A novel peptide binding to the C-terminal domain of connective tissue growth factor for the treatment of bleomycin-induced pulmonary fibrosis

Abstract

Using phage display technology, 810A was developed as a novel peptide, which binds to the C-terminal domain of connective tissue growth factor (CTGF). The present study investigated whether this peptide could neutralize CTGF and block its biological activity as a specific strategy to treat pulmonary fibrosis in vitro and in vivo. Human bronchial epithelial (16HBE) cells were treated with CTGF to evaluate the anti-CTGF capability of 810A using MTT testing, cell migration assays, and Western blotting. The action of 810A in mouse models of bleomycin (BLM)-induced pulmonary fibrosis was evaluated according to leukocyte counts in bronchoalveolar lavage fluid (BALF), hydroxyproline content, and pathological analysis. Treatment with 810A resulted in inhibition of cell proliferation, migration, and increases in transforming growth factor-beta (TGF-β) and α-Smooth muscle actin (α-SMA) caused by CTGF treatment in vitro. In vivo, 810A significantly alleviated fibrosis in the pulmonary index. Inflammation was inhibited, as inferred by the reduced number of leukocytes in BALF. Pathophysiological analysis indicated that hydroxyproline content, alveolar enlargement, interstitial inflammatory infiltration, and collagen deposition were reduced. Collectively, these data suggest that 810A may potentially be a CTGF-specific, small-molecule antagonist, providing a new method for prevention and treatment of pulmonary fibrosis.