CTGF and SMADs, maintenance of scleroderma phenotype is independent of SMAD signaling

In normal adult fibroblasts, transforming growth factor-beta (TGF beta) induces the expression of connective tissue growth:factor (CTGF). CTGF independently promotes fibroblast proliferation and matrix deposition, and in acute models of fibrosis promotes cell proliferation and collagen deposition acting synergistically with TGF-beta, In contrast to normal fibroblasts, fibroblasts cultured from fibrotic tissues express high basal levels of CTGF, even in the absence of added TGF beta, Induction of transcription by TGF beta requires the action of SMAD proteins. In this report we have investigated the role of SMADs in the TGF beta -induction of CTGF in normal fibroblasts and in the elevated levels of CTGF expression found in dermal fibroblasts cultured from lesional areas of patients with scleroderma, a progressive fibrotic disorder that can affect all organs of the body. We have identified a functional SMAD binding site in the CTGF promoter. TGF beta -induction of CTGF is dependent on SMAD3 and SMAD4:but not SMAD2 and is p300-independent, However, mutation of the SMAD binding site does not reduce the high level of CTGF promoter activity observed in dermal fibroblasts cultured from lesional areas of scleroderma patients, Conversely, the previously termed TGF beta RE in the CTGF promoter is required for basal CTGF promoter activity in normal fibroblasts and for the:elevated level of CTGF promoter activity in scleroderma fibroblasts, Thus, the maintenance of the fibrotic phenotype in scleroderma fibroblasts, as visualized by: excess CTGF expression, appears to be independent of SMAD-dependent TGF beta signaling. Furthermore, given CTGF's activities, the high level of CTGF expression observed in scleroderma lesions may contribute to the excessive scarring observed in this disorder.