTTGTGACTTTAGTGTCTCTGTGCATAGTGGCACTAGTGAAGGGAG
	TGCGCGTGAGTGTGAGTGCATTTGCACGTCGTGTTTTTTTCCCCGCCCCT
	GCTGTCCAGTCTAAGCCGCCACGCCAGGGCAGCGGCTGCGCTTTTTTTTA
	CCATGTGCAAAAAGGCAGTTGGTTCCCTGAACCCTGGAACCTGGCCATG
	TGTCTTCAGGGTGGCTGACCCTTGACACGTGACTATCCAAGTAAGAAAA
	GGACAGAGGAAAAAGCACCCTCTCTCTGGGGAGCCTCGGTTCCTCTGCC
	AGGTAGTCCATAGTCCAAGCAAGCATTGTCATTGTCTCCGCCTGTCTTCT
	GAGATGTAGATGACTGTCTGATGATGAAAGCCAGTACCTCCCGTGTCCC
	CTGTCCCCTTTGCATAAGGGACGGAAAGGGGAGCTGAATCAAGGGTGAT
	GGGGCAAGGGTGGTCACAGGTTTTTGGATGGGGAGTGGCTGTTTCCCGT
	TTCTGCCACTTCCGCCATCTTAACACTGGCTCCTTCCCTCTCTGCTTGCTC
	AGTCTCTATTTCTAGAACTGCCACTCAGCTTAAGTGCAAGTGTGTGTACT
	CAAGTGGAGATGTTTAACAAAATAGTGGAGAGGAAGCCAGGCCACCCA
	GCTCTGAGCGTACAGGTTCAGGTGATGCCCTGTGTTCCTTCTGTCAGGGC
	GGTGCGTTGTGCCCAAGTCCTGGCTCCAGACACTGGGCGTAGCCTGTCTG
	CGCCAGCCTCCCCAACTCTTGTCTGTGCTGTGGCCAGGCCGCGCCTGCGC
	TATCCAAGGCTTTTCTCCAAGCGTGTTGATAATGGCTTCCTGCAAACGTC
	CGGTGGTGTTTTTTGTTTCTAAATCAGGTCTTTTTTTATGTTTTTCCCATTT
	GCACCCTAATTTGACATCAAATTTCCCCCCTCCTGTATCAGGTCCTGGGT
	CCTCTGTACCCAGATCACTTCATCTCCCTTCAGTGTCACATAGTGCCCTG
	AGGATTAGGTGGTAGGAATGGGACCTGCACACGGGGCCAGCCTGCCAAG
	CAGGCAGCCAGCACTGTACAGTGCTGGGTGCCGGGTGGCCGTTGGGGAT
	TGGGGAAATGCAGTCAGTCAGCGGGTTTCCTAGGAAGCTTGGAAAACTA
	AAAGCAAAGTGAAAGCCTCAGGGTGATTTGTTCCACAGTCTCCTCTGTA
	GTGTCTCCAGAAGGAAGGAAGGGGCTGCAGTGGGCCGTCAGGGAGAGG
	GGCAAGCAGAGAGCGGTTACCACTCAGGCTTGCTGAGAGCCCTCCTTGG
	CTTCCTCTCCCAAACAAGGGCAGAACCGTGCCCAGGAGAGGAGCCCCCA
	AAACCTTATTTTTATACATGCAAGTAAATAAACATATTTTTTTTACAAAA
	ATAACTTCTGAATTTATCAGTGTTTTACTGTTAAAAGAAAATACTCCTGT
	GTAGTAAATTATTTATTGGGAGATGAGTTTTTAAAAGCTGCTGTTTGCCT
	TGCCTTGGTTTTGTACACTGATTTTTCTATGCCTGGCGGTAGCCTCTCTGC
	CTCAGGTGCTGGCCGGATGGAGGAGGTGTGAGGCCCCTCCCTGGCCCCT
	CAGAAGAAAGCTGGAACTGCCAGGGGAGTCCAGGCTTAAGGGACTCGTC
	CCCACCTGTCATGCGACTGTCCCAGTAACCCTCACGAGGGTGTGGACTCG
	ACAAATATCTAGATATATGGTGGACATGGCCCCAAGTCATGGGGAGAGT
	AGAGCAGCCTGGGCCCCCCCACCCCCAAGGTTCTAAGCTGACTTTCAAG
	TTAGGTTGGAGAAAAGGGTGCCAAAGAAGCGAGACTTCCACATAGTTTT
	TAAGCTACCCTGGATTTACTGAGGGTGTACCTGGACATGGGAGAGGTTTT
	TAACTGGAAAGTGTGTCCCCTATCTGCATGCTGGTCTCTCTCTCTCTCTGC
	CCAACTCTTGCACCCAAAAATGAGGTGAGGGCAGGTCTCCACCCACCTC
	TTGCCTGCTCACAGACCCACTCGTGAGTCGGGAAAGCCTCAGCTTTGGG
	GTGTGGGGCTTTGTAGAAGTGGAAGGAGATTTGAAGTGGCTATCTCCTA
	CAACGGAAAATATCCTTTTATAATTTTTCTTTTTAACGTTTTATTTCAGAT
	ACATATTTTAGTGTCGAGGCAGATTAGTATATAGCCACCAAAAAAGTAT
	TGTGTATAAATTGAGGCAGCCACAAAATTGTGTATTTTATGTTACAATAA
	AGGCGTCTCCTTGAAGGACAA
Modified	ATGGCCACCGGCTCCCGCACCTCCCTGCTGCTGGCCTTCGGCCTGCTGTG	8
Human GH	CCTGCCCTGGCTGCAGGAGGGCTCCGCCTTCCCCACCATCCCCCTGTCCC
	GCCTGTTCGACAACGCCATGCTGCGCGCCCACCGCCTGCACCAGCTGGC
	CTTCGACACCTACCAGGAGTTCGAGGAGGCCTACATCCCCAAGGAGCAG
	AAGTACTCCTTCCTGCAGAACCCCCAGACCTCCCTGTGCTTCTCCGAGTC
	CATCCCCACCCCCTCCAACCGCGAGGAGACCCAGCAGAAGTCCAACCTG
	GAGCTGCTGCGCATCTCCCTGCTGCTGATCCAGTCCTGGCTGGAGCCCGT
	GCAGTTCCTGCGCTCCGTGTTCGCCAACTCCCTGGTGTACGGCGCCTCCG
	ACTCCAACGTGTACGACCTGCTGAAGGACCTGGAGGAGGGCATCCAGAC
	CCTGATGGGCCGCCTGGAGGACGGCTCCCCCCGCACCGGCCAGATCTTC
	AAGCAGACCTACTCCAAGTTCGACACCAACTCCCACAACGACGACGCCC
	TGCTGAAGAACTACGGCCTGCTGTACTGCTTCCGCAAGGACATGGACAA
	GGTGGAGACCTTCCTGCGCATCGTGCAGTGCCGCTCCGTGGAGGGCTCCT
	GCGGCTTCtaa
Modified	ATGCTGCTGACCCTGATCATCCTGCTGCCCGTGGTGTCCAAGTTCTCCTT	9
Human EGF	CGTGTCCCTGTCCGCCCCCCAGCACTGGTCCTGCCCCGAGGGCACCCTG
	GCCGGCAACGGCAACTCCACCTGCGTGGGCCCCGCCCCCTTCCTGATCT
	TCTCCCACGGCAACTCCATCTTCCGCATCGACACCGAGGGCACCAACTA
	CGAGCAGCTGGTGGTGGACGCCGGCGTGTCCGTGATCATGGACTTCCAC
	TACAACGAGAAGCGCATCTACTGGGTGGACCTGGAGCGCCAGCTGCTG
	CAGCGCGTGTTCCTGAACGGCTCCCGCCAGGAGCGCGTGTGCAACATCG
	AGAAGAACGTGTCCGGCATGGCCATCAACTGGATCAACGAGGAGGTGA
	TCTGGTCCAACCAGCAGGAGGGCATCATCACCGTGACCGACATGAAGG
	GCAACAACTCCCACATCCTGCTGTCCGCCCTGAAGTACCCCGCCAACGT
	GGCCGTGGACCCCGTGGAGCGgTTCATCTTCTGGTCCTCCGAGGTGGCC
	GGCTCCCTGTACCGCGCCGACCTGGACGGCGTGGGCGTGAAGGCCCTGC
	TGGAGACCTCCGAGAAGATCACCGCCGTGTCCCTGGACGTGCTGGACA
	AGCGCCTGTTCTGGATCCAGTACAACCGCGAGGGCTCCAACTCCCTGAT
	CTGCTCCTGCGACTACGACGGCGGCTCCGTGCACATCTCCAAGCACCCC
	ACCCAGCACAACCTGTTCGCCATGTCCCTGTTCGGCGACCGCATCTTCT
	ACTCCACCTGGAAGATGAAGACCATCTGGATCGCCAACAAGCACACCG
	GCAAGGACATGGTGCGCATCAACCTGCACTCCTCCTTCGTGCCCCTGGG
	CGAGCTGAAGGTGGTGCACCCCCTGGCCCAGCCCAAGGCCGAGGACGA
	CACCTGGGAGCCCGAGCAGAAGCTGTGCAAGCTGCGCAAGGGCAACTG
	CTCCTCCACCGTGTGCGGCCAGGACCTGCAGTCCCACCTGTGCATGTGC
	GCCGAGGGCTACGCCCTGTCCCGCGACCGCAAGTACTGCGAGGACGTG
	AACGAGTGCGCCTTCTGGAACCACGGCTGCACCCTGGGCTGCAAGAAC
	ACCCCCGGCTCCTACTACTGCACCTGCCCCGTGGGCTTCGTGCTGCTGCC
	CGACGGCAAGCGgTGCCACCAGCTGGTGTCCTGCCCCCGCAACGTGTCC
	GAGTGCTCCCACGACTGCGTGCTGACCTCCGAGGGCCCCCTGTGCTTCT
	GCCCCGAGGGCTCCGTGCTGGAGCGCGACGGCAAGACCTGCTCCGGCT
	GCTCCTCCCCCGACAACGGCGGCTGCTCCCAGCTGTGCGTGCCCCTGTC
	CCCCGTGTCCTGGGAGTGCGACTGCTTCCCCGGCTACGACCTGCAGCTG
	GACGAGAAGTCCTGCGCCGCCTCCGGCCCCCAGCCCTTCCTGCTGTTCG
	CCAACTCCCAGGACATCCGCCACATGCACTTCGACGGCACCGACTACGG
	CACCCTGCTGTCCCAGCAGATGGGCATGGTGTACGCCCTGGACCACGAC
	CCCGTGGAGAACAAGATCTACTTCGCCCACACCGCCCTGAAGTGGATCG
	AGCGCGCCAACATGGACGGCTCCCAGCGCGAGCGCCTGATCGAGGAGG
	GCGTGGACGTGCCCGAGGGCCTGGCCGTGGACTGGATCGGCCGCCGCTT
	CTACTGGACCGACCGCGGCAAGTCCCTGATCGGCCGCTCCGACCTGAAC
	GGCAAGCGgTCCAAGATCATCACCAAGGAGAACATCTCCCAGCCCCGCG
	GCATCGCCGTGCACCCCATGGCCAAGCGCCTGTTCTGGACCGACACCGG
	CATCAACCCCCGCATCGAGTCCTCCTCCCTGCAGGGCCTGGGCCGCCTG
	GTGATCGCCTCCTCCGACCTGATCTGGCCCTCCGGCATCACCATCGACTT
	CCTGACCGACAAGCTGTACTGGTGCGACGCCAAGCAGTCCGTGATCGA
	GATGGCCAACCTGGACGGCTCCAAGCGCCGCCGCCTGACCCAGAACGA
	CGTGGGCCACCCCTTCGCCGTGGCCGTGTTCGAGGACTACGTGTGGTTC
	TCCGACTGGGCCATGCCCTCCGTGATCCGCGTGAACAAGCGCACCGGCA
