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Single residue AAV capsid mutation improves transduction of photoreceptors in the Abca4−/− mouse and bipolar cells in the rd1 mouse and human retina ex vivo

Gene Therapy volