Inhibition of neointimal hyperplasia by blocking alpha V beta 3 integrin with a small peptide antagonist GpenGRGDSPCA.

Abstract

Neointimal hyperplasia is a leading cause of restenosis after vascular procedures. Recent findings showed that smooth muscle cell (SMC) migration from the media into the neointima is a critical step in the development of the hemodynamically compromising neointimal lesion. Moreover, integrins are believed to play a role in SMC motility. Therefore we studied the role of one ubiquitous integrin, alpha V beta 3, in SMC migration. Transwell assay was used to study in vitro migration of human and rabbit SMCs after stimulation with platelet-derived growth factor (PDGF). A neutralizing monoclonal antibody to alpha V beta 3, LM609, and a specific arginine-glycine-aspartic acid (RGD) antagonist, GpenGRGDSPCA, were used in the migration assay to inhibit alpha V beta 3-mediated SMC migration. In addition, GpenGRGDSPCA was administered locally to rabbit carotid artery after balloon angioplasty to determine the effect of blocking alpha V beta 3 on neointimal hyperplasia. We showed that PDGF-induced human SMC migration is mediated by the alpha V beta 3 integrin by use of LM609 to inhibit migration and that SMC migration is RGD dependent by use of GpenGRGDSPCA to inhibit migration. We have also inhibited rabbit SMC migration with GpenGRGDSPCA to demonstrate the cross-species preservation of the RGD peptide sequence in SMC mortality. Finally, when we administered GpenGRGDSPCA locally to rabbit carotid artery after balloon angioplasty, there was a statistically significant reduction in neointimal lesion formation compared with arteries administered an inactive peptide or saline solution. We have demonstrated the important role of the alpha V beta 3 integrin in SMC migration in vitro and in neointimal hyperplasia in vivo.