	AGGACCGCGTGCGCCTGCAGGGCTCCATGCTGAAGCCCTCCTCCCTGGT
	GGTGGTGCACCCCCTGGCCAAGCCCGGCGCCGACCCCTGCCTGTACCAG
	AACGGCGGCTGCGAGCACATCTGCAAGAAGCGCCTGGGCACCGCCTGG
	TGCTCCTGCCGCGAGGGCTTCATGAAGGCCTCCGACGGCAAGACCTGCC
	TGGCCCTGGACGGCCACCAGCTGCTGGCCGGCGGCGAGGTGGACCTGA
	AGAACCAGGTGACCCCCCTGGACATCCTGTCCAAGACCCGCGTGTCCGA
	GGACAACATCACCGAGTCCCAGCACATGCTGGTGGCCGAGATCATGGT
	GTCCGACCAGGACGACTGCGCCCCCGTGGGCTGCTCCATGTACGCCCGC
	TGCATCTCCGAGGGCGAGGACGCCACCTGCCAGTGCCTGAAGGGCTTCG
	CCGGCGACGGCAAGCTGTGCTCCGACATCGACGAGTGCGAGATGGGCG
	TGCCCGTGTGCCCCCCCGCCTCCTCCAAGTGCATCAACACCGAGGGCGG
	CTACGTGTGCCGCTGCTCCGAGGGCTACCAGGGCGACGGCATCCACTGC
	CTGGACATCGACGAGTGCCAGCTGGGCGTGCACTCCTGCGGCGAGAAC
	GCCTCCTGCACCAACACCGAGGGCGGCTACACCTGCATGTGCGCCGGCC
	GCCTGTCCGAGCCCGGCCTGATCTGCCCCGACTCCACCCCCCCCCCCCA
	CCTGCGCGAGGACGACCACCACTACTCCGTGCGCAACTCCGACTCCGAG
	TGCCCCCTGTCCCACGACGGCTACTGCCTGCACGACGGCGTGTGCATGT
	ACATCGAGGCCCTGGACAAGTACGCCTGCAACTGCGTGGTGGGCTACAT
	CGGCGAGCGgTGCCAGTACCGCGACCTGAAGTGGTGGGAGCTGCGCCA
	CGCCGGCCACGGCCAGCAGCAGAAGGTGATCGTGGTGGCCGTGTGCGT
	GGTGGTGCTGGTGATGCTGCTGCTGCTGTCCCTGTGGGGCGCCCACTAC
	TACCGCACCCAGAAGCTGCTGTCCAAGAACCCCAAGAACCCCTACGAG
	GAGTCCTCCCGCGACGTGCGCTCCCGCCGCCCCGCCGACACCGAGGACG
	GCATGTCCTCCTGCCCCCAGCCCTGGTTCGTGGTGATCAAGGAGCACCA
	GGACCTGAAGAACGGCGGCCAGCCCGTGGCCGGCGAGGACGGCCAGGC
	CGCCGACGGCTCCATGCAGCCCACCTCCTGGCGCCAGGAGCCCCAGCTG
	TGCGGCATGGGCACCGAGCAGGGCTGCTGGATCCCCGTGTCCTCCGACA
	AGGGCTCCTGCCCCCAGGTGATGGAGCGgTCCTTCCACATGCCCTCCTA
	CGGCACCCAGACCCTGGAGGGCGGCGTGGAGAAGCCCCACTCCCTGCT
	GTCCGCCAACCCCCTGTGGCAGCAGCGCGCCCTGGACCCCCCCCACCGA
	TGGAGCTGACCCAGtaa
Modified	ATGTGGGTGACCAAGCTGCTGCCCGCCCTGCTGCTGCAGCACGTGC	10
Human HGF	TGCTGCACCTGCTGCTGCTGCCCATCGCCATCCCCTACGCCGAGGGC
	CAGCGCAAGCGCCGCAACACCATCCACGAGTTCAAGAAGTCCGCCA
	AGACCACCCTGATCAAGATCGACCCCGCCCTGAAGATCAAGACCAA
	GAAGGTGAACACCGCCGACCAGTGCGCCAACCGCTGCACCCGCAAC
	AAGGGCCTGCCCTTCACCTGCAAGGCCTTCGTGTTCGACAAGGCCC
	GCAAGCAGTGCCTGTGGTTCCCCTTCAACTCCATGTCCTCCGGCGTG
	AAGAAGGAGTTCGGCCACGAGTTCGACCTGTACGAGAACAAGGACT
	ACATCCGCAACTGCATCATCGGCAAGGGCCGCTCCTACAAGGGCAC
	CGTGTCCATCACCAAGTCCGGCATCAAGTGCCAGCCCTGGTCCTCCA
	TGATCCCCCACGAGCACTCCTTCCTGCCCTCCTCCTACCGCGGCAAG
	GACCTGCAGGAGAACTACTGCCGCAACCCCCGCGGCGAGGAGGGC
	GGCCCCTGGTGCTTCACCTCCAACCCCGAGGTGCGCTACGAGGTGT
	GCGACATCCCCCAGTGCTCCGAGGTGGAGTGCATGACCTGCAACGG
	CGAGTCCTACCGCGGCCTGATGGACCACACCGAGTCCGGCAAGATC
	TGCCAGCGgTGGGACCACCAGACCCCCCACCGCCACAAGTTCCTGC
	CCGAGCGgTACCCCGACAAGGGCTTCGACGACAACTACTGCCGCAA
	CCCCGACGGCCAGCCCCGCCCCTGGTGCTACACCCTGGACCCCCAC
	ACCCGCTGGGAGTACTGCGCCATCAAGACCTGCGCCGACAACACCA
	TGAACGACACCGACGTGCCCCTGGAGACCACCGAGTGCATCCAGGG
	CCAGGGCGAGGGCTACCGCGGCACCGTGAACACCATCTGGAACGGC
	ATCCCCTGCCAGCGgTGGGACTCCCAGTACCCCCACGAGCACGACA
	TGACCCCCGAGAACTTCAAGTGCAAGGACCTGCGCGAGAACTACTG
	CCGCAACCCCGACGGCTCCGAGTCCCCCTGGTGCTTCACCACCGAC
	CCCAACATCCGCGTGGGCTACTGCTCCCAGATCCCCAACTGCGACA
	TGTCCCACGGCCAGGACTGCTACCGCGGCAACGGCAAGAACTACAT
	GGGCAACCTGTCCCAGACCCGCTCCGGCCTGACCTGCTCCATGTGG
	GACAAGAACATGGAGGACCTGCACCGCCACATCTTCTGGGAGCCCG
	ACGCCTCCAAGCTGAACGAGAACTACTGCCGCAACCCCGACGACGA
	CGCCCACGGCCCCTGGTGCTACACCGGCAACCCCCTGATCCCCTGG
	GACTACTGCCCCATCTCCCGCTGCGAGGGCGACACCACCCCCACCA
	TCGTGAACCTGGACCACCCCGTGATCTCCTGCGCCAAGACCAAGCA
	GCTGCGCGTGGTGAACGGCATCCCCACCCGCACCAACATCGGCTGG
	ATGGTGTCCCTGCGCTACCGCAACAAGCACATCTGCGGCGGCTCCC
	TGATCAAGGAGTCCTGGGTGCTGACCGCCCGCCAGTGCTTCCCCTCC
	CGCGACCTGAAGGACTACGAGGCCTGGCTGGGCATCCACGACGTGC
	ACGGCCGCGGCGACGAGAAGTGCAAGCAGGTGCTGAACGTGTCCCA
	GCTGGTGTACGGCCCCGAGGGCTCCGACCTGGTGCTGATGAAGCTG
	GCCCGCCCCGCCGTGCTGGACGACTTCGTGTCCACCATCGACCTGCC
	CAACTACGGCTGCACCATCCCCGAGAAGACCTCCTGCTCCGTGTAC
	GGCTGGGGCTACACCGGCCTGATCAACTACGACGGCCTGCTGCGCG
	TGGCCCACCTGTACATCATGGGCAACGAGAAGTGCTCCCAGCACCA
	CCGCGGCAAGGTGACCCTGAACGAGTCCGAGATCTGCGCCGGCGCC
	GAGAAGATCGGCTCCGGCCCCTGCGAGGGCGACTACGGCGGCCCCC
	TGGTGTGCGAGCAGCACAAGATGCGCATGGTGCTGGGCGTGATCGT
	GCCCGGCCGCGGCTGCGCCATCCCCAACCGCCCCGGCATCTTCGTG
	CGCGTGGCCTACTACGCCAAGTGGATCCACAAGATCATCCTGACCT
	ACAAGGTGCCCCAGTCCtaa
Modified	ATGTCCAACCCCGGCGACGTGCGCCCCGTGCCCCACCGCTCCAAGGTGT	11
Mouse p21	GCCGCTGCCTGTTCGGCCCCGTGGACTCCGAGCAGCTGCGCCGCGACTG
	CGACGCCCTGATGGCCGGCTGCCTGCAGGAGGCCCGCGAGCGgTGGAAC
	TTCGACTTCGTGACCGAGACCCCCCTGGAGGGCAACTTCGTGTGGGAGC
	GCGTGCGCTCCCTGGGCCTGCCCAAGGTGTACCTGTCCCCCGGCTCCCGC
	TCCCGCGACGACCTGGGCGGCGACAAGCGCCCCTCCACCTCCTCCGCCCT
	GCTGCAGGGCCCCGCCCCCGAGGACCACGTGGCCCTGTCCCTGTCCTGC
	ACCCTGGTGTCCGAGCGCCCCGAGGACTCCCCCGGCGGCCCCGGCACCTC
	CCAGGGCCGCAAGCGCCGCCAGACCTCCCTGACCGACTTCTACCACTCCAA
	GCGCCGCCTGGTGTTCTGCAAGCGCAAGCCCtaa
Modified	ATGAACTTCCTGCTGTCCTGGGTGCACTGGTCCCTGGCCCTGCTGCTGTACC	12
Human	TGCACCACGCCAAGTGGTCCCAGGCCGCCCCCATGGCCGAGGGCGGCGGC
VEGF165	CAGAACCACCACGAGGTGGTGAAGTTCATGGACGTGTACCAGCGgTCCTAC
	TGCCACCCCATCGAGACCCTGGTGGACATCTTCCAGGAGTACCCCGACGAG
	ATCGAGTACATCTTCAAGCCCTCCTGCGTGCCCCTGATGCGCTGCGGCGGC
	TGCTGCAACGACGAGGGCCTGGAGTGCGTGCCCACCGAGGAGTCCAACAT
	CACCATGCAGATCATGCGCATCAAGCCCCACCAGGGCCAGCACATCGGCG
	AGATGTCCTTCCTGCAGCACAACAAGTGCGAGTGCCGCCCCAAGAAGGAC
	CGCGCCCGCCAGGAGAACCCCTGCGGCCCCTGCTCCGAGCGCCGCAAGCA
	CCTGTTCGTGCAGGACCCCCAGACCTGCAAGTGCTCCTGCAAGAACACCGA
	CTCCCGCTGCAAGGCCCGCCAGCTGGAGCTGAACGAGCGCACCTGCCGCTG
	CGACAAGCCCCGCCGCtaa
Modified	ATGGGCAAGATCTCCTCCCTGCCCACCCAGCTGTTCAAGATCTGCCT	13
Mouse IGF-1	GTGCGACTTCCTGAAGATCAAGATCCACATCATGTCCTCCTCCCACC
	TGTTCTACCTGGCCCTGTGCCTGCTGACCTTCACCTCCTCCACCACCG
	CCGGCCCCGAGACCCTGTGCGGCGCCGAGCTGGTGGACGCCCTGCA
	GTTCGTGTGCGGCCCCCGCGGCTTCTACTTCAACAAGCCCACCGGCT
	ACGGCTCCTCCATCCGCCGCGCCCCCCAGACCGGCATCGTGGACGA
