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

Yuan MK, Tao Y, Yu WZ, Kai W, Jiang YR - Mol. Vis. (2010)

Bottom Line: At 50 days after laser treatment, only a slight ectropion uvea was found.However, in the other eyes, obvious INV or hyphema was observed.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, People's Hospital, Peking University, Beijing, China.

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.

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Related in: MedlinePlus

Color photograph and fluorescein angiography of the iris of monkey number 7 which underwent fake intravitreal injection. Laser irradiation was performed to occlude all major branched retinal veins of the monkey’s eyes (A). Obvious venous dilation and corresponding retinal hemorrhage can be observed from 2 days after laser coagulation (B); at 7 days after laser coagulation, mild iris neovascularization can be observed at the margin of pupil (C) with slight leakage of fluorescein (D); and at 14 days after laser coagulation, severe and tortuous iris neovascularization can be observed across the whole iris (E) with strong leakage of fluorescein (F) which lasted to 23 days after laser coagulation (G and H).
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f3: Color photograph and fluorescein angiography of the iris of monkey number 7 which underwent fake intravitreal injection. Laser irradiation was performed to occlude all major branched retinal veins of the monkey’s eyes (A). Obvious venous dilation and corresponding retinal hemorrhage can be observed from 2 days after laser coagulation (B); at 7 days after laser coagulation, mild iris neovascularization can be observed at the margin of pupil (C) with slight leakage of fluorescein (D); and at 14 days after laser coagulation, severe and tortuous iris neovascularization can be observed across the whole iris (E) with strong leakage of fluorescein (F) which lasted to 23 days after laser coagulation (G and H).

Mentions: Laser irradiation was successfully performed to occlude all major branched retinal veins of the monkeys’ eyes (Figure 3A). In monkey number 7, which served as sham control, obvious venous dilation and corresponding retinal hemorrhage can be observed from 2 days after laser coagulation (Figure 3B); at 7 days after laser coagulation, mild iris neovascularization can be observed at the margin of the pupil with slight leakage of fluorescein (Figure 3C,D); and at 14 days after laser coagulation, severe and tortuous iris neovascularization can be observed at the whole iris with strong leakage of fluorescein (Figure 3E,F), which lasted to 1 month after laser coagulation (Figure 3G,H).


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

Yuan MK, Tao Y, Yu WZ, Kai W, Jiang YR - Mol. Vis. (2010)

Color photograph and fluorescein angiography of the iris of monkey number 7 which underwent fake intravitreal injection. Laser irradiation was performed to occlude all major branched retinal veins of the monkey’s eyes (A). Obvious venous dilation and corresponding retinal hemorrhage can be observed from 2 days after laser coagulation (B); at 7 days after laser coagulation, mild iris neovascularization can be observed at the margin of pupil (C) with slight leakage of fluorescein (D); and at 14 days after laser coagulation, severe and tortuous iris neovascularization can be observed across the whole iris (E) with strong leakage of fluorescein (F) which lasted to 23 days after laser coagulation (G and H).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2927369&req=5

f3: Color photograph and fluorescein angiography of the iris of monkey number 7 which underwent fake intravitreal injection. Laser irradiation was performed to occlude all major branched retinal veins of the monkey’s eyes (A). Obvious venous dilation and corresponding retinal hemorrhage can be observed from 2 days after laser coagulation (B); at 7 days after laser coagulation, mild iris neovascularization can be observed at the margin of pupil (C) with slight leakage of fluorescein (D); and at 14 days after laser coagulation, severe and tortuous iris neovascularization can be observed across the whole iris (E) with strong leakage of fluorescein (F) which lasted to 23 days after laser coagulation (G and H).
Mentions: Laser irradiation was successfully performed to occlude all major branched retinal veins of the monkeys’ eyes (Figure 3A). In monkey number 7, which served as sham control, obvious venous dilation and corresponding retinal hemorrhage can be observed from 2 days after laser coagulation (Figure 3B); at 7 days after laser coagulation, mild iris neovascularization can be observed at the margin of the pupil with slight leakage of fluorescein (Figure 3C,D); and at 14 days after laser coagulation, severe and tortuous iris neovascularization can be observed at the whole iris with strong leakage of fluorescein (Figure 3E,F), which lasted to 1 month after laser coagulation (Figure 3G,H).

Bottom Line: At 50 days after laser treatment, only a slight ectropion uvea was found.However, in the other eyes, obvious INV or hyphema was observed.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, People's Hospital, Peking University, Beijing, China.

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.

Show MeSH
Related in: MedlinePlus