Identification of multiple, differentially expressed messenger RNAs in dermal fibroblasts from patients with systemic sclerosis

Objective. To simultaneously identify several genes whose expression is alt ered in dermal fibroblasts from patients with systemic sclerosis (SSc), Methods. Total RNA was prepared from fibroblasts derived from clinically af fected and unaffected skin of patients with SSc. The RNA samples were analy zed using differential-display reverse transcription-polymerase chain react ion (DDRT-PCR), Complementary DNA (cDNA) fragments corresponding to differe ntially expressed messenger RNAs (mRNAs) were eluted, cloned, and sequenced , The differential expression of the corresponding mRNAs was confirmed by r ibonuclease protection assay. Results. We identified 21 differentially expressed mRNAs, Their correspondi ng cDNAs were sequenced and the sequences obtained were compared with those of known genes entered into the EMBL/GenBank database. Three of the sequen ces corresponded to transcripts of yet-unidentified genes. Some of the mRNA s shared partial homology with extracellular matrix components, cellular re ceptors, enzymes, and nuclear factors. Others corresponded to known mRNAs s uch as those of fibronectin, fibronectin receptor, laminin receptor homolog , beta-tubulin, insulin-like growth factor binding protein 5, KIAA0179 prot ein, and protease nexin 1, Conclusion. The application of DDRT-PCR to scleroderma research has identif ied many mRNAs whose altered expression in scleroderma has not yet been des cribed, thus providing ne cv information for further investigation and pote ntial targets for the development of novel therapies.