GPO10, a collagen analog, effectively promotes activation of collagen‐bound pro‐Matrix‐Metalloproteinase‐2 in fibrotic liver tissue stimulating cell proliferation and migration

Abstract

Introduction: Matrix-metalloproteinase 2 (MMP-2) degrades denatured collagens and plays a key role in liver fibrogenesis/lysis. We previously showed that proMMP-2 binds to non-substrate ECM-collagens and is released and activated by the collagen-analog GPO10. The aim of this study was to elucidate cell-specific events initiated by GPO10-induced proMMP-2 activation. Methods: GPO10-induced proMMP-2 activation was shown by gel-zymography and substrate turnover. The hepatic stellate cell line CFSC was treated with nanomolar concentrations of pro- and activated MMP-2 with and without specific MMP-inhibition (ilomastat) and GPO10. DNA-synthesis was measured by [³H]-thymidine-incorporation. Additionally, a MMP-2-dependent migration assay was performed. Results: A 10-fold molar excess of GPO10 led to complete activation of proMMP-2 whereas GAP10 (control-peptide) had no effect. Active MMP-2 and GPO10 stimulated CFSC-proliferation up to 35% and 20% compared to max. stimulated cells, resp. GPO10–induced proliferation was completely blocked by ilomastat. MMP-2 and GPO10-activated proMMP-2 specifically induced strong cell migration. Conclusion: GPO10 activates collagen-bound proMMP-2 thus enhancing proliferation and migration of mesenchymal and hepatic stellate cells Local modulation of MMP-activity by GPO10 may open a new therapeutic approach in liver fibrosis.