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L. Luo, N. Singh, H. Uehara, J. Simonis, Y. Qazi, E. Pearson, M. Dixit, S. A. Molokhia, B. K. Ambati; Subretinal Injection of Adeno-associated Virus Expressing shRNA Against Soluble VEGF Receptor 1 (aav.shRNA.sflt-1) Induces Choroidal Neovascularization in Mice. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2054.
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Vascular endothelial growth factor (VEGF) is known to play a central role during choroidal neovascularization. VEGF binding to an appropriate receptor like Flt is the main mitogen for endothelial cells and a strong inducer of angiogenesis. A soluble form of VEGF receptor, sFlt-1, specifically binds VEGF and inhibits its activity. It is probable that suppression of sFlt-1 could unmask activity of VEGF as a stimulator of inflammation and angiogenesis. In this study, we used small interfering RNA directed against sFlt-1 to investigate the role of sFlt-1 in choroid neovascularization.
An adeno-associated viral vector encoding small RNA interference-mediated soluble human VEGF receptor-1 (aav.siRNA.sFlt-1) was injected into the subretinal space of mice. PBS and recombinant viral vectors expressing the Green fluorescent protein (GFP) without siRNA.sFlt-1 were used as controls. CNV was observed by the combination of fluorescein angiography (FA) and indocyanine green (ICG) angiography using the Heidelberg Retina Angiograph. Four and seven weeks later, CNV was evaluated by fluorescein angiographic choroidal flat-mount image analysis. The expression of GFP was analyzed in retinal sections by direct fluorescence imaging.
Injection of vector expressing sFlt-1 resulted in an increase in the area of CNV by 3.7 times relative to control (P < 0.05). In addition to large CNV, occult CNV could be seen with ICG but not with FA when FA and ICG were performed concurrently.
AAV.siRNA.sFlt-1 delivery into the subretinal space induces CNV in mice. These data suggest that sFlt-1 plays an important role in preventing CNV in mouse.
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