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Restoration of vision in RPE65-deficient Briard dogs using an AAV serotype 4 vector that specifically targets the retinal pigmented epithelium

Gene Therapy volume 14, pages 292–303 (2007)Cite this article

Abstract

Previous studies have tested gene replacement therapy in RPE65-deficient dogs using recombinant adeno-associated virus 2/2 (rAAV2/2), -2/1 or -2/5 mediated delivery of the RPE65 gene. They all documented restoration of dark- and light-adapted electroretinography responses and improved psychophysical outcomes. Use of a specific RPE65 promoter and a rAAV vector that targets transgene expression specifically to the RPE may, however, provide a safer setting for the long-term therapeutic expression of RPE65. Subretinal injection of rAAV2 pseudotyped with serotype 4 (rAAV2/4) specifically targets the RPE. The purpose of our study was to evaluate a rAAV2/4 vector carrying a human RPE65cDNA driven by a human RPE65 promoter, for the ability to restore vision in RPE65−/− purebred Briard dogs and to assess the safety of gene transfer with respect to retinal morphology and function. rAAV2/4 and rAAV2/2 vectors containing similar human RPE65 promoter and cDNA cassettes were generated and administered subretinally in eight affected dogs, ages 8–30 months (n=6 with rAAV2/4, n=2 with rAAV2/2). Although fluorescein angiography and optical coherence tomography examinations displayed retinal abnormalities in treated retinas, electrophysiological analysis demonstrated that restoration of rod and cone photoreceptor function started as soon as 15 days post-injection, reaching maximal function at 3 months post-injection, and remaining stable thereafter in all animals treated at 8–11 months of age. As assessed by the ability of these animals to avoid obstacles in both dim and normal light, functional vision was restored in the treated eye, whereas the untreated contralateral eye served as an internal control. The dog treated at a later age (30 months) did not recover retinal function or vision, suggesting that there might be a therapeutic window for the successful treatment of RPE65−/− dogs by gene replacement therapy.

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Acknowledgements

We thank Matthew Ellinwood for critical reading and editing. We also thank the Vector Core (www.vectors.nantes.inserm.fr) at the University Hospital of Nantes, supported by the Association Française contre les Myopathies (AFM), INSERM and the Fondation pour la Thérapie Génique en Pays de la Loire. This work was also supported by the French Lions Club and the Lions Clubs International foundation (LCIF). We thank the Association du Berger de Brie and the Fédération des Aveugles et Handicapés Visuels de France. We also thank Graham Holder and Chris Hogg for assistance with the ERG.

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Authors and Affiliations

  1. INSERM U649, CHU Hotel-Dieu, Nantes, Cedex, France

    G Le Meur, K Stieger, M Weber, D Nivard, A Mendes-Madeira, N Provost, P Moullier & F Rolling

  2. CHU Hotel-Dieu, Service d'Ophtalmologie, Nantes, France

    G Le Meur & M Weber

  3. Division of Molecular Therapy, Institute of Ophthalmology, University College of London, London, UK

    A J Smith & R R Ali

  4. Ecole Nationale Vétérinaire de Nantes, Service d'urgences, Nantes, France

    J Y Deschamps

  5. CHU Hotel-Dieu, Laboratoire d'Exploration fonctionnelle, Nantes, France

    Y Péréon

  6. INRA UMR 703, Ecole Nationale Vétérinaire de Nantes, Nantes, France

    Y Cherel

  7. INSERM U583, Hôpital Saint-Eloi, Institut des Neurosciences de Montpellier, Montpellier, France

    C Hamel

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  1. G Le Meur
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Correspondence to F Rolling.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Le Meur, G., Stieger, K., Smith, A. et al. Restoration of vision in RPE65-deficient Briard dogs using an AAV serotype 4 vector that specifically targets the retinal pigmented epithelium. Gene Ther 14, 292–303 (2007). https://doi.org/10.1038/sj.gt.3302861

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