May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Following Subretinal Injection, Recombinant Adeno-Associated Virus (rAAV) Serotype 4 Specifically Transduces Retinal Pigmented Epithelium in Rat, Dog and Macaque
Author Affiliations & Notes
  • F. Rolling
    Laboratoire de Therapie Genique, CHU HOTEL DIEU, Nantes, France
  • M. Weber
    Service d'ophtalmologie, CHU HOTEL DIEU, Nantes, France
  • N. Provost
    Service d'ophtalmologie, CHU HOTEL DIEU, Nantes, France
  • H. Conrath
    Service d'ophtalmologie, CHU HOTEL DIEU, Nantes, France
  • S. Folliot
    Service d'ophtalmologie, CHU HOTEL DIEU, Nantes, France
  • D. Briot
    Service d'ophtalmologie, CHU HOTEL DIEU, Nantes, France
  • Y. Cherel
    Laboratoire d'anatomie pathologique, Ecole veterinaire de Nantes, Nantes, France
  • J. Rabinowitz
    Gene Therapy Center, University of North Carolina, Chape Hill, NC, United States
  • J. Samulski
    Gene Therapy Center, University of North Carolina, Chape Hill, NC, United States
  • P. Moullier
    Gene Therapy Center, University of North Carolina, Chape Hill, NC, United States
  • Footnotes
    Commercial Relationships  F. Rolling, None; M. Weber, None; N. Provost, None; H. Conrath, None; S. Folliot, None; D. Briot, None; Y. Cherel, None; J. Rabinowitz, None; J. Samulski, None; P. Moullier, None.
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 446. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      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)

      ×
  • Supplements
Abstract

Abstract: : 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.

Keywords: gene transfer/gene therapy • retinal pigment epithelium 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×