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S.J. Reich, A. Kuroki, J. Fosnot, X.Y. Yang, A.M. Maguire, J. Bennett, M.J. Tolentino; VEGF Directed siRNA Inhibits Experimental Choroidal Neovascularization . Invest. Ophthalmol. Vis. Sci. 2003;44(13):578.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Gene silencing of angiogenic factors involved in pathologic neovascularization may be an important therapeutic anti-angiogenic strategy. The purpose of this study is to determine if murine vascular endothelial growth factor (VEGF)-directed siRNA will inhibit experimental laser induced choroidal neovascularization (CNV) in mice. Methods:Adult female C57BL/6 mice were laser photocoagulated using an 810 nm diode laser (OcuLight Six; IRIS Medical, Mountain View, CA). Three laser spots were applied in both eyes. One day post-treatment, subretinal injections were performed unilaterally to deliver siRNA that targets mouse VEGF. The siRNA was conjugated with transit TKO transfection reagent (Mirus) and mixed with recombinant adenovirus (rAdenovirus). Contralateral eyes received subretinal injections of siRNA targeting GFP and served as controls. This siRNA was also conjugated with transit TKO reagent (Mirus) and mixed with rAdenovirus. Two weeks following laser treatment, animals were perfused with high molecular weight FITC-Dextran, and choroidal flat mounts were prepared. Choroidal flat mounts were photographed and analyzed microscopically. The area of CNV was measured with Openlab software (Improvision, Boston, MA). Results: Eyes treated with VEGF directed siRNA showed a statistically significant decrease in the size of the choroidal neovasculariation as compared with the control eye. Average CNV area in eyes treated with siRNA directed to GFP was 0.08 mm2. In the murine VEGF directed siRNA treated eyes average CNV area was 0.03. mm 2. The difference was statistically significant ( p< 0.00004). Conclusion: Gene silencing with siRNA directed towards VEGF can significantly inhibit neovascularization in the murine laser induced CNV model. This experiment helps validate the strategy of using siRNA directed towards VEGF as an anti-angiogenic treatment.
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