MT1-MMP-mediated cleavage of decorin in corneal angiogenesis.
OBJECTIVE
Decorin has been shown to have antiangiogenic properties. In this study, we evaluate the involvement of membrane type 1-matrix metalloproteinase (MT1-MMP), a proangiogenic enzyme, in decorin cleavage in the cornea.


METHODS
MT1-MMP expression was confirmed immunohistochemically in keratocytes and immortalized corneal fibroblast cell lines. Corneal micropockets of bFGF were used to assess the expression of decorin and MT1-MMP. Western blotting was used to evaluate decorin degradation by MT1-MMP. Aortic ring tube formation assays were used to assay the inhibitory effect of decorin and stimulatory effect of MT1-MMP on vascular endothelial cells in vitro.


RESULTS
We show that MT1-MMP expression is upregulated following bFGF pellet implantation in the cornea in vivo, and that MT1-MMP cleaves decorin in a time- and concentration-dependent manner in vitro. Furthermore, the addition of MT1-MMP reduces the inhibitory effects of decorin on aortic ring tube formation in vitro. Cleavage of decorin by MT1-MMP-deficient corneal cell lysates is diminished relative to that by wild-type corneal cell lysates, and an MT1-MMP knockin restores decorin processing in vitro.


CONCLUSIONS
The proangiogenic role of MT1-MMP in the cornea may be mediated, in part, by facilitated cleavage of corneal decorin.
Mimura
Tatsuya
T
Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, IL 60612, USA.
Han
Kyu Yeon
KY
Onguchi
Tatsuya
T
Chang
Jin-Hong
JH
Kim
Tae-im
TI
Kojima
Takashi
T
Zhou
Zhongjun
Z
Azar
Dimitri T
DT
D000818:Animals; D001011:Aorta / Q000201:enzymology; D002460:Cell Line; D003315:Cornea / Q000201:enzymology*; D016510:Corneal Neovascularization / Q000139:chemically induced / Q000201:enzymology*; D017077:Culture Media, Conditioned / Q000378:metabolism; D058575:Decorin; D004151:Dipeptides / Q000494:pharmacology; D004195:Disease Models, Animal; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2; D007700:Kinetics; D053511:Matrix Metalloproteinase 14 / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011480:Protease Inhibitors / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D046509:Tissue Culture Techniques; D014162:Transfection
D017077:Culture Media, Conditioned; C546878:DCN protein, human; C546879:Dcn protein, mouse; D058575:Decorin; D004151:Dipeptides; D016326:Extracellular Matrix Proteins; D061965:Matrix Metalloproteinase Inhibitors; C507096:Mmp14 protein, mouse; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D011480:Protease Inhibitors; D011509:Proteoglycans; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14