Methods for identifying agents for preventing or treating proliferative diseases, and for inhibiting extracellular matrix or α1 type IV collagen

A method of identifying an agent effective in preventing and/or treating a proliferative disease causing sclerosis, comprisingncontacting a test agent with a biological sample;determining the level of expression of at least one substance selected from the group consisting of Smad1 and phosphorylated Smad1 in the biological sample in comparison to the level of expression of the substance in a control sample;wherein a decrease in expression of Smad1 or phosphorylated Smad1 in comparison to the expression level of the substance in the control sample indicates the agent is effective in the prevention and/or treatment of proliferative diseases causing sclerosis.

A method of identifying an agent effective in inhibiting the increase of extracellular matrix, comprisingncontacting a test agent with a biological sample;determining the level of expression of at least one substance selected from the group consisting of Smad1 and phosphorylated Smad1 in the biological sample in comparison to the level of expression of the substance in a control sample;wherein a decrease in expression of Smad1 or phosphorylated Smad1 in comparison to the expression level of the substance in the control sample indicates the agent is effective in inhibiting the increase of extracellular matrix.

A method of identifying substances effective in inhibiting the expression of α1 type IV collagen, comprisingncontacting a test agent with a biological sample;determining the level of expression of at least one substance selected from the group consisting of Smad1 and phosphorylated Smad1 in the biological sample in comparison to the level of expression of the substance in a control sample,wherein a decrease in expression of Smad1 or phosphorylated Smad1 in comparison to the expression level of the substance in the control sample indicates the agent is effective in inhibiting the expression of α1 type IV collagen.

The method of any one of claims 1, 2, or 3, wherein the biological sample is selected from the group consisting of renal tissue sections, blood, sera and urine.

The method of any one of claims 1, 2, or3, wherein the biological sample is selected from mesangial cells.

The method of any one of claims 1, 2, or3, wherein the level of expression is measured at the nucleic acid level or the protein level.

A method of identifying an agent effective in preventing and/or treating a proliferative disease causing sclerosis, or effective in inhibiting the increase of extracellular matrix, or effective in inhibiting the expression of α1 type IV collagen comprisingncontacting a test agent with a biological sample;determining the level of expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1 in the biological sample in comparison to the level of expression of the substance in a control sample,wherein a decrease in expression of STAT3. phosphorylated STAT3, Smad1 or phosphorylated Smad1 in comparison to the expression level of the substance in the control sample indicates the agent is effective in the prevention and/or treatment of proliferative diseases causing sclerosis, or effective in inhibiting the increase of extracellular matrix or effective in inhibiting the expression of α1 type IV collagen,and wherein the level of expression of STAT3, phosphorylated STAT3, Smad1 or phosphorylated Smad1 at the nucleic acid level is measured using primer pairs selected from SEQ ID NOS: 21 and 22, or SEQ ID NOS: 5 and 6.

The method of any one of claims 1, 2, or 3, wherein the level of expression of, Smad1 or phosphorylated Smad1 at the protein level is measured by Western Blotting, ELISA or immunohistochemical analysis.