	GTGCTGCTTCCGCTCCTGCGACCTGCGCCGCCTGGAGATGTACTGCG
	CCCCCCTGAAGCCCACCAAGGCCGCCCGCTCCATCCGCGCCCAGCGC
	CACACCGACATGCCCAAGACCCAGAAGGAGGTGCACCTGAAGAACA
	CCTCCCGCGGCTCCGCCGGCAACAAGACCTACCGCATGtaa
Modified	ATGCGCATGCAGCTGCTGCTGCTGATCGCCCTGTCCCTGGCCCTGGTGAC	14
Mouse IGF-1	CAACTCCGGCCCCGAGACCCTGTGCGGCGCCGAGCTGGTGGACGCCCTG
IL-2 SP	CAGTTCGTGTGCGGCCCCCGCGGCTTCTACTTCAACAAGCCCACCGGCTA
	CGGCTCCTCCATCCGCCGCGCCCCCCAGACCGGCATCGTGGACGAGTGC
	TGCTTCCGCTCCTGCGACCTGCGCCGCCTGGAGATGTACTGCGCCCCCCT
	GAAGCCCACCAAGGCCGCCCGCTCCATCCGCGCCCAGCGCCACACCGAC
	ATGCCCAAGACCCAGAAGGAGGTGCACCTGAAGAACACCTCCCGCGGCT
	CCGCCGGCAACAAGACCTACCGCATGtaa
Modified	ATGGCCACCGGCTCCCGCACCTCCCTGCTGCTGGCCTTCGGCCTGCTGTG	15
Human	CCTGCCCTGGCTGCAGGAGGGCTCCGCCATGAACTCCGACTCCGAGTGC
Secreted EGF	CCCCTGTCCCACGACGGCTACTGCCTGCACGACGGCGTGTGCATGTACAT
	CGAGGCCCTGGACAAGTACGCCTGCAACTGCGTGGTGGGCTACATCGGC
	GAGCGgTGCCAGTACCGCGACCTGAAGTGGTGGGAGCTGCGCtaa
Modified	ATGGACTACAAGGACGACGACGACAAGGGCGGCGGCGGCTCCGCCATG	16
Mouse	TGGATCCAGGCCCAGCAGCTGCAGGGCGACGCCCTGCACCAGATGCAGG
constitutively	CCCTGTACGGCCAGCACTTCCCCATCGAGGTGCGCCACTACCTGTCCCAG
activated	TGGATCGAGTCCCAGGCCTGGGACTCCATCGACCTGGACAACCCCCAGG
stat5b	AGAACATCAAGGCCACCCAGCTGCTGGAGGGCCTGGTGCAGGAGCTGCA
(stat5bCA)	GAAGAAGGCCGAGCACCAGGTGGGCGAGGACGGCTTCCTGCTGAAGAT
	CAAGCTGGGCCACTACGCCACCCAGCTGCAGTCCACCTACGACCGCTGC
	CCCATGGAGCTGGTGCGCTGCATCCGCCACATCCTGTACAACGAGCAGC
	GCCTGGTGCGCGAGGCCAACAACGGCTCCTCCCCCGCCGGCTCCCTGGC
	CGACGCCATGTCCCAGAAGCACCTGCAGATCAACCAGACCTTCGAGGAG
	CTGCGCCTGATCACCCAGGACACCGAGAACGAGCTGAAGAAGCTGCAGC
	AGACCCAGGAGTACTTCATCATCCAGTACCAGGAGTCCCTGCGCATCCA
	GGCCCAGTTCGCCCAGCTGGGCCAGCTGAACCCCCAGGAGCGCATGTCC
	CGCGAGACCGCCCTGCAGCAGAAGCAGGTGTCCCTGGAGACCTGGCTGC
	AGCGCGAGGCCCAGACCCTGCAGCAGTACCGCGTGGAGCTGGCCGAGA
	AGCACCAGAAGACCCTGCAGCTGCTGCGCAAGCAGCAGACCATCATCCT
	GGACGACGAGCTGATCCAGTGGAAGCGCCGCCAGCAGCTGGCCGGCAA
	CGGCGGCCCCCCCGAGGGCTCCCTGGACGTGCTGCAGTCCTGGTGCGAG
	AAGCTGGCCGAGATCATCTGGCAGAACCGCCAGCAGATCCGCCGCGCCG
	AGCACCTGTGCCAGCAGCTGCCCATCCCCGGCCCCGTGGAGGAGATGCT
	GGCCGAGGTGAACGCCACCATCACCGACATCATCTCCGCCCTGGTGACC
	TCCACCTTCATCATCGAGAAGCAGCCCCCCCAGGTGCTGAAGACCCAGA
	CCAAGTTCGCCGCCACCGTGCGCCTGCTGGTGGGCGGCAAGCTGAACGT
	GCACATGAACCCCCCCCAGGTGAAGGCCACCATCATCTCCGAGCAGCAG
	GCCAAGTCCCTGCTGAAGAACGAGAACACCCGCAACGACTACTCCGGCG
	AGATCCTGAACAACTGCTGCGTGATGGAGTACCACCAGGCCACCGGCAC
	CCTGTCCGCCCACTTCCGCAACATGTCCCTGAAGCGCATCAAGCGgTCCG
	ACCGCCGCGGCGCCGAGTCCGTGACCGAGGAGAAGTTCACCATCCTGTT
	CGACTCCCAGTTCTCCGTGGGCGGCAACGAGCTGGTGTTCCAGGTGAAG
	ACCCTGTCCCTGCCCGTGGTGGTGATCGTGCACGGCTCCCAGGACAACA
	ACGCCACCGCCACCGTGCTGTGGGACAACGCCTTCGCCGAGCCCGGCCG
	CGTGCCCTTCGCCGTGCCCGACAAGGTGCTGTGGCCCCAGCTGTGCGAG
	GCCCTGAACATGAAGTTCAAGGCCGAGGTGCAGTCCAACCGCGGCCTGA
	CCAAGGAGAACCTGGTGTTCCTGGCCCAGAAGCTGTTCAACATCTCCTCC
	AACCACCTGGAGGACTACAACTCCATGTCCGTGTCCTGGTCCCAGTTCAA
	CCGCGAGAACCTGCCCGGCCGCAACTACACCTTCTGGCAGTGGTTCGAC
	GGCGTGATGGAGGTGCTGAAGAAGCACCTGAAGCCCCACTGGAACGAC
	GGCGCCATCCTGGGCTTCGTGAACAAGCAGCAGGCCCACGACCTGCTGA
	TCAACAAGCCCGACGGCACCTTCCTGCTGCGCTTCTCCGACTCCGAGATC
	GGCGGCATCACCATCGCCTGGAAGTTCGACTCCCAGGAGCGCATGTTCT
	GGCACCTGATGCCCTTCACCACCCGCGACTTCTCCATCCGCTCCCTGGCC
	GACCGCCTGGGCGACCTGAACTACCTGATCTACGTGTTCCCCGACCGCCC
	CAAGGACGAGGTGTACTCCAAGTACTACACCCCCGTGCCCTGCGAGCCC
	GCCACCGCCAAGGCCGCCGACGGCTACGTGAAGCCCCAGATCAAGCAGG
	TGGTGCCCGAGTTCGCCAACGCCTCCACCGACGCCGGCTCCGGCGCCAC
	CTACATGGACCAGGCCCCCTCCCCCGTGGTGTGCCCCCAGGCCCACTACA
	ACATGTACCCCCCCAACCCCGACTCCGTGCTGGACACCGACGGCGACTT
	CGACCTGGAGGACACCATGGACGTGGCCCGCCGCGTGGAGGAGCTGCTG
	GGCCGCCCCATGGACTCCCAGTGGATCCCCCACGCCCAGTCC
Human Beta	AAGCCTCTCGGTCTGTGGCAGCAGCGTTGGCCCGGCCCCGGGAGCGGAG	17
Catenin	AGCGAGGGGAGGCGGAGACGGAGGAAGGTCTGAGGAGCAGCTTCAGTC
(CTNNB1)	CCCGCCGAGCCGCCACCGCAGGTCGAGGACGGTCGGACTCCCGCGGCGG
(variant 3)	GAGGAGCCTGTTCCCCTGAGGGTATTTGAAGTATACCATACAACTGTTTT
NCBI Ref	GAAAATCCAGCGTGGACAATGGCTACTCAAGCTGATTTGATGGAGTTGG
Seq:	ACATGGCCATGGAACCAGACAGAAAAGCGGCTGTTAGTCACTGGCAGCA
NM_001098210.2	ACAGTCTTACCTGGACTCTGGAATCCATTCTGGTGCCACTACCACAGCTC
	CTTCTCTGAGTGGTAAAGGCAATCCTGAGGAAGAGGATGTGGATACCTC
	CCAAGTCCTGTATGAGTGGGAACAGGGATTTTCTCAGTCCTTCACTCAAG
	AACAAGTAGCTGATATTGATGGACAGTATGCAATGACTCGAGCTCAGAG
	GGTACGAGCTGCTATGTTCCCTGAGACATTAGATGAGGGCATGCAGATC
	CCATCTACACAGTTTGATGCTGCTCATCCCACTAATGTCCAGCGTTTGGC
	TGAACCATCACAGATGCTGAAACATGCAGTTGTAAACTTGATTAACTATC
	AAGATGATGCAGAACTTGCCACACGTGCAATCCCTGAACTGACAAAACT
	GCTAAATGACGAGGACCAGGTGGTGGTTAATAAGGCTGCAGTTATGGTC
	CATCAGCTTTCTAAAAAGGAAGCTTCCAGACACGCTATCATGCGTTCTCC
	TCAGATGGTGTCTGCTATTGTACGTACCATGCAGAATACAAATGATGTAG
	AAACAGCTCGTTGTACCGCTGGGACCTTGCATAACCTTTCCCATCATCGT
	GAGGGCTTACTGGCCATCTTTAAGTCTGGAGGCATTCCTGCCCTGGTGAA
	AATGCTTGGTTCACCAGTGGATTCTGTGTTGTTTTATGCCATTACAACTCT
	CCACAACCTTTTATTACATCAAGAAGGAGCTAAAATGGCAGTGCGTTTA
	GCTGGTGGGCTGCAGAAAATGGTTGCCTTGCTCAACAAAACAAATGTTA
	AATTCTTGGCTATTACGACAGACTGCCTTCAAATTTTAGCTTATGGCAAC
	CAAGAAAGCAAGCTCATCATACTGGCTAGTGGTGGACCCCAAGCTTTAG
	TAAATATAATGAGGACCTATACTTACGAAAAACTACTGTGGACCACAAG
	CAGAGTGCTGAAGGTGCTATCTGTCTGCTCTAGTAATAAGCCGGCTATTG
	TAGAAGCTGGTGGAATGCAAGCTTTAGGACTTCACCTGACAGATCCAAG
	TCAACGTCTTGTTCAGAACTGTCTTTGGACTCTCAGGAATCTTTCAGATG
	CTGCAACTAAACAGGAAGGGATGGAAGGTCTCCTTGGGACTCTTGTTCA
	GCTTCTGGGTTCAGATGATATAAATGTGGTCACCTGTGCAGCTGGAATTC
	TTTCTAACCTCACTTGCAATAATTATAAGAACAAGATGATGGTCTGCCAA
	GTGGGTGGTATAGAGGCTCTTGTGCGTACTGTCCTTCGGGCTGGTGACAG
	GGAAGACATCACTGAGCCTGCCATCTGTGCTCTTCGTCATCTGACCAGCC
	GACACCAAGAAGCAGAGATGGCCCAGAATGCAGTTCGCCTTCACTATGG
	ACTACCAGTTGTGGTTAAGCTCTTACACCCACCATCCCACTGGCCTCTGA
