April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
AAV Mediated Expression of Human PRELP inhibits Complement Activation, Choroidal Neovascularization and Deposition of Membrane Attack Complex in Mice
Author Affiliations & Notes
  • Marco Thilo Birke
    Ophthalmology, Tufts University, Boston, MA
  • Erion Lipo
    Ophthalmology, Tufts University, Boston, MA
  • Mehreen Adhi
    Ophthalmology, Tufts University, Boston, MA
  • Kerstin Birke
    Ophthalmology, Tufts University, Boston, MA
  • Rajendra Kumar-Singh
    Ophthalmology, Tufts University, Boston, MA
  • Footnotes
    Commercial Relationships Marco Birke, None; Erion Lipo, None; Mehreen Adhi, None; Kerstin Birke, None; Rajendra Kumar-Singh, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1188. doi:
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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)

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Abstract

Purpose: 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.

Methods: 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).

Results: 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%.

Conclusions: 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.

Keywords: 412 age-related macular degeneration  
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