title: Lentivirus-mediated RNA interference of vascular endothelial growth factor in monkey eyes with iris neovascularization.
abstract: OBJECTIVE
  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.
authors.affiliation: Department of Ophthalmology, People's Hospital, Peking University, Beijing, China.
authors.forename: Meng-Ke
authors.initials: MK
authors.lastname: Yuan
authors.forename: Yong
authors.initials: Y
authors.lastname: Tao
authors.forename: Wen-Zhen
authors.initials: WZ
authors.lastname: Yu
authors.forename: Wang
authors.initials: W
authors.lastname: Kai
authors.forename: Yan-Rong
authors.initials: YR
authors.lastname: Jiang
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005451:Fluorescein Angiography; D049452:Green Fluorescent Proteins / Q000378:metabolism; D000882:Haplorhini; D007150:Immunohistochemistry; D007498:Iris / Q000098:blood supply* / Q000473:pathology*; D016086:Lentivirus / Q000378:metabolism*; D009389:Neovascularization, Pathologic / Q000175:diagnosis / Q000628:therapy*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A; D049452:Green Fluorescent Proteins
references.citation: Breast Cancer Res Treat. 1995;36(2):169-80
references.pmid: 8534865
references.citation: Semin Oncol. 2002 Dec;29(6 Suppl 16):10-4
references.pmid: 12516033
references.citation: Chin Med J (Engl). 2008 Mar 5;121(5):439-44
references.pmid: 18364118
references.citation: Neoplasia. 2008 Apr;10(4):399-407
references.pmid: 18392139
references.citation: Doc Ophthalmol. 1976 Jan 28;40(1):1-241
references.pmid: 765083
references.citation: J Surg Res. 2004 Apr;117(2):339-44
references.pmid: 15047140
references.citation: Nature. 1998 Feb 19;391(6669):806-11
references.pmid: 9486653
references.citation: Retina. 2005 Feb-Mar;25(2):193-201
references.pmid: 15689811
references.citation: Gene Ther. 2005 Apr;12(8):694-701
references.pmid: 15660111
references.citation: Gene Ther. 2006 Feb;13(3):225-34
references.pmid: 16195704
references.citation: Blood. 2000 Nov 15;96(10):3392-8
references.pmid: 11071633
references.citation: Am J Pathol. 1994 Sep;145(3):574-84
references.pmid: 7521577
references.citation: N Engl J Med. 1994 Dec 1;331(22):1480-7
references.pmid: 7526212
references.citation: Ophthalmology. 2001 Oct;108(10):1767-76; quiz1777, 1800
references.pmid: 11581047
references.citation: Ophthalmology. 1998 Feb;105(2):232-7
references.pmid: 9479280
references.citation: Ophthalmology. 2008 Sep;115(9):1571-80, 1580.e1-3
references.pmid: 18440643
references.citation: Adv Exp Med Biol. 2006;572:195-200
references.pmid: 17249575
references.citation: Blood. 2004 Jun 15;103(12):4511-3
references.pmid: 14988151
references.citation: Mol Vis. 2003 May 30;9:210-6
references.pmid: 12789138
references.citation: Acta Ophthalmol. 2009 Nov;87(7):736-40
references.pmid: 18803622
references.citation: Rev Med Virol. 2003 Nov-Dec;13(6):373-85
references.pmid: 14625885
references.citation: Zhonghua Yan Ke Za Zhi. 2009 Aug;45(8):746-51
references.pmid: 20021890
references.citation: Arch Dermatol Res. 2008 Apr;300(4):177-84
references.pmid: 18239926
references.citation: Invest Ophthalmol Vis Sci. 2004 May;45(5):1289-96
references.pmid: 15111579
references.citation: Arch Ophthalmol. 1989 Apr;107(4):581-6
references.pmid: 2468330
references.citation: Curr Eye Res. 2006 Feb;31(2):171-80
references.pmid: 16500768
references.citation: Eur J Ophthalmol. 2008 Mar-Apr;18(2):255-62
references.pmid: 18320519
references.citation: Anat Rec (Hoboken). 2009 May;292(5):633-9
references.pmid: 19382240