May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Biodistribution of rAAV–2/2, rAAV–2/4 and rAAV–2/5 following subretinal or intravitreal administration of rAAV vectors in rat, dog and primate
Author Affiliations & Notes
  • F. Rolling
    Laboratoire de Therapie Genique,
    CHU HOTEL DIEU, Nantes, France
  • N. Provost
    Laboratoire de Therapie Genique,
    CHU HOTEL DIEU, Nantes, France
  • G. Le Meur
    Laboratoire de Therapie Genique,
    CHU HOTEL DIEU, Nantes, France
  • M. Weber
    Service d'ophtalmologie,
    CHU HOTEL DIEU, Nantes, France
  • A. Mendes–Madeira
    Laboratoire de Therapie Genique,
    CHU HOTEL DIEU, Nantes, France
  • G. Podevin
    Service d'ophtalmologie,
    CHU HOTEL DIEU, Nantes, France
  • Y. Cherel
    Laboratoire d'anatomie pathologique, Ecole veterinaire de Nantes, Nantes, France
  • P. Moullier
    Laboratoire de Therapie Genique,
    CHU HOTEL DIEU, Nantes, France
  • Footnotes
    Commercial Relationships  F. Rolling, None; N. Provost, None; G. Le Meur, None; M. Weber, None; A. Mendes–Madeira, None; G. Podevin, None; Y. Cherel, None; P. Moullier, None.
  • Footnotes
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Investigative Ophthalmology & Visual Science May 2004, Vol.45, 3710. doi:
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      F. Rolling, N. Provost, G. Le Meur, M. Weber, A. Mendes–Madeira, G. Podevin, Y. Cherel, P. Moullier; Biodistribution of rAAV–2/2, rAAV–2/4 and rAAV–2/5 following subretinal or intravitreal administration of rAAV vectors in rat, dog and primate . Invest. Ophthalmol. Vis. Sci. 2004;45(13):3710.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose : Studies evaluating the biodistribution of rAAV will define the safe use of this vector. Due to dissemination of rAAV in systemic circulation, distant organs can be exposed to the vector. The purpose of our study was to evaluate the biodistribution of rAAVCMV.gfp vectors following subretinal or intravitreal injection in rat, dog and primate. Methods : Subretinal and intravitreal injections of rAAV–2/2 and chimeric rAAV–2/4 and rAAV–2/5 vectors, carrying a CMV.gfp genome, were performed in the right eye of rat, dog or primate. In animals maintained alive, blood and biopsy samples from submandibular lymphe node, liver and gonads were analyzed. In sacrificed animals, samples from several distant organs were taken. Total DNA from peripheral blood mononuclear cells (PBMC) and few other tissues was extracted and analyzed by PCR using primers located in the gfp DNA sequence. Results : In rAAV–2/2 subretinally injected rats, vector sequences were detected in the optic nerve while for the rats injected in the vitreous, vector sequences were found in the optic nerve and in the brain. Three dogs, each receiving either rAAV–2/2, –2/4 or –2/5, and one primate receiving rAAV–2/4 vector were examined for long term studies. In these animals, rAAV sequences were not detectable in the submandibular lymphe node, nor in the liver and the gonads but were reproducibly found in PBMC. For the subretinally rAAV–2/4 injected primate which was sacrificed at two months post injection, more than 25 different tissue samples were analyzed. rAAV sequences were detectable only in the right optic nerve, in the right submandibular lymphe node and in circulating PBMC. Similar analysis in a rAAV–2/2 intravitreally injected dog showed the presence of rAAV sequences in the optic tract, the optical radiation, the colliculus, the cortex, the cerebellum and the medulla. Conclusions: Following subretinal and intravitreal injections, rAAV sequences were never detected in the liver or in the gonads but were found in PBMC. The most striking result was the detection of rAAV sequences in different regions of the brain after intravitreal injection in dog. This result suggest the ability of rAAV–2 to achieve trans–neuronal transport. This findings raise safety concerns which implies additional studies in large animal models.

Keywords: gene transfer/gene therapy • retina • retina 
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