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W. W. Hauswirth, R. Miller, J. Pang, V. Choido, A. Abraham, P. Pechan, H. Rubin, S. Wadsworth, Q. Li; Minimal Effective Dosing of a Novel Recombinant Form of VEGF Soluble Receptor 1 (sFlt-1) Delivered via AAV Vector in Inhibiting Choroidal Neovascularization in a Murine Model. Invest. Ophthalmol. Vis. Sci. 2008;49(13):1375.
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Choroidal neovascularization (CNV) occurs in variety of chorioretinal diseases, including age-related macular degeneration (AMD) and is the major cause of severe visual loss in patients with AMD. VEGF is known to be a critical stimulus for CNV. In a previous study, we showed that AAV-mediated delivery of a novel genetically engineered soluble form of VEGF receptor 1 (sFLT01) effectively inhibited CNV in an experimental animal model. In this study, we further investigated the minimal effective dose of the vector as a function of either intravitreal or subretinal routes of vector delivery in the mouse laser induced CNV model.
The cDNA for sFLT01 was packaged into an AAV serotype-2 vector and the vector serially diluted in saline ranging from full dose (~5X108 vg/eye) to 1000 fold dilution (~5X105 vg/eye). Each dilution was injected either intravitreously or subretinally into the right eyes of 10-15 adult C57BL/6 mice. 532nm laser photocoagulation was performed 4-6 weeks after injection to focally rupture Bruch’s membrane in both eyes (4-5 sites each eye). The development of CNV at each laser burn was evaluated 14 days later by fluorescein angiographic choroidal flat-mount image analysis, as well as by histology of transverse retinal sections. The expression of the sFlt01 in the retina was confirmed by immunocytochemistry.
sFLT01was primarily expressed in the retinal ganglion cells when the AAV2-CBA-sFLT01 vector was injected intravitreously, and in photoreceptors and RPE cells when injected subretinally. The minimum effective dose was approximately 4X107 vg/eye when the vector was injected intravitreously, and resulted in about 40% reduction of the CNV area per laser burn. In contrast, minimum effective dose for subretinal vector was more than ten fold lower (< 4X106 vg/eye) than for intravitreal vector.
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