The role of integrin alpha5beta1 in the regulation of corneal neovascularization.
Integrins are transmembrane receptor proteins critical for growth and stabilization of vessels, but the mechanisms by which integrin activities are involved in neoangiogenesis of the eye remain unclear. Specific inhibitors to fibronectin receptor integrin alpha(5)beta(1) impeded pathological neovascularization in vivo. Our objective was to determine whether alpha(5)beta(1) plays a role in ocular angiogenesis, and whether a novel alpha(5)beta(1)-inhibiting small molecule is able to reduce angiogenesis in a model of inflammatory corneal neovascularization. Corneal neovascularization was induced in C57Bl/6 mice by NaOH-application and debridement of the limbal epithelium. Mice were randomized into six groups receiving either no treatment, or intraperitoneal osmotic pumps delivering three different doses of integrin antagonist or control substance on day 10 after scraping. In order to quantify the neovascular response, flatmounts were stained with FITC-CD31. Integrin alpha(5) expression was determined by immunohistochemistry and quantified by semiquantitative western blot analysis. Influence of integrin antagonist treatment on the mRNA expression of VEGF, bFGF and integrin alpha(5) was quantified by real-time RT-PCR. Vascularized corneas demonstrated a strong up-regulation of integrin alpha(5) within affected areas. Animals treated systemically with alpha(5)beta(1)-inhibiting small molecule showed a significant inhibition and regression of corneal neovascularization. PCR analysis evinced a significant up-regulation of VEGF and integrin alpha(5) mRNA levels in injured animals compared to controls, and a significant reduction of integrin alpha(5) mRNA in substance-treated animals compared to control substance, but no significant differences of bFGF levels in all groups. Western blot analysis of integrin alpha(5)beta(1) protein expression showed a trend towards up-regulation in injured animals, both control substance-treated and those treated with the alpha(5)beta(1)-inhibiting small molecule. Systemic delivery of an alpha(5)beta(1)-inhibiting small molecule inhibits and regresses corneal neovascularization induced by mechanical-alkali burn corneal injury. These results suggest an essential role for the integrin alpha(5)beta(1) in pathological neovascular processes of the cornea. Integrin alpha(5)beta(1) inhibitors could become a new approach for treatment of neovascularization in the eye.
Department of Vitreoretinal Surgery, Center of Ophthalmology, University of Cologne, Kerpener Strasse 62, 50924 Cologne, Germany.
Philipp S
PS
Muether
Susanne
S
Dell
Norbert
N
Kociok
Grit
G
Zahn
Roland
R
Stragies
Doerte
D
Vossmeyer
Antonia M
AM
Joussen
D020533:Angiogenesis Inhibitors / Q000097:blood / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D015153:Blotting, Western; D003315:Cornea / Q000378:metabolism; D016510:Corneal Neovascularization / Q000139:chemically induced / Q000188:drug therapy / Q000503:physiopathology* / Q000517:prevention & control; D004305:Dose-Response Relationship, Drug; D005260:Female; D039081:Integrin alpha5beta1 / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction / Q000379:methods; D011759:Pyrrolidines / Q000097:blood / Q000494:pharmacology / Q000627:therapeutic use; D012972:Sodium Hydroxide
D020533:Angiogenesis Inhibitors; D039081:Integrin alpha5beta1; C524156:JSM5562; D011759:Pyrrolidines; D012972:Sodium Hydroxide