	TAAAGGCTACTGTTGGATTGATTCGAAATCTTGCCCTTTGTCCCGCAAAT
	CATGCACCTTTGCGTGAGCAGGGTGCCATTCCACGACTAGTTCAGTTGCT
	TGTTCGTGCACATCAGGATACCCAGCGCCGTACGTCCATGGGTGGGACA
	CAGCAGCAATTTGTGGAGGGGGTCCGCATGGAAGAAATAGTTGAAGGTT
	GTACCGGAGCCCTTCACATCCTAGCTCGGGATGTTCACAACCGAATTGTT
	ATCAGAGGACTAAATACCATTCCATTGTTTGTGCAGCTGCTTTATTCTCC
	CATTGAAAACATCCAAAGAGTAGCTGCAGGGGTCCTCTGTGAACTTGCT
	CAGGACAAGGAAGCTGCAGAAGCTATTGAAGCTGAGGGAGCCACAGCT
	CCTCTGACAGAGTTACTTCACTCTAGGAATGAAGGTGTGGCGACATATG
	CAGCTGCTGTTTTGTTCCGAATGTCTGAGGACAAGCCACAAGATTACAA
	GAAACGGCTTTCAGTTGAGCTGACCAGCTCTCTCTTCAGAACAGAGCCA
	ATGGCTTGGAATGAGACTGCTGATCTTGGACTTGATATTGGTGCCCAGGG
	AGAACCCCTTGGATATCGCCAGGATGATCCTAGCTATCGTTCTTTTCACT
	CTGGTGGATATGGCCAGGATGCCTTGGGTATGGACCCCATGATGGAACA
	TGAGATGGGTGGCCACCACCCTGGTGCTGACTATCCAGTTGATGGGCTG
	CCAGATCTGGGGCATGCCCAGGACCTCATGGATGGGCTGCCTCCAGGTG
	ACAGCAATCAGCTGGCCTGGTTTGATACTGACCTGTAAATCATCCTTTAG
	GAGTAACAATACAAATGGATTTTGGGAGTGACTCAAGAAGTGAAGAATG
	CACAAGAATGGATCACAAGATGGAATTTATCAAACCCTAGCCTTGCTTG
	TTAAATTTTTTTTTTTTTTTTTTTAAGAATATCTGTAATGGTACTGACTTTG
	CTTGCTTTGAAGTAGCTCTTTTTTTTTTTTTTTTTTTTTTTTTGCAGTAACT
	GTTTTTTAAGTCTCTCGTAGTGTTAAGTTATAGTGAATACTGCTACAGCA
	ATTTCTAATTTTTAAGAATTGAGTAATGGTGTAGAACACTAATTCATAAT
	CACTCTAATTAATTGTAATCTGAATAAAGTGTAACAATTGTGTAGCCTTT
	TTGTATAAAATAGACAAATAGAAAATGGTCCAATTAGTTTCCTTTTTAAT
	ATGCTTAAAATAAGCAGGTGGATCTATTTCATGTTTTTGATCAAAAACTA
	TTTGGGATATGTATGGGTAGGGTAAATCAGTAAGAGGTGTTATTTGGAA
	CCTTGTTTTGGACAGTTTACCAGTTGCCTTTTATCCCAAAGTTGTTGTAAC
	CTGCTGTGATACGATGCTTCAAGAGAAAATGCGGTTATAAAAAATGGTT
	CAGAATTAAACTTTTAATTCATTC
Modified	ATGCGCATGCAGCTGCTGCTGCTGATCGCCCTGTCCCTGGCCCTGGTGAC	18
Mouse IGF-1	CAACTCC
IL-2 SP
Inserted
Opening
Sequence
Modified	ATGGCCACCGGCTCCCGCACCTCCCTGCTGCTGGCCTTCGGCCTGCTGTG	19
Human EGF	CCTGCCCTGGCTGCAGGAGGGCTCCGCCATGAACTCC
Secreted
Inserted
Opening
Sequence
Modified	ATGGACTACAAGGACGACGACGACAAGGGCGGCGGCGGCTCC	20
Mouse
stat5bCA
Inserted
Opening
Sequence
Modified	GCCACCCAGGCCGACCTGATGGAGCTGGACATGGCCATGGAGCCCGACC	21
Human β	GCAAGGCCGCCGTGTCCCACTGGCAGCAGCAGTCCTACCTGGACTCCGG
catenin	CATCCACTCCGGCGCCACCACCACCGCCCCCTACCTGTCCGGCAAGGGC
(activated)	AACCCCGAGGAGGAGGACGTGGACACCTCCCAGGTGCTGTACGAGTGGG
	AGCAGGGCTTCTCCCAGTCCTTCACCCAGGAGCAGGTGGCCGACATCGA
	CGGCCAGTACGCCATGACCCGCGCCCAGCGCGTGCGCGCCGCCATGTTC
	CCCGAGACCCTGGACGAGGGCATGCAGATCCCCTCCACCCAGTTCGACG
	CCGCCCACCCCACCAACGTGCAGCGCCTGGCCGAGCCCTCCCAGATGCT
	GAAGCACGCCGTGGTGAACCTGATCAACTACCAGGACGACGCCGAGCTG
	GCCACCCGCGCCATCCCCGAGCTGACCAAGCTGCTGAACGACGAGGACC
	AGGTGGTGGTGAACAAGGCCGCCGTGATGGTGCACCAGCTGTCCAAGAA
	GGAGGCCTCCCGCCACGCCATCATGCGCTCCCCCCAGATGGTGTCCGCC
	ATCGTGCGCACCATGCAGAACACCAACGACGTGGAGACCGCCCGCTGCA
	CCGCCGGCACCCTGCACAACCTGTCCCACCACCGCGAGGGCCTGCTGGC
	CATCTTCAAGTCCGGCGGCATCCCCGCCCTGGTGAAGATGCTGGGCTCCC
	CCGTGGACTCCGTGCTGTTCTACGCCATCACCACCCTGCACAACCTGCTG
	CTGCACCAGGAGGGCGCCAAGATGGCCGTGCGCCTGGCCGGCGGCCTGC
	AGAAGATGGTGGCCCTGCTGAACAAGACCAACGTGAAGTTCCTGGCCAT
	CACCACCGACTGCCTGCAGATCCTGGCCTACGGCAACCAGGAGTCCAAG
	CTGATCATCCTGGCCTCCGGCGGCCCCCAGGCCCTGGTGAACATCATGCG
	CACCTACACCTACGAGAAGCTGCTGTGGACCACCTCCCGCGTGCTGAAG
	GTGCTGTCCGTGTGCTCCTCCAACAAGCCCGCCATCGTGGAGGCCGGCG
	GCATGCAGGCCCTGGGCCTGCACCTGACCGACCCCTCCCAGCGCCTGGT
	GCAGAACTGCCTGTGGACCCTGCGCAACCTGTCCGACGCCGCCACCAAG
	CAGGAGGGCATGGAGGGCCTGCTGGGCACCCTGGTGCAGCTGCTGGGCT
	CCGACGACATCAACGTGGTGACCTGCGCCGCCGGCATCCTGTCCAACCT
	GACCTGCAACAACTACAAGAACAAGATGATGGTGTGCCAGGTGGGCGGC
	ATCGAGGCCCTGGTGCGCACCGTGCTGCGCGCCGGCGACCGCGAGGACA
	TCACCGAGCCCGCCATCTGCGCCCTGCGCCACCTGACCTCCCGCCACCAG
	GAGGCCGAGATGGCCCAGAACGCCGTGCGCCTGCACTACGGCCTGCCCG
	TGGTGGTGAAGCTGCTGCACCCCCCCTCCCACTGGCCCCTGATCAAGGCC
	ACCGTGGGCCTGATCCGCAACCTGGCCCTGTGCCCCGCCAACCACGCCC
	CCCTGCGCGAGCAGGGCGCCATCCCCCGCCTGGTGCAGCTGCTGGTGCG
	CGCCCACCAGGACACCCAGCGCCGCACCTCCATGGGCGGCACCCAGCAG
	CAGTTCGTGGAGGGCGTGCGCATGGAGGAGATCGTGGAGGGCTGCACCG
	GCGCCCTGCACATCCTGGCCCGCGACGTGCACAACCGCATCGTGATCCG
	CGGCCTGAACACCATCCCCCTGTTCGTGCAGCTGCTGTACTCCCCCATCG
	AGAACATCCAGCGCGTGGCCGCCGGCGTGCTGTGCGAGCTGGCCCAGGA
	CAAGGAGGCCGCCGAGGCCATCGAGGCCGAGGGCGCCACCGCCCCCCTG
	ACCGAGCTGCTGCACTCCCGCAACGAGGGCGTGGCCACCTACGCCGCCG
	CCGTGCTGTTCCGCATGTCCGAGGACAAGCCCCAGGACTACAAGAAGCG
	CCTGTCCGTGGAGCTGACCTCCTCCCTGTTCCGCACCGAGCCCATGGCCT
	GGAACGAGACCGCCGACCTGGGCCTGGACATCGGCGCCCAGGGCGAGC
	CCCTGGGCTACCGCCAGGACGACCCCTCCTACCGCTCCTTCCACTCCGGC
	GGCTACGGCCAGGACGCCCTGGGCATGGACCCCATGATGGAGCACGAGA
	TGGGCGGCCACCACCCCGGCGCCGACTACCCCGTGGACGGCCTGCCCGA
	CCTGGGCCACGCCCAGGACCTGATGGACGGCCTGCCCCCCGGCGACTCC
	AACCAGCTGGCCTGGTTCGACACCGACCTG
Human yes-	CTCAGTCGGGCGCAGCCGCCGCCAGGGAAAAGAAAGGGAGGAAGGAAG	22
associated	GAACAAGAAAAGGAAATAAAGAGAAAGGGGAGGCGGGGAAAGGCAAC
protein 1	GAGCTGTCCGGCCTCCGTCAAGGGAGTTGGAGGGAAAAAGTTCTCAGGC
(YAP-1)	GCCGCAGGTCCGAGTGCCTCGCAGCCCCTCCCGAGGCGCAGCCGCCAGA
(variant 3)	CCAGTGGAGCCGGGGCGCAGGGCGGGGGCGGAGGCGCCGGGGCGGGGG
NCBI Ref	ATGCGGGGCCGCGGCGCAGCCCCCCGGCCCTGAGAGCGAGGACAGCGC
Seq:	CGCCCGGCCCGCAGCCGTCGCCGCTTCTCCACCTCGGCCCGTGGAGCCG
NM_001195044.2	GGGCGTCCGGGCGTAGCCCTCGCTCGCCTGGGTCAGGGGGTGCGCGTCG
	GGGGAGGCAGAAGCCATGGATCCCGGGCAGCAGCCGCCGCCTCAACCG
	GCCCCCCAGGGCCAAGGGCAGCCGCCTTCGCAGCCCCCGCAGGGGCAGG
	GCCCGCCGTCCGGACCCGGGCAACCGGCACCCGCGGCGACCCAGGCGGC
	GCCGCAGGCACCCCCCGCCGGGCATCAGATCGTGCACGTCCGCGGGGAC
	TCGGAGACCGACCTGGAGGCGCTCTTCAACGCCGTCATGAACCCCAAGA
	CGGCCAACGTGCCCCAGACCGTGCCCATGAGGCTCCGGAAGCTGCCCGA
	CTCCTTCTTCAAGCCGCCGGAGCCCAAATCCCACTCCCGACAGGCCAGTA
	CTGATGCAGGCACTGCAGGAGCCCTGACTCCACAGCATGTTCGAGCTCA
	TTCCTCTCCAGCTTCTCTGCAGTTGGGAGCTGTTTCTCCTGGGACACTGA
