Adeno-Associated Virus Gene Transfer to Mouse Retina

Abstract

Ocular gene transfer may provide a means for arresting the retinal degeneration characteristic of many inherited causes of blindness, including retinitis pigmentosa (RP). Previously, we have shown in immunodeficient animals that recombinant adeno-associated virus (rAAV) mediates transduction of photoreceptors as well as the retinal pigment epithelium (RPE) following subretinal injection. In this study we extend these observations and show that highly purified recombinant AAV vectors encoding the reporter gene LacZ transduce photoreceptors in an immunocompetent mouse strain following subretinal injection and efficiently transduce ganglion cells after intravitreal injection. Levels of transduction increase over time. Sublethal gamma-irradiation is shown to facilitate this process.

Novel anti-VEGF chimeric molecules delivered by AAV vectors for inhibition of retinal neovascularization

“…25,26 There are several routes to deliver gene therapy vectors into the eye, however the most common ones are intravitreal and subretinal. 19,21 We first decided to investigate an intravitreal route of delivery because of its potential ease of use in the retinal clinic. The AAV2.sFLT01 vector predominantly transduced retinal ganglion cells in agreement with previous observation by other investigators.…”

“…The AAV2.sFLT01 vector predominantly transduced retinal ganglion cells in agreement with previous observation by other investigators. 21 Subretinal delivery of AAV2 encoding full-length sFlt-1 gene has been successfully tested in mouse and in primate models of ocular neovascularization. 22,27,28 An adenoviral vector encoding full-length sFlt-1, injected both intravitreously or periocularly suppressed choroidal neovascularization at rupture sites in Bruch's membrane.…”

“…18 Adeno-associated virus (AAV) vectors offer an attractive tool for intraocular gene delivery because of their nonpathogenic nature, low toxicity, minimal immunogenicity and long-term persistence. [19][20][21][22] Intravitreal administration of AAV serotype 2 (AAV2) vector in mice results mostly in transduction of ganglion cells and few cells in the inner nuclear layer. 21 Subretinal delivery of AAV2 vector encoding full-length sFlt-1 (transduction of photoreceptors and retinal pigmented epithelium) prevented development of laser-induced choroidal neovascularization in all treated monkeys.…”

“…[19][20][21][22] Intravitreal administration of AAV serotype 2 (AAV2) vector in mice results mostly in transduction of ganglion cells and few cells in the inner nuclear layer. 21 Subretinal delivery of AAV2 vector encoding full-length sFlt-1 (transduction of photoreceptors and retinal pigmented epithelium) prevented development of laser-induced choroidal neovascularization in all treated monkeys. 22 In this study we have designed and constructed small novel soluble hybrid molecules that have strong anti-VEGF activity in vitro, incorporated these molecules into AAV2 vectors and then tested in vivo persistence of expression and efficacy and safety following intravitreal delivery of these AAV2-based vectors in the oxygeninduced retinopathy of prematurity (OIR) mouse model.…”

Recombinant AAV-mediated gene transfer to the retina: gene therapy perspectives

“…[9][10][11][12][13][14][15][16] In the retina, following subretinal delivery, AAV-2 vectors transduce RPE and photoreceptor cells. [17][18][19] Intravitreal injection of rAAV-2 leads to efficient ganglion cell transduction. [20][21][22][23] We and others have studied rAAV chimeric serotypes in which the vector is flanked by AAV-2 ITRs, but encapsidated in an AAV-1, -2, -3, -4 or -5 shell generating rAAV-2/1,-2/2, -2/3, -2/4 and -2/5, respectively.…”