title: Antiangiogenic and antivascular effects of a recombinant tumstatin-derived peptide in a corneal neovascularization model.
abstract: Tumstatin, a cleavage fragment of collagen IV, is a potent endogenous inhibitor of angiogenesis. Tumstatin-derived peptide T8 possesses all angiostatic properties of full-length tumstatin and indirectly suppresses tumor growth. The potential of T8 to block pathological angiogenesis in the eye has not been explored yet. Here we assess antiangiogenic effects of a recombinant T8 peptide in rabbit corneal neovascularization models. The fusion protein consisting of T8 and thioredoxin was synthesized in a highly efficient Escherichia coli expression system, isolated using ion-exchange chromatography and cleaved with TEV (tobacco etch virus) protease. The target peptide was purified on an anion-exchange resin and by reversed phase high-performance liquid chromatography. The recombinant peptide suppressed the proliferation of basic fibroblast growth factor-induced SVEC-4-10 endothelial cells (simian virus 40-immortalized murine endothelial cells) and inhibited tube formation in these cells in a dose-dependent manner. In rabbit corneal neovascularization models T8 demonstrated the ability to prevent pathological angiogenesis (when injected simultaneously with the induction of neovascularization) and, moreover, to promote the regression of newly-formed blood vessels (when injected on day 8 after angiogenesis stimulation). Our results suggest that T8 may have a therapeutic potential in the treatment of ocular neovascular diseases.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117997, Russia. esipov@ibch.ru
authors.forename: Roman
authors.initials: R
authors.lastname: Esipov
authors.forename: Ksenia
authors.initials: K
authors.lastname: Beyrakhova
authors.forename: Vera
authors.initials: V
authors.lastname: Likhvantseva
authors.forename: Evgenia
authors.initials: E
authors.lastname: Stepanova
authors.forename: Vasily
authors.initials: V
authors.lastname: Stepanenko
authors.forename: Maria
authors.initials: M
authors.lastname: Kostromina
authors.forename: Yulia
authors.initials: Y
authors.lastname: Abramchik
authors.forename: Anatoly
authors.initials: A
authors.lastname: Miroshnikov
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000737:chemistry / Q000627:therapeutic use*; D024141:Collagen Type IV / Q000737:chemistry / Q000627:therapeutic use*; D016510:Corneal Neovascularization / Q000188:drug therapy*; D042783:Endothelial Cells / Q000187:drug effects; D051379:Mice; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D010446:Peptide Fragments / Q000627:therapeutic use*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000627:therapeutic use*
chemical_list: D001324:Autoantigens; D024141:Collagen Type IV; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C057112:type IV collagen alpha3 chain