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S. Kachi, K. Binley, N. Umeda, H. Akiyama, K. Yokoi, M. Kachi, W. Xiao, M. Espa, S. Naylor, P.A. Campochiaro; Delivery of Angioinhibitory Genes Using the VMD2 Promoter in Lentiviral Vectors Results in Selective Expression in RPE and Inhibition of Choroidal Neovascularization . Invest. Ophthalmol. Vis. Sci. 2006;47(13):906.
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© ARVO (1962-2015); The Authors (2016-present)
In this study, we investigated the potential for using the Equine Infectious Anemia Virus (EIAV) Vectors for treatment of choroidal neovascularization (CNV).
Adult Balb/C mice were given subretinal injection of EIAV–CMV–LacZ or EIAV–VMD2–LacZ. LacZ was localized by histochemistry and quantified using ß–gal ELISA. Adult C57BL/6 mice were given subretinal injection of one of following: EIAV–Null, EIAV–CMV–hEndostatin, EIAV–CMV–mEndostatin, EIAV–VMD2–mEndostatin. On day 14 after injection, Bruch’s membrane was ruptured with laser photocoagulation and after 14 days, mice were perfused with fluorescein–labeled dextran and choroidal flat mounts were made to measure the CNV size.
The CMV promoter drove strong transgene expression predominantly in the RPE, but there was also some expression in the optic nerve, while the VMD2 promoter caused selective expression in the RPE. Quantitation of the whole eye revealed VMD2 mediated expression was reduced 6–10 fold cmpared to CMV. Expression occurred for at least 8 weeks, the longest time point examined. Expression of hEndostatin had no significant effect on CNV size, while compared to injection of EIAV–null, expression of mEndostatin with the CMV or VMD2 promoter reduced CNV size by 60% or 35%, respectively.
These data suggest that long–term expression of transgenes can be achieved after subretinal injection of EIAV vectors using either a CMV or VMD2 promoter; expression driven by the VMD2 promoter is completely RPE–specific, but is at a somewhat lower level. Expression of murine, but not human, endostatin with either promoter inhibited CNV. This supports the feasibility of EIAV vectors with either a CMV or VMD2 promoter depending upon the potency of the transgene and the level of concern regarding ectopic expression with the CMV promoter. This study also suggests the potential importance of using transgenes derived from the species of the host.
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