Functional characterization of neostatins, the MMP-derived, enzymatic cleavage products of type XVIII collagen.
Several anti-angiogenic factors are derived from proteolytic processing of large molecules including endostatin from type XVIII collagen and angiostatin from plasminogen. In previous studies we showed that neostatin-7, the C-terminal 28kDa endostatin-spanning proteolytic fragment, is generated from the proteolytic action of matrix metalloproteinase matrilysin (MMP)-7 on type XVIII collagen. Now, we report a second member of the neostatin family of proteins, neostatin-14. Given the small quantities of neostatin-7 and -14 generated by the breakdown of naturally occurring collagen XVIII (using MMP-7 and -14, respectively), we used two other approaches to characterize the anti-angiogenic properties of these molecules: murine recombinant neostatin in vitro, and gene therapy. We demonstrate that murine recombinant neostatin-7 inhibits calf pulmonary artery endothelial cell proliferation and that microinjection of neostatin-7 and neostatin-14 naked DNA into the corneal stroma of mice results in significant reduction of basic fibroblast growth factor-induced corneal neovascularization. These results provide supportive evidence of the possible anti-angiogenic effect of neostatins.
Corneal, External Disease and Refractive Surgery Service, Massachusetts Eye and Ear Infirmary and the Schepens Eye Research Institute, Harvard Medical School, 243 Charles Street, Boston, MA 02114, United States.
Jin-Hong
JH
Chang
Joel A D
JA
Javier
Gene-Yuan
GY
Chang
Hailton B
HB
Oliveira
Dimitri T
DT
Azar
D000595:Amino Acid Sequence; D020533:Angiogenesis Inhibitors / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects; D043170:Collagen Type XVIII / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D043169:Endostatins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects*; D016222:Fibroblast Growth Factor 2 / Q000037:antagonists & inhibitors; D015316:Genetic Therapy; D020783:Matrix Metalloproteinase 7 / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic / Q000235:genetics; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000166:cytology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
D020533:Angiogenesis Inhibitors; D043170:Collagen Type XVIII; D043169:Endostatins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C503330:neostatin-14 protein, mouse; C503329:neostatin-7 protein, mouse; D016222:Fibroblast Growth Factor 2; D020783:Matrix Metalloproteinase 7