Purchase this article with an account.
F. Rolling, M. Weber, N. Provost, H. Conrath, S. Folliot, D. Briot, Y. Cherel, J. Rabinowitz, J. Samulski, P. Moullier; Following Subretinal Injection, Recombinant Adeno-Associated Virus (rAAV) Serotype 4 Specifically Transduces Retinal Pigmented Epithelium in Rat, Dog and Macaque . Invest. Ophthalmol. Vis. Sci. 2003;44(13):446.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To evaluate gene transfer efficiency and tropism of rAAV serotypes -4 and -5 following subretinal injection in rat, dog and nonhuman primate. Vector biodistribution was also examined. Methods: Subretinal injections of rAAV-4 and -5 carrying an AAV-2-CMV.gfp genome were performed in Wistar rats, Beagle dogs and macaques. Green fluorescent protein (GFP) protein expression in live animals was monitored by fluorescent retinal imaging. Viral tropism was determined by histology. Vector distribution was monitored by PCR in various biological fluids for one month post rAAV administration. Results: Serotypes -4 and –5 displayed stable GFP expression over a six months period in dogs (duration of the experiment). Retinal flat-mounts were performed in order to analyse the transduction pattern. Similarly to AAV-2, AAV-5 transduced both RPE and photoreceptor cells, with higher level of transduction in photoreceptors, whereas AAV-4-mediated transduction was restricted to RPE cells. In order to check if the RPE restricted tropism of AAV-4 was conserved across species, subretinal injection of AAV-4 was performed in nonhuman primate. Stable GFP expression was detected in macaques retina for a three months period (duration of the experiment) and retina histological sections displayed specific GFP expression in RPE cells. Following rAAV-4 and -5 subretinal delivery in dogs (n=6) and in primates (n=2), vector genome was found in lacrymal and nasal fluids for up to 3 days and in the serum for 15-20 days. Conclusions: RPE specific transduction in rat, dog and primate, makes AAV-4-derived vectors attractive for retinal disease originating in RPE such as Leber Congenital Amaurosis (RPE65). Similarly to AAV-2, AAV-5 displayed an efficient transduction of photoreceptors as well. Vector biodistribution in biological fluids of 8 large animals displayed an identical pattern.
This PDF is available to Subscribers Only