Recognition Of Human Connective Tissue Growth Factor Gene And Its Promoter Activity And Gene Expression By Synthetic Pyrrole-imidazole Polyamide

Antisense technology is frequently being used for gene therapy. Antisense drugs is composed of antisense DNA, antisense RNA and ribozymes. However, these compounds are degraded easily by nucleases and require drug-delivery systems for sufficient distribution into organs. Therefore, suitable chemical modifications or drug delivery systems are necessary for their therapeutic application. Pyrrole-imidazole polyamides (PIP) are small synthetic molecules composed of the aromatic rings of the N-methylpyrrole and N-methylimidazole amino acid. Synthetic polyamides can bind to specific nucleotide sequences in the minor groove of double-helical DNA with high affinity and specificity and influence gene expression. PIP are chemical compounds that are completely resistant to nucleases and do not require particular delivery systems. PIP will be more feasible gene-silencing medicines. Connective tissue growth factor (CTGF) is recently thought to be the major pathogenic factor in the development of fibrotic disorders. As a downstream mediator of transforming growth factor beta (TGF-β),CTGF acts to promote fibroblast proliferation, DNA synthesis, ECM production, and its overproduction is proposed to play a major role in pathways that lead to fibrosis. CTGF mRNA has been shown to be overexpressed in the extracapillary and severe mesangial proliferative lesions of many progressive renal disease. Hence, CTGF has implicated in the pathogenesis of renal fibrosis. To develop a novel gene-silencing agent for the treatment of progressive renal disease, we designed and synthesized a PIP targeting human CTGF (hCTGF) promoter, and then examined its distribution and effects on hCTGF gene expression in human mesangial cells (HMCs). The present study includes three parts.PartⅠ: Specific recognition and cellular distribution of PIP...