Lentivirus-mediated RNA interference of vascular endothelial growth factor in monkey eyes with iris neovascularization

[摘要] Purpose: To explore the in vivoanti-angiogenesis effects resulting from lentivirus-mediated RNAi ofvascular endothelial growth factor (VEGF) in monkeys with irisneovascularization (INV). Methods: Five specific recombinantlentiviral vectors for RNA interference, targeting Macaca mulattaVEGFA, were designed and the one with best knock down efficacy(LV-GFP-VEGFi1) in H1299 cells and RF/6A cells was selected byreal-time PCR for in vivo use. A laser-induced retinal vein occlusionmodel was established in one eye of seven cynomolgus monkeys. Inmonkeys number1, 3, and 5 (Group 1), the virus (1×108particles) was intravitreally injected into the preretinal space of theanimal's eye immediately after laser coagulation; and in monkeys number2, 4, and 6 (Group 2), the virus (1×108 particles) wasinjected at 10 days after laser coagulation. In monkey number 7, ablank control injection was performed. In monkeys number 1 and 2, viruswithout RNAi sequence was used; in monkeys number 3 and 4, virus withnonspecific RNAi sequence was used; and in monkeys 5 and 6,LV-GFP-VEGFi1 was used. Results: In monkey number 5, at 23 daysafter laser treatment, no obvious INV was observed, while fluoresceinangiography of the iris revealed high fluorescence at the margin ofpupil and point posterior synechiae. At 50 days after laser treatment,only a slight ectropion uvea was found. However, in the other eyes,obvious INV or hyphema was observed. The densities of new iridicvessels all significantly varied: between monkey number 5 and number 3(36.01±4.49/mm2 versus 48.68±9.30/mm2, p=0.025),between monkey number 3 and monkey number 7 (48.68±9.30/mm2versus 74.38±9.23/mm2, p=0.002), and between monkey number 5and number 7 (36.01±4.49/mm2 versus 74.38±9.23/mm2,p<0.001). Conclusions: Lentivirus-mediated RNAi ofVEGF may be a new strategy to treat iris neovascularization, whilefurther studies are needed to investigate the long-term effect.

[学科分类] 生物化学/生物物理