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Marco Thilo Birke, Erion Lipo, Mehreen Adhi, Kerstin Birke, Rajendra Kumar-Singh; AAV Mediated Expression of Human PRELP inhibits Complement Activation, Choroidal Neovascularization and Deposition of Membrane Attack Complex in Mice. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1188.
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© ARVO (1962-2015); The Authors (2016-present)
Activation of complement has been previously associated with age related macular degeneration (AMD). We wished to investigate the potential use of human proline/arginine-rich end leucine-rich repeat protein (PRELP) as an inhibitor of complement deposition and laser induced choroidal neovascularization (CNV) in a murine model of AMD.
We constructed an AAV2/8 vector expressing human PRELP from a chicken beta actin promoter. PRELP-mediated inhibition of complement was examined using FACS lysis assays. PRELP-mediated inhibition of in vitro angiogenesis was measured using tube formation assays with HUVEC cells. AAV2/8 coding for PRELP was injected into the subretinal space of normal adult mice and the human PRELP protein was localized by immunocytochemistry. A separate group of injected mice were utilized for laser induced CNV. At 7 days post laser, eyes were harvested and stained for lectin or formation of the membrane attack complex (MAC).
In FACS lysis assays, PRELP reduced normal human serum (NHS) mediated lysis of Hepa-1c1c7 cells by 18.7+/-3.95%. Unexpectedly, in HUVEC tube formation assays, PRELP enhanced the formation of master junctions by 210% and enhanced formation of master segments and meshes by 230% and 290% respectively. Immunofluourescence analyses of PRELP expression in retinal sections identified the RPE and the photoreceptor outer segments as initial sites of AAV2/8 infection and expression, but indicated secretion of PRELP and it's subsequent migration to the inner retinal layers. PRELP reduced laser induced CNV by 60+/-13.1% and deposition of MAC at the site of CNV lesion by 25.5+/-12.3%.
PRELP is a potent inhibitor of complement and laser induced CNV in a model of AMD. Our results have implications for the development of complement inhibitors as a therapy for AMD.
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