	CCCCCACTGGAGTAGTCTCTGGCCCAGCAGCTACACCCACAGCTCAGCA
	TCTTCGACAGTCTTCTTTTGAGATACCTGATGATGTACCTCTGCCAGCAG
	GTTGGGAGATGGCAAAGACATCTTCTGGTCAGAGATACTTCTTAAATCA
	CATCGATCAGACAACAACATGGCAGGACCCCAGGAAGGCCATGCTGTCC
	CAGATGAACGTCACAGCCCCCACCAGTCCACCAGTGCAGCAGAATATGA
	TGAACTCGGCTTCAGGTCCTCTTCCTGATGGATGGGAACAAGCCATGACT
	CAGGATGGAGAAATTTACTATATAAACCATAAGAACAAGACCACCTCTT
	GGCTAGACCCAAGGCTTGACCCTCGTTTTGCCATGAACCAGAGAATCAG
	TCAGAGTGCTCCAGTGAAACAGCCACCACCCCTGGCTCCCCAGAGCCCA
	CAGGGAGGCGTCATGGGTGGCAGCAACTCCAACCAGCAGCAACAGATG
	CGACTGCAGCAACTGCAGATGGAGAAGGAGAGGCTGCGGCTGAAACAG
	CAAGAACTGCTTCGGCAGGAGTTAGCCCTGCGTAGCCAGTTACCAACAC
	TGGAGCAGGATGGTGGGACTCAAAATCCAGTGTCTTCTCCCGGGATGTC
	TCAGGAATTGAGAACAATGACGACCAATAGCTCAGATCCTTTCCTTAAC
	AGTGGCACCTATCACTCTCGAGATGAGAGTACAGACAGTGGACTAAGCA
	TGAGCAGCTACAGTGTCCCTCGAACCCCAGATGACTTCCTGAACAGTGT
	GGATGAGATGGATACAGGTGATACTATCAACCAAAGCACCCTGCCCTCA
	CAGCAGAACCGTTTCCCAGACTACCTTGAAGCCATTCCTGGGACAAATG
	TGGACCTTGGAACACTGGAAGGAGATGGAATGAACATAGAAGGAGAGG
	AGCTGATGCCAAGTCTGCAGGAAGCTTTGAGTTCTGACATCCTTAATGAC
	ATGGAGTCTGTTTTGGCTGCCACCAAGCTAGATAAAGAAAGCTTTCTTAC
	ATGGTTATAGAGCCCTCAGGCAGACTGAATTCTAAATCTGTGAAGGATC
	TAAGGAGACACATGCACCGGAAATTTCCATAAGCCAGTTGCAGTTTTCA
	GGCTAATACAGAAAAAGATGAACAAACGTCCAGCAAGATACTTTAATCC
	TCTATTTTGCTCTTCCTTGTCCATTGCTGCTGTTAATGTATTGCTGACCTC
	TTTCACAGTTGGCTCTAAAGAATCAAAAGAAAAAAACTTTTTATTTCTTT
	TGCTATTAAAACTACTGTTCATTTTGGGGGCTGGGGGAAGTGAGCCTGTT
	TGGATGATGGATGCCATTCCTTTTGCCCAGTTAAATGTTCACCAATCATT
	TTAACTAAATACTCAGACTTAGAAGTCAGATGCTTCATGTCACAGCATTT
	AGTTTGTTCAACAGTTGTTTCTTCAGCTTCCTTTGTCCAGTGGAAAAACA
	TGATTTACTGGTCTGACAAGCCAAAAATGTTATATCTGATATTAAATACT
	TAATGCTGATTTGAAGAGATAGCTGAAACCAAGGCTGAAGACTGTTTTA
	CTTTCAGTATTTTCTTTTCCTCCTAGTGCTATCATTAGTCACATAATGACC
	TTGATTTTATTTTAGGAGCTTATAAGGCATGAGACAATTTCCATATAAAT
	ATATTAATTATTGCCACATACTCTAATATAGATTTTGGTGGATAATTTTGT
	GGGTGTGCATTTTGTTCTGTTTTGTTGGGTTTTTTGTTTTTTTTGTTTTTGG
	CAGGGTCGGTGGGGGGGTTGGTTGGTTGGTTGGTTTTGTCGGAACCTAG
	GCAAATGACCATATTAGTGAATCTGTTAATAGTTGTAGCTTGGGATGGTT
	ATTGTAGTTGTTTTGGTAAAATCTTCATTTCCTGGTTTTTTTTACCACCTT
	ATTTAAATCTCGATTATCTGCTCTCTCTTTTATATACATACACACACCCAA
	ACATAACATTTATAATAGTGTGGTAGTGGAATGTATCCTTTTTTAGGTTT
	CCCTGCTTTCCAGTTAATTTTTAAAATGGTAGCGCTTTGTATGCATTTAGA
	ATACATGACTAGTAGTTTATATTTCACTGGTAGTTTAAATCTGGTTGGGG
	CAGTCTGCAGATGTTTGAAGTAGTTTAGTGTTCTAGAAAGAGCTATTACT
	GTGGATAGTGCCTAGGGGAGTGCTCCACGCCCTCTGGGCATACGGTAGA
	TATTATCTGATGAATTGGAAAGGAGCAAACCAGAAATGGCTTTATTTTCT
	CCCTTGGACTAATTTTTAAGTCTCGATTGGAATTCAGTGAGTAGGTTCAT
	AATGTGCATGACAGAAATAAGCTTTATAGTGGTTTACCTTCATTTAGCTT
	TGGAAGTTTTCTTTGCCTTAGTTTTGGAAGTAAATTCTAGTTTGTAGTTCT
	CATTTGTAATGAACACATTAACGACTAGATTAAAATATTGCCTTCAAGAT
	TGTTCTTACTTACAAGACTTGCTCCTACTTCTATGCTGAAAATTGACCCTG
	GATAGAATACTATAAGGTTTTGAGTTAGCTGGAAAAGTGATCAGATTAA
	TAAATGTATATTGGTAGTTGAATTTAGCAAAGAAATAGAGATAATCATG
	ATTATACCTTTATTTTTACAGGAAGAGATGATGTAACTAGAGTATGTGTC
	TACAGGAGTAATAATGGTTTCCAAAGAGTATTTTTTAAAGGAACAAAAC
	GAGCATGAATTAACTCTTCAATATAAGCTATGAAGTAATAGTTGGTTGTG
	AATTAAAGTGGCACCAGCTAGCACCTCTGTGTTTTAAGGGTCTTTCAATG
	TTTCTAGAATAAGCCCTTATTTTCAAGGGTTCATAACAGGCATAAAATCT
	CTTCTCCTGGCAAAAGCTGCTATGAAAAGCCTCAGCTTGGGAAGATAGA
	TTTTTTTCCCCCCAATTACAAAATCTAAGTATTTTGGCCCTTCAATTTGGA
	GGAGGGCAAAAGTTGGAAGTAAGAAGTTTTATTTTAAGTACTTTCAGTG
	CTCAAAAAAATGCAATCACTGTGTTGTATATAATAGTTCATAGGTTGATC
	ACTCATAATAATTGACTCTAAGGCTTTTATTAAGAAAACAGCAGAAAGA
	TTAAATCTTGAATTAAGTCTGGGGGGAAATGGCCACTGCAGATGGAGTT
	TTAGAGTAGTAATGAAATTCTACCTAGAATGCAAAATTGGGTATATGAA
	TTACATAGCATGTTGTTGGGATTTTTTTTAATGTGCAGAAGATCAAAGCT
	ACTTGGAAGGAGTGCCTATAATTTGCCAGTAGCCACAGATTAAGATTAT
	ATCTTATATATCAGCAGATTAGCTTTAGCTTAGGGGGAGGGTGGGAAAG
	TTTGGGGGGGGGGTTGTGAAGATTTAGGGGGACCTTGATAGAGAACTTT
	ATAAACTTCTTTCTCTTTAATAAAGACTTGTCTTACACCGTGCTGCCATTA
	AAGGCAGCTGTTCTAGAGTTTCAGTCACCTAAGTACACCCACAAAACAA
	TATGAATATGGAGATCTTCCTTTACCCCTCAACTTTAATTTGCCCAGTTAT
	ACCTCAGTGTTGTAGCAGTACTGTGATACCTGGCACAGTGCTTTGATCTT
	ACGATGCCCTCTGTACTGACCTGAAGGAGACCTAAGAGTCCTTTCCCTTT
	TTGAGTTTGAATCATAGCCTTGATGTGGTCTCTTGTTTTATGTCCTTGTTC
	CTAATGTAAAAGTGCTTAACTGCTTCTTGGTTGTATTGGGTAGCATTGGG
	ATAAGATTTTAACTGGGTATTCTTGAATTGCTTTTACAATAAACCAATTT
	TATAATCTTTAAATTTATCAACTTTTTACATTTGTGTTATTTTCAGTCAGG
	GCTTCTTAGATCTACTTATGGTTGATGGAGCACATTGATTTGGAGTTTCA
	GATCTTCCAAAGCACTATTTGTTGTAATAACTTTTCTAAATGTAGTGCCTT
	TAAAGGAAAAATGAACACAGGGAAGTGACTTTGCTACAAATAATGTTGC
	TGTGTTAAGTATTCATATTAAATACATGCCTTCTATATGGAACATGGCAG
	AAAGACTGAAAAATAACAGTAATTAATTGTGTAATTCAGAATTCATACC
	AATCAGTGTTGAAACTCAAACATTGCAAAAGTGGGTGGCAATATTCAGT
	GCTTAACACTTTTCTAGCGTTGGTACATCTGAGAAATGAGTGCTCAGGTG
	GATTTTATCCTCGCAAGCATGTTGTTATAAGAATTGTGGGTGTGCCTATC
	ATAACAATTGTTTTCTGTATCTTGAAAAAGTATTCTCCACATTTTAAATGT
	TTTATATTAGAGAATTCTTTAATGCACACTTGTCAAATATATATATATAG
	TACCAATGTTACCTTTTTATTTTTTGTTTTAGATGTAAGAGCATGCTCATA
	TGTTAGGTACTTACATAAATTGTTACATTATTTTTTCTTATGTAATACCTT
	TTTGTTTGTTTATGTGGTTCAAATATATTCTTTCCTTAAACTCTTC
Human	AGTCTCACCACTAGCCGCAGACGCGAGCGGCGGGGGGGCGGGGGCGCA	23
wingless-type	GAGGCGCCGGCAGCCGTGACGAGGCGCTCCCGGAGCTGAGCGCTTCTGC
MMTV	TCCGGGCACGCATGGCGCCCGCACACGGAGTCTGACCTGATGTAGACGC
integration	AAGGGGGTTAATATGAACGTCCCTCTCGGTGGAATCTGGCTCTGGCTCCC
site family,	TCTGCTCTTGACCTGGCTCACCCCTGAGGTCAGCTCTTCATGGTGGTACA
member 2	TGAGAGCTACAGGTGGCTCCTCCAGGGTGATGTGTGACAATGTGCCAGG
(WNT2)	CCTGGTGAGCCGGCAGCGTCAGCTGTGCCACCGACACCCAGATGTGATG
NCBI Ref	CGTGCCATTGGCCTGGGTGTGGCTGAGTGGACTGCAGAGTGCCAACACC
Seq:	AGTTCCGCCAGCATCGCTGGAACTGCAACACCCTGGACAGAGATCACAG
NM_023653.5	CCTCTTTGGCCGGGTCCTCCTCCGAAGTAGTCGGGAATCGGCCTTTGTTT
