ABSTRACT
PURPOSE: To explore the in vivo anti-angiogenesis effects resulting from lentivirus-mediated RNAi of vascular endothelial growth factor (VEGF) in monkeys with iris neovascularization (INV). METHODS: Five specific recombinant lentiviral vectors for RNA interference, targeting Macaca mulatta VEGFA, were designed and the one with best knock down efficacy (LV-GFP-VEGFi1) in H1299 cells and RF/6A cells was selected by real-time PCR for in vivo use. A laser-induced retinal vein occlusion model was established in one eye of seven cynomolgus monkeys. In monkeys number 1, 3, and 5 (Group 1), the virus (1x10(8) particles) was intravitreally injected into the preretinal space of the animal's eye immediately after laser coagulation; and in monkeys number 2, 4, and 6 (Group 2), the virus (1x10(8) particles) was injected at 10 days after laser coagulation. In monkey number 7, a blank control injection was performed. In monkeys number 1 and 2, virus without RNAi sequence was used; in monkeys number 3 and 4, virus with nonspecific RNAi sequence was used; and in monkeys 5 and 6, LV-GFP-VEGFi1 was used. RESULTS: In monkey number 5, at 23 days after laser treatment, no obvious INV was observed, while fluorescein angiography of the iris revealed high fluorescence at the margin of pupil 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 iridic vessels all significantly varied: between monkey number 5 and number 3 (36.01+/-4.49/mm(2) versus 48.68+/-9.30/mm(2), p=0.025), between monkey number 3 and monkey number 7 (48.68+/-9.30/mm(2) versus 74.38+/-9.23/mm(2), p=0.002), and between monkey number 5 and number 7 (36.01+/-4.49/mm(2) versus 74.38+/-9.23/mm(2), p<0.001). CONCLUSIONS: Lentivirus-mediated RNAi of VEGF may be a new strategy to treat iris neovascularization, while further studies are needed to investigate the long-term effect.
Subject(s)
Iris/blood supply , Iris/pathology , Lentivirus/metabolism , Neovascularization, Pathologic/therapy , RNA Interference , Vascular Endothelial Growth Factor A/metabolism , Animals , Disease Models, Animal , Fluorescein Angiography , Green Fluorescent Proteins/metabolism , Haplorhini , Immunohistochemistry , Neovascularization, Pathologic/diagnosis , RNA, Small Interfering/metabolismABSTRACT
A total of 67 eyes of 67 patients that have undergone vitrectomy and silicone oil tamponade for rhegmatogenous retinal detachment were included in this prospective clinical study. We measured the axial length (AL) and the anterior chamber depth (ACD) with A-mode ultrasonography and IOL Master before and after removal of silicone oil. Phase refractive indices of silicone oil at different wavelengths from 400 nm to 1000 nm were measured with an Abbe refractometer and a series of equations were derived to adjust the AL values obtained with the IOL Master in silicone-filled eyes. The instruments showed good agreement between the AL values measured before and after removal of silicone oil. The ACD values obtained with A-mode ultrasonography were lower than the values obtained with the IOL Master (p < 0.01). The AL values obtained with the IOL Master after removal of silicone oil were lower than the values obtained preoperatively (p < 0.01). The AL values obtained with the IOL Master after adjustment are more accurate than A-mode ultrasonography in silicone-filled eyes.
