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Retinal gene therapy with a large MYO7A cDNA using adeno-associated virus

Gene Therapy volume 20, pages 824–833 (2013)Cite this article

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Abstract

Usher 1 patients are born profoundly deaf and then develop retinal degeneration. Thus they are readily identified before the onset of retinal degeneration, making gene therapy a viable strategy to prevent their blindness. Here, we have investigated the use of adeno-associated viruses (AAVs) for the delivery of the Usher 1B gene, MYO7A, to retinal cells in cell culture and in Myo7a-null mice. MYO7A cDNA, under control of a smCBA promoter, was packaged in single AAV2 and AAV5 vectors and as two overlapping halves in dual AAV2 vectors. The 7.9-kb smCBA-MYO7A exceeds the capacity of an AAV vector; packaging of such oversized constructs into single AAV vectors may involve fragmentation of the gene. Nevertheless, the AAV2 and AAV5 single vector preparations successfully transduced photoreceptor and retinal pigment epithelium cells, resulting in functional, full-length MYO7A protein and correction of mutant phenotypes, suggesting successful homologous recombination of gene fragments. With discrete, conventional-sized dual AAV2 vectors, full-length MYO7A was detected, but the level of protein expression was variable, and only a minority of cells showed phenotype correction. Our results show that MYO7A therapy with AAV2 or AAV5 single vectors is efficacious; however, the dual AAV2 approach proved to be less effective.

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Acknowledgements

We thank Nuno Martins for technical assistance. The study was supported by grants from the Foundation Fighting Blindness, the Macula Vision Research Foundation, Research to Prevent Blindness (RPB) and the NIH (EY07042 to DSW, core Grants EY00331 and EY021721 to WWH). CML was supported by NIH training Grant GM008243. HF was supported by an FCT PhD fellowship. DSW is a Jules and Doris Stein RPB Professor.

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  1. V S Lopes and S E Boye: These authors contributed equally to this work.

Authors and Affiliations

  1. Departments of Ophthalmology and Neurobiology, Jules Stein Eye Institute, UCLA School of Medicine, Los Angeles, CA, USA

    V S Lopes, C M Louie & D S Williams

  2. Centre of Ophthalmology, IBILI, Faculty of Medicine, University Coimbra, Coimbra, Portugal

    V S Lopes & H Fofo

  3. Department of Ophthalmology, College of Medicine, University of Florida, Gainsville, FL, USA

    S E Boye, S Boye, F Dyka, V Chiodo & W W Hauswirth

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  1. V S Lopes
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Correspondence to D S Williams.

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WWH and the University of Florida have a financial interest in the use of AAV therapies and own equity in a company (AGTC Inc.), which might, in the future, commercialize some aspects of this work.

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Lopes, V., Boye, S., Louie, C. et al. Retinal gene therapy with a large MYO7A cDNA using adeno-associated virus. Gene Ther 20, 824–833 (2013). https://doi.org/10.1038/gt.2013.3

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