	ACGCCATCTCTTCAGCTGGCGTTGTATTTGCCATCACCAGGGCCTGTAGC
	CAAGGAGAATTAAAGTCCTGCTCCTGTGATCCAAAGAAGAAAGGAAGTG
	CCAAGGACAGCAAAGGCACCTTCGACTGGGGTGGCTGCAGTGACAATAT
	TGACTACGGGATCAAGTTTGCCCGTGCCTTTGTAGATGCCAAGGAGAGG
	AAAGGCAAGGATGCCAGAGCCCTGATGAACCTTCACAACAACAGAGCTG
	GAAGGAAGGCTGTAAAGCGCTTCTTGAAACAAGAATGCAAGTGTCATGG
	TGTGAGTGGCTCCTGTACTCTGAGGACATGCTGGCTGGCCATGGCTGACT
	TCAGGAAAACAGGCGACTATCTCTGGAGGAAGTACAATGGGGCCATCCA
	GGTAGTCATGAACCAGGATGGCACTGGCTTCACTGTAGCCAATAAGAGG
	TTTAAGAAGCCAACGAAAAATGACCTCGTGTATTTTGAGAATTCTCCAG
	ACTACTGTATCAGGGACCGAGAGGCAGGCTCCCTGGGTACAGCGGGCCG
	TGTGTGCAACTTGACTTCCCGAGGCATGGACAGCTGCGAAGTTATGTGTT
	GTGGGAGAGGCTATGACACATCCCACGTCACCCGGATGACCAAGTGTGA
	GTGTAAATTCCACTGGTGCTGTGCCGTGCGCTGTCAGGACTGCCTGGAGG
	CCCTGGACGTGCACACATGCAAGGCCCCCAAGAGTGCCGACTGGGCGAC
	GCCTACATGACCTCAGCAGAGGTCATATTCGCCTTTTCTTCCCTCAAGGA
	CTCCAATTACATCTTCAAGGACACTGGACCTCTGGGTTGTTTTCAGGGGC
	TCTTTCTTAAGGCATGAAGCCTTCATCTCAAGAGAAACCCCCTTTCCCCT
	CTCTGGGGGCCCCAGGACTGGGAACCACCTGCTGCACATAAGTACACCC
	TATTCTGTCTATCTTGGGCATTCTGATGTCACCTCTCTTCCTGCTGATTTC
	TTTTTGGAAATGGCATGACAGGCTGTTAGAGGAGGAGGGTCATAGCCCC
	CCACCACTGTCACCTAGACATTTCCTCTTTGGCTGCGGGGAGAAACATCA
	CATAGCGAAGGAACTTCCTCTGTGTTTTCCCAGATTCCAACAACCCAGAA
	AGTCTGTGTTTCCCTGGGGCGCGGGGTAGGGATGGAAAGCAGAATGAGC
	TGACACCAAAATTTCCTCGGATTTTTTTAAAAAAAGAGTAAGCAAGGGC
	TTTAACTAAGTGATAGCTGTTGATAGCATCCTTGGTGACTTTCTAGAGAA
	AGATGGCTTCCAATAAACATCAGGTTAAAACATGTATGTCTTCAAAGAA
	TTTATTGGATATTTATTGGCTATTGGATATAATAGGGTGAGAATGTTTGT
	CCTTTCAGACTGTGTTATTTTTGAACTTTCCTGTCAGCCAACACCTTAGAA
	AGTGATTGCTATTCCTCACTGTCCCATCAGTTTAAGGATTCTTAAGAGAT
	GAGACTTCTCAGTGTGCTCTGGAGAGAATCTGAAAGGGGAATGGATGAT
	CTAGCAATATTATTTAACTACTGGGTAAATATGGTTTAAAAATAATAATA
	ACTTTGTGAGTGGAATATCATAAATGTGCTTGTATGGC
Human	ACGAGCGCCTAGTGGCGCGAGGAGATGCGAGAGTGCACCGGCCGCCTGC	24
wingless-type	ACCATGCGCCCCGCGCCCGCGCTGGCCCTGGCTGCGCTCTGCCTGCTGGT
MMTV	GCTGCCTGCCGCTGCCGCCGCCGCCGCCTACTTCGGCCTGACCGGTCGTG
integration	AGGTCCTGACACCCTTCCCAGGCCTGGGTACGGCAGCAGCCCCGGCACA
site family,	GGCTGGTGCTCACCTGAAGCAGTGTGACCTACTGAAGCTGTCCAGGCGG
member 9B	CAGAAGCAGCTCTGCAGGCGGGAGCCCGGCCTGGCTGAGACCCTGAGGG
(WNT9B)	ATGCTGCACACCTGGGGCTGCTGGAATGTCAGTTCCAGTTCAGGCAGGA
NCBI Ref	GCGCTGGAACTGCAGCCTGGAGGGGAGGACTGGCCTGCTCCAGAGAGGC
Seq:	TTTAAGGAGACGGCCTTCCTGTATGCAGTGTCTGCAGCTGCCCTCACGCA
NM_011719.4	TGCACTGGCCAGGGCCTGCAGTGCTGGGCGCATGGAGCGCTGTACTTGT
	GACGACTCCCCAGGCCTGGAGAGCCGGCAGGCCTGGCAGTGGGGTGTGT
	GTGGTGACAATCTGAAGTACAGCACCAAGTTCCTCAGCAACTTCCTGGG
	GCCCAAGAGAGGAAGCAAGGACCTGAGGGCGAGGGCTGACGCCCACAA
	CACCCACGTGGGCATCAAGGCTGTGAAGAGCGGCCTGAGAACAACCTGC
	AAGTGCCATGGTGTGTCAGGCTCCTGTGCTGTTCGTACCTGTTGGAAGCA
	GCTCTCCCCGTTTCGCGAGACCGGCCAGGTGCTGAAGCTACGCTATGAC
	ACGGCTGTCAAGGTGTCCAGTGCCACCAACGAGGCCTTGGGTCGTCTGG
	AGCTATGGGCCCCCGCTAAGCCAGGTGGTCCCGCCAAGGGCCTAGCCCC
	TCGTCCCGGGGACCTGGTCTACATGGAAGATTCTCCCAGCTTCTGCCGGC
	CCAGCAAGTACTCTCCGGGCACGGCAGGCAGGGTGTGTTCTCGAGACTC
	CAGTTGCAGCAGCCTATGCTGTGGGCGAGGCTACGACACCCAGAGCCGC
	ATGGTGGTTTTCTCCTGCCACTGTCAGGTGCAGTGGTGCTGCTACGTGGA
	GTGCCAGCAGTGTGCACAGCAGGAGCTCGTGTATACCTGCAAGCGCTAG
	GCCTCCACAGCGAATCCCGCGGAACAGCGCGCAAGCGCGCACCTGTCGA
	CGCACCTGCCGTGCACAAGAGTGTGCGACTCATCTCTCTTCCCCAACAGA
	TGGTTGGCCAGCCCTTCTGCCTTCCCCGACACTCAGCAAAGAGAAAGAA
	AGCCCTGCCTCCTAGTCCCAGGATCACCAACCTGCTGGAGGACTTGGGG
	CCGGAGAACAGACTGAGAAGGGGAATCTTTGAGGACCAGGGTAGGGCA
	GGAATGATGCTGTGCGGGAAGAGAGAAACATCCTCCTATCTCAAGGCCA
	AAAACTGGGAGGATGGGGAAGAGGGAGGCGGAGCCAGCTGGAGTGTGG
	GGTCAGGGCATCCATCTGGGCGTGGCCGATCTCTTGTGGTCCCACTCTAA
	TAGCAGAGCGCTCTGGGTGCTGCATGCCTACCCTGCTCTTGTGGCTTCGT
	GCACTGGAGACTTCGAAATGTTTATTAGGAGCAAGGGAAGCACTTTAGG
	CTTGGGTGGATTGAGTCGCAGAGCCCATGCCCTGAAGTCTTACGTCCTGG
	CACTCAGGGCTGCCACCTTGTCTCCTTGTCTTGAGATCCCCTGTCCCCCA
	AAGCCATTGAGCTCTGCTCAACGAGACCCCTAATATGTATAAGAAGGGT
	GCAGGAGCCAGTCTCCTCGGTGAGACTCAGATAAACATAACTAGGGTTG
	AGCGGGGAGACAGTGACCCTTTCTCTTTCCTTTGGTCCAAGGAACCTTTA
	ATCACAGCCCAGAGGTGGAGAGAGGCAGGGTCCAAATGCCTGGAAGAG
	ATATGACAGGCTCTGTATTGAGATACCACTCTGGAGTGTGTCCTACCAAT
	TCCTGTGACCAGGGACCCCCAAGAACCGAGGGGCCCCCATCCATGTTAG
	TGATACATAAGAACGAGTGACTCATGGGCCACACGTCTGCTTCCACCCC
	CTGCTCTCAAAGATGCTTGTGCAGGCTTTTTTGCCATTGCTAAGTCTTTGC
	CAAGTCTGCCTCCTCAATGGTCTTACTCATTTACTAACGACCTGTCACTT
	GGGCTCCCACCAGAGGAACAAAATGACTGCTGGTGAATCCTTTGGTCAT
	TTTTAATGCCCCCATCAAGGCCCTCTGTGAGAGGAGAGGAAGTAGTGTA
	CAGGTACAGGCTCACACGTGCACACACTCAGCCTAGCCAGGCACAGACA
	TCCCAAGGAGCAGTGCGGCGTCTCTCCAGCCCAGGGCAAAGACCTCACT
	GGGGTCACTTCTGGAGGCTGTGAGCTACTCCAGGGCAGGGCCCAAGGCC
	AACCAGGAGGAAGTGACCTCCTTTGGGAAGCCTTTGGCCATGTGGCTGG
	CTGTGCTGCACCCTCCTGTGAGCTTCCTTCCACCCTGAAATCTGTTGGGG
	TTACTGTCTCTCTAAGGGAGCAGGAAGCTTCGGAATCAGCCGGTACTCA
	GCACTACTGGCCCTGCCAGCTCCAGGAAAGAGACACTGTGGCGGAGAGG
	TCCGTGGGGCAGAAGGGGCTACCCTTTCTTCAGTGCCTCCGGGCAGCAT
	GCTGGGAAGATCTTTGATGGTGGAAAGCCCCGAGGCGGAGCCACCGTGA
	CCTGAGACCCTTCTCTGGGACGACTTTGCCACCCACCCGCAGCTTGGCAG
	GAGGGGTAAACAGATTGGGAGCTGCTTTCTACTTCCCTGATGAAGACAG
	ATGTGTTCCTTGGCAACCCAAGGCATCCTTCTCTATGACCCTAATCCTGC
	TCTGGCTCGAGGGTACAAGGCAAGAATGGAGCCTGGCAAAACTTGGGGA
	CTAGAACACCTGGACCTACAGCCAAATCACCTGTACCCTGACTCTATGGC
	CAGGAGGGCCAGGGGTGGAGGAGGGTTAAAGATGAACTTGAAGTTGAG
	GCTGAGGCTGACCAACCATTAAGACTGGTGCCTTAAGGCACCCTCAGTC
	AGGTCCTCTCCCTCCCTTCTCCATTCTTTCTCCAAGGCCCCGTTCCCCCTA
	AAATCCCACCATAGCCATGCTGGGTCCCCCCTTCCCCCACACTGGAACTT
	TAAGGAAGATATTCACAGGGTATTTCTGCCTACCTCATACATGTAATTTT
	CAAAAAAAATTAATTTATATAGTTAAGATATATGGGAAAGTATTTATGTT
	ATTTATATATCTTCTCTATTTCCTGGGCACCATATGGGGGGTTGTGTGTTT
	ACCCAGAAGCCTCTGAGGAAACATGGCTGGGTCTGTCTGGGGCCTCGCA
	GAGCTGGATGCGCATAGCTGAGAGGTCACAGCTCCTGTGTCTCACTGTCT
