1 September 2012
(GPO)10 , extracellular matrix, in vitro , matrixmetalloproteinase, proMMP-2, gelatinases, enzymatic activation, HT1080, hepatic stellate cellss, DMEM, Dulbecco's Modified Eagle's Medium, DQ-gelatin, dye-quenched gelatin, DSS, dextran sulfate sodium, ECM, extracellular matrix, FCS, fetal calf serum, GAP, H-Gly-Ala-Cys-(Gly-Ala-Pro)5-Gly-Phe-Hyp-Gly-Glu-Arg-(Gly-Ala-Pro)5-NH2 , (GPO)10, H-Gly-Cys-Hyp-(Gly-Pro-Hyp)10-Gly-Cys-Hyp-Gly-NH2 , HSC, hepatic stellate cell, HT1080, human fibrosarcoma cell line, MMP, matrixmetalloproteinase, PBS, phosphate buffered saline, PDGF-BB, platelet-derived growth factor subunit B, PI3-K, phosphoinositide 3-kinase
Diseases such as liver fibrosis and intestinal inflammation are characterized by accumulated components of the extracellular matrix (ECM). Given that fibrillar collagen structures were shown to serve as storage site for inactive proforms of matrixmetalloproteinases (MMPs), modulating this MMP-collagen interaction might offer a rational interventional (therapeutic) approach to enhance degradation of accumulated ECM. The synthetic triple helical collagen analogue (Gly-Pro-Hyp) 10 — (GPO) 10 — was shown to trigger release and enzymatic activation of collagen sequestered proMMP-2. In the presented study, we, for the first time, investigated how MMP-(GPO) 10 interaction impacts cellular responses in vitro. We found that recombinant proMMP-2 induced proliferation of hepatic stellate cells (HSC), which was enhanced after addition of (GPO) 10 reaching comparable levels following incubation with fully activated MMP-2. In addition, (GPO) 10 induced HSC migration similar to the platelet-derived growth factor subunit-B. Further, the MMP-2-dependent invasion of HT1080 fibrosarcoma cells through an ECM membrane was enhanced after addition of (GPO) 10. Since cellular proliferation and migration concomitant with matrix degradation is stimulated, we conclude that the MMP-(GPO) 10 interaction also functions in a physiological environment. Thus, a potential therapeutic effect of (GPO) 10 should be further tested in animal models for MMP-associated diseases such as colitis or fibrosis.