title: Tissue inhibitor of metalloproteinases-3 peptides inhibit angiogenesis and choroidal neovascularization in mice.
abstract: Tissue inhibitors of metalloproteinases (TIMPs) while originally characterized as inhibitors of matrix metalloproteinases (MMPs) have recently been shown to have a wide range of functions that are independent of their MMP inhibitory properties. Tissue inhibitor of metalloproteinases-3 (TIMP-3) is a potent inhibitor of VEGF-mediated angiogenesis and neovascularization through its ability to block the binding of VEGF to its receptor VEGFR-2. To identify and characterize the anti-angiogenic domain of TIMP-3, structure function analyses and synthetic peptide studies were performed using VEGF-mediated receptor binding, signaling, migration and proliferation. In addition, the ability of TIMP-3 peptides to inhibit CNV in a mouse model was evaluated. We demonstrate that the anti-angiogenic property resides in the COOH-terminal domain of TIMP-3 protein which can block the binding of VEGF specifically to its receptor VEGFR-2, but not to VEGFR-1 similar to the full-length wild-type protein. Synthetic peptides corresponding to putative loop 6 and tail region of TIMP-3 have anti-angiogenic properties as determined by inhibition of VEGF binding to VEGFR-2, VEGF-induced phosphorylation of VEGFR-2 and downstream signaling pathways as well as endothelial cell proliferation and migration in response to VEGF. In addition, we show that intravitreal administration of TIMP-3 peptide could inhibit the size of laser-induced choroidal neovascularization lesions in mice. Thus, we have identified TIMP-3 peptides to be efficient inhibitors of angiogenesis and have a potential to be used therapeutically in diseases with increased neovascularization.
authors.affiliation: Department of Ophthalmology, Cole Eye Institute, Cleveland Clinic Lerner College of Medicine at Case Western Reserve University, Cleveland, Ohio, United States of America.
authors.forename: Jian Hua
authors.initials: JH
authors.lastname: Qi
authors.forename: Quteba
authors.initials: Q
authors.lastname: Ebrahem
authors.forename: Mariya
authors.initials: M
authors.lastname: Ali
authors.forename: Alecia
authors.initials: A
authors.lastname: Cutler
authors.forename: Brent
authors.initials: B
authors.lastname: Bell
authors.forename: Nicholas
authors.initials: N
authors.lastname: Prayson
authors.forename: Jonathan
authors.initials: J
authors.lastname: Sears
authors.forename: Vera
authors.initials: V
authors.lastname: Knauper
authors.forename: Gillian
authors.initials: G
authors.lastname: Murphy
authors.forename: Bela
authors.initials: B
authors.lastname: Anand-Apte
mesh_terms: D020533:Angiogenesis Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology*; D000818:Animals; D016570:Bruch Membrane / Q000098:blood supply* / Q000187:drug effects* / Q000293:injuries; D002478:Cells, Cultured; D020256:Choroidal Neovascularization / Q000209:etiology / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control*; D004730:Endothelium, Vascular; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D017075:Laser Coagulation / Q000009:adverse effects; D051379:Mice; D008810:Mice, Inbred C57BL; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D010766:Phosphorylation; D015398:Signal Transduction / Q000187:drug effects; D060327:Solid-Phase Synthesis Techniques; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D010455:Peptides; D019717:Tissue Inhibitor of Metalloproteinase-3; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2