	TGGAGCTCGGGAAGCACATGTACCTCCTGAGATAAACCCCGTGACACCA
	AGCAGGGCCTTCCTTGTGAAGTCTGTGGATTCTCTGCCTCTGGCCCCAGA
	GGCCTTTCTGCTCTGGCCCAAGGGTTTTGCTCATAAAGGACAAAAAGGG
	TGAGCAGCTCTGGATTTGTAAAGCACTTTCCATCTTCAGAAACACTCCTC
	TCTTCTCTCTCCCTCGGTTACCCCCGGTTCCCTATGAGGTCATGCCACTGT
	TACCACGTTCCAGGCCCAGAGACGGAGGCAGGTTGGTCAAAGCCAGTCA
	CTCTCTGAACCCAGAGGTTGAGGAAGAGTGCATGCTGCGTGGAACGCTG
	GTCTTCCCCCATGGATGGCATGCTAGTTTCTCCAGCAAGCTGAGTCTCAT
	GTCCCCAAAGACGGGGACTTCCTGAGAAGCCTGGAGAGACAAGGGCTCC
	GTGGATGTCACTCTTAGGGAGGGTGTCCTGCAGCCCTCATTGACCTCCAC
	GACTAGGCTATGGTCTCCAGCCCCTCACAGCTCGTGGATAATTTGTGTTT
	CTTCGCTTTTGTTTTTTGTCTTTTCAAAGTGACTTTTTCCCCACTGGATTTC
	TAAGTTTCTCTTTGAAAATCAGTTCACTGGCAAATGGGACCTGCATCCTG
	ACCTGGCTGCCTGCATCAGGAGCGACACCAAACAGAGTGCGTGGGGATC
	CCCAATTGGCCCAGTGTCCCCCGGCCCTTCCTTAAGTCACACAAGCTCCC
	GTGTGGCTTTCGTGAGCATGGAGAACCTGTCCCCTGGTCTTAGAGAAAG
	CCAGCCATTCTGCCACCCTCTGTTTGTCTGGCAGACAGATTACCACACCG
	TGGCTGTCTTTCTAGCCAAAGCTTCCTCTCTCAACACCCATGAACGTCCA
	TGCTTCCTGTCTGAGCACTGAGGAGAACCCCAGCGGAGCTCATTGTTCAG
	TGCTGGAATACCCATCCCCCCTCCCGTTGATTATTTAGGGAGTGTCTGAT
	AATGCCAGGGGATACTCTGGGTGCTAGGGCGCAGAAGTACTTAAGAGCA
	AGTCCCAGCCTCAGGGGACTTATATGCCGGCGAGGAGAAAGCCAACAAA
	CCAATAAACTATGCACTGGTT
Modified	ATGGACCCCGGCCAGCAGCCCCCCCCCCAGCCCGCCCCCCAGGGCCAGG	25
Human YAP	GCCAGCCCCCCTCCCAGCCCCCCCAGGGCCAGGGCCCCCCCTCCGGCCC
(activated and	CGGCCAGCCCGCCCCCGCCGCCACCCAGGCCGCCCCCCAGGCCCCCCCC
without tag)	GCCGGCCACCAGATCGTGCACGTGCGCGGCGACTCCGAGACCGACCTGG
	AGGCCCTGTTCAACGCCGTGATGAACCCCAAGACCGCCAACGTGCCCCA
	GACCGTGCCCATGCGCCTGCGCAAGCTGCCCGACTCCTTCTTCAAGCCCC
	CCGAGCCCAAGTCCCACTCCCGCCAGGCCTCCACCGACGCCGGCACCGC
	CGGCGCCCTGACCCCCCAGCACGTGCGCGCCCACGCCTCCCCCGCCTCCC
	TGCAGCTGGGCGCCGTGTCCCCCGGCACCCTGACCCCCACCGGCGTGGT
	GTCCGGCCCCGCCGCCACCCCCACCGCCCAGCACCTGCGCCAGTCCTCCT
	TCGAGATCCCCGACGACGTGCCCCTGCCCGCCGGCTGGGAGATGGCCAA
	GACCTCCTCCGGCCAGCGCTACTTCCTGAACCACATCGACCAGACCACC
	ACCTGGCAGGACCCCCGCAAGGCCATGCTGTCCCAGATGAACGTGACCG
	CCCCCACCTCCCCCCCCGTGCAGCAGAACATGATGAACTCCGCCTCCGGC
	CCCCTGCCCGACGGCTGGGAGCAGGCCATGACCCAGGACGGCGAGATCT
	ACTACATCAACCACAAGAACAAGACCACCTCCTGGCTGGACCCCCGCCT
	GGACCCCCGCTTCGCCATGAACCAGCGCATCTCCCAGTCCGCCCCCGTGA
	AGCAGCCCCCCCCCCTGGCCCCCCAGTCCCCCCAGGGCGGCGTGATGGG
	CGGCTCCAACTCCAACCAGCAGCAGCAGATGCGCCTGCAGCAGCTGCAG
	ATGGAGAAGGAGCGCCTGCGCCTGAAGCAGCAGGAGCTGCTGCGCCAG
	GAGCTGGCCCTGCGCTCCCAGCTGCCCACCCTGGAGCAGGACGGCGGCA
	CCCAGAACCCCGTGTCCTCCCCCGGCATGTCCCAGGAGCTGCGCACCAT
	GACCACCAACTCCTCCGACCCCTTCCTGAACTCCGGCACCTACCACTCCC
	GCGACGAGTCCACCGACTCCGGCCTGTCCATGTCCTCCTACTCCGTGCCC
	CGCACCCCCGACGACTTCCTGAACTCCGTGGACGAGATGGACACCGGCG
	ACACCATCAACCAGTCCACCCTGCCCTCCCAGCAGAACCGCTTCCCCGAC
	TACCTGGAGGCCATCCCCGGCACCAACGTGGACCTGGGCACCCTGGAGG
	GCGACGGCATGAACATCGAGGGCGAGGAGCTGATGCCCTCCCTGCAGGA
	GGCCCTGTCCTCCGACATCCTGAACGACATGGAGTCCGTGCTGGCCGCC
	ACCAAGCTGGACAAGGAGTCCTTCCTGACCTGGCTG
Modified	ATGAACGTGCCCCTGGGCGGCATCTGGCTGTGGCTGCCCCTGCTGCTGAC	26
Mouse WNT2	CTGGCTGACCCCCGAGGTGTCCTCCTCCTGGTGGTACATGCGCGCCACCG
	GCGGCTCCTCCCGCGTGATGTGCGACAACGTGCCCGGCCTGGTGTCCCGC
	CAGCGCCAGCTGTGCCACCGCCACCCCGACGTGATGCGCGCCATCGGCC
	TGGGCGTGGCCGAGTGGACCGCCGAGTGCCAGCACCAGTTCCGCCAGCA
	CCGCTGGAACTGCAACACCCTGGACCGCGACCACTCCCTGTTCGGCCGC
	GTGCTGCTGCGCTCCTCCCGCGAGTCCGCCTTCGTGTACGCCATCTCCTC
	CGCCGGCGTGGTGTTCGCCATCACCCGCGCCTGCTCCCAGGGCGAGCTG
	AAGTCCTGCTCCTGCGACCCCAAGAAGAAGGGCTCCGCCAAGGACTCCA
	AGGGCACCTTCGACTGGGGCGGCTGCTCCGACAACATCGACTACGGCAT
	CAAGTTCGCCCGCGCCTTCGTGGACGCCAAGGAGCGCAAGGGCAAGGAC
	GCCCGCGCCCTGATGAACCTGCACAACAACCGCGCCGGCCGCAAGGCCG
	TGAAGCGCTTCCTGAAGCAGGAGTGCAAGTGCCACGGCGTGTCCGGCTC
	CTGCACCCTGCGCACCTGCTGGCTGGCCATGGCCGACTTCCGCAAGACC
	GGCGACTACCTGTGGCGCAAGTACAACGGCGCCATCCAGGTGGTGATGA
	ACCAGGACGGCACCGGCTTCACCGTGGCCAACAAGCGCTTCAAGAAGCC
	CACCAAGAACGACCTGGTGTACTTCGAGAACTCCCCCGACTACTGCATC
	CGCGACCGCGAGGCCGGCTCCCTGGGCACCGCCGGCCGCGTGTGCAACC
	TGACCTCCCGCGGCATGGACTCCTGCGAGGTGATGTGCTGCGGCCGCGG
	CTACGACACCTCCCACGTGACCCGCATGACCAAGTGCGAGTGCAAGTTC
	CACTGGTGCTGCGCCGTGCGCTGCCAGGACTGCCTGGAGGCCCTGGACG
	TGCACACCTGCAAGGCCCCCAAGTCCGCCGACTGGGCCACCCCCACC
Modified	ATGCGCCCCGCCCCCGCCCTGGCCCTGGCCGCCCTGTGCCTGCTGGTGCT	27
Human	GCCCGCCGCCGCCGCCGCCGCCGCCTACTTCGGCCTGACCGGCCGCGAG
WNT9B	GTGCTGACCCCCTTCCCCGGCCTGGGCACCGCCGCCGCCCCCGCCCAGGC
	CGGCGCCCACCTGAAGCAGTGCGACCTGCTGAAGCTGTCCCGCCGCCAG
	AAGCAGCTGTGCCGCCGCGAGCCCGGCCTGGCCGAGACCCTGCGCGACG
	CCGCCCACCTGGGCCTGCTGGAGTGCCAGTTCCAGTTCCGCCAGGAGCG
	CTGGAACTGCTCCCTGGAGGGCCGCACCGGCCTGCTGCAGCGCGGCTTC
	AAGGAGACCGCCTTCCTGTACGCCGTGTCCGCCGCCGCCCTGACCCACG
	CCCTGGCCCGCGCCTGCTCCGCCGGCCGCATGGAGCGCTGCACCTGCGA
	CGACTCCCCCGGCCTGGAGTCCCGCCAGGCCTGGCAGTGGGGCGTGTGC
	GGCGACAACCTGAAGTACTCCACCAAGTTCCTGTCCAACTTCCTGGGCCC
	CAAGCGCGGCTCCAAGGACCTGCGCGCCCGCGCCGACGCCCACAACACC
	CACGTGGGCATCAAGGCCGTGAAGTCCGGCCTGCGCACCACCTGCAAGT
	GCCACGGCGTGTCCGGCTCCTGCGCCGTGCGCACCTGCTGGAAGCAGCT
	GTCCCCCTTCCGCGAGACCGGCCAGGTGCTGAAGCTGCGCTACGACACC
	GCCGTGAAGGTGTCCTCCGCCACCAACGAGGCCCTGGGCCGCCTGGAGC
	TGTGGGCCCCCGCCAAGCCCGGCGGCCCCGCCAAGGGCCTGGCCCCCCG
	CCCCGGCGACCTGGTGTACATGGAGGACTCCCCCTCCTTCTGCCGCCCCT
	CCAAGTACTCCCCCGGCACCGCCGGCCGCGTGTGCTCCCGCGACTCCTCC
	TGCTCCTCCCTGTGCTGCGGCCGCGGCTACGACACCCAGTCCCGCATGGT
	GGTGTTCTCCTGCCACTGCCAGGTGCAGTGGTGCTGCTACGTGGAGTGCC
	AGCAGTGCGCCCAGCAGGAGCTGGTGTACACCTGCAAGCGC
WT YAP	GACTACAAAGACGATGACGATAAAGCAAGGCTCGAATCGGTACCTATG	28
FLAG Tag
Modified YAP	GACTACAAGGACGACGACGACAAGGCCCGCCTGGAGTCCGTGCCCATG	29
FLAG Tag
WT beta	GAGCAAAAGCTCATTTCTGAAGAGGACTTG	30
catenin myc
Tag
Modified beta	GAGCAGAAGCTGATCTCCGAGGAGGACCTG	31
myc FLAG
Tag

TABLE 2 — Combinations of liver regenerative factors VEGF

(e.g.

GF-1

Secreted

Beta-

EGF

HGF

P21

VEGF165)

IGF-1

IL-2 SP

EGF

STAT5B

catenin

YAP

Wnt9b

Wnt2

EGF

HGF

P21

VEGF

165

IGF-1

IL2 SP

Secreted

EGF

STAT

Beta-

catenin

YAP

Wnt9b

Wnt2

TABLE 3 — Combinations of sequences of liver regenerative factor mRNAs SEQ ID NO:

SEQ

NO:

TABLE 4 — Details of human tissue samples for histology evaluation.

Code	Age	Sex	Diagnosis	Child-Pugh Score
HH98	15 years	male	Donor liver	NA
HH104	11 years	male	Donor liver	NA
HH09	37 years	female	FNH/No chemo	NA
HH119	59 years	male	NASH cirrhosis	B
HH123	58 years	male	NASH cirrhosis	C
HH124	63 years	male	NASH cirrhosis	B
HH125	66 years	male	NASH cirrhosis	B
HH121	71 years	male	Alcoholic cirrhosis	C

TABLE 5 — Details of human tissue samples from cirrhotic and normal liver from which hepatocytes were recovered for gene and protein expression analysis.

Code	Age	Sex	Diagnosis	Child-Pugh Score
HH48	33 years	female	HCA “normal”	NA
HH55	61 years	male	ICCA “normal”	NA
HH16	44 years	male	Colon Ca “normal”	NA
HH36	47 years	female	EHE “normal”	NA
HH98	15 years	male	Donor liver “normal”	NA
HH32	71 years	female	NASH cirrhosis	B
HH72	48 years	male	Alcoholic cirrhosis	B
HH49	50 years	female	NASH cirrhosis	B
HH62	67 years	male	NASH cirrhosis	B
HH22	53 years	male	NASH cirrhosis	B
HH26	52 years	male	Alcoholic cirrhosis	C
HH29	68 yo	female	NASH cirrhosis	C
HH97	62 yo	female	NASH cirrhosis	C
HH123	58 yo	male	NASH cirrhosis	C

TABLE 7 — List of primary and secondary antibodies used in tissue immunostaining. Antigen

Antibody	Dilution	Cat. No.	Company	Retrieval
RFP	1:300	600-401-379S	Rockland	—
EpCAM	1:100	552370	BD Pharmingen	—
KRT7	1:1000	ab181598	Abcam	pH = 9 for 20
				min at 95° C.
				for human
				tissue
KDR	1:100	2479s	Cell signaling	pH = 9 for 20
				min at 95° C.
				for human
				tissue
HNF4a	1:200	ab201460	Abcam	—
CD26	1:100	AF954-SP	R&D	—
KRT19	1:100	602-670	Abbomax	—
YFP/GFP	1:300	A10262	Life	—
			technologies
GS	1:500	MA5-27749	Invitrogen	pH = 9 for 20
				min at 95° C.
				for human
				tissue
p21	1:500	ab188224	Abcam	—
CD31	1:100	557355	BD Pharmingen	—

TABLE 8 — List of conjugated antibodies used in flow cytometry.

Antibody	Dilution	Cat. No.	Company
APC anti-mouse CD45	1:100	103112	Biolegend
APC anti-mouse TER-119	1:100	116211	Biolegend
APC anti-mouse CD31	1:100	102409	Biolegend
APC anti-mouse/human	1:100	101211	Biolegend
CD11b
BV605 Rat Anti-Mouse CD26	1:100	BDB745125	BD OptiBuild
APC/Cyanine7 anti-mouse	1:100	118217	Biolegend
CD326 (EpCAM)
Rat anti-mouse CD16/32	1:50	553141	BD Pharmingen
(Fc Block)

TABLE 9 — Transgenic zebrafish lines used in this study.

Names used in this study	Official names (ZFIN database)	Allele #	Reference
Tg(Tp1:H2B-mCherry)	Tg(EPV.Tp1-Mmu.Hbb:hist2h2l-mCherry)	s939	[1]
Tg(fabp10a:CFP-NTR)	Tg(fabp10a:CFP-NTR)	s931	[1]
Tg(hs:sflt1)	Tg(hsp70l:flt1, cryaa:Cerulean)	bns80	[2]
Tg(Tp1:CreERT2)	Tg(EPV.Tp1-Mmu.Hbb:Cre-	s959	[1]
	ERT2, cryaa:mCherry)
Tg(hs:loxP-mCherry-	Tg(hsp701:LOXP-mCherry-LOXP-	mn32	[3]
loxP-hVEGFA)	Hsa.VEGFA, XIa.Cryg:EGFP)

TABLE 10 — List of conjugated antibodies used in western blot.

Antibody	Host	Company	Cat. No.	Dilution
HNF4A	Mouse	Abcam	Ab41898	1:1000
KDR	Rabbit	Cell	2479s	1:1000
		Signaling
CD31	Rabbit	Abcam	Ab32457	1:2500
KRT19	Mouse	Progen	61029	1:500
KRT7	Mouse	Novus	NBP2-44814	1:2000
		Biologicals
GAPDH	Mouse	Proteintech	60004-l	1:10000

TABLE 11 — List of human primers used for gene expression analysis in isolated human hepatocytes.

Gene	Primer Sequence	Amplicon length
hKRT7	F:	GGACATCGAGATCGCCACCT (SEQ ID NO: 32)	124
	R:	ACCGCCACTGCTACTGCCA (SEQ ID NO: 33)
h18s rRNA	F:	CTCAACACGGGAAACCTCAC (SEQ ID NO: 34)	110
	R:	CGCTCCACCAACTAAGAACG (SEQ ID NO: 35)
hKDR	F:	ACTTTGGAAGACAGAACCAAATTATCTC (SEQ ID NO: 36)	52
	R:	TGGGCACCATTCCACCA (SEQ ID NO: 37)
hAlbumin	F:	GTGAAACACAAGCCCAAGGCAACA (SEQ ID NO: 38)	116
	R:	TCCTCGGCAAAGCAGGTCTC (SEQ ID NO: 39)
hHNF4a	F:	GGTGTCCATACGCATCCTTGAC (SEQ ID NO: 40)	144
	R:	AGCCGCTTGATCTTCCCTGGAT (SEQ ID NO: 41)

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IPC · International Patent Classification

Section A — Human necessities

A61K31/198

Section C — Chemistry; metallurgy

C12N5/071
C12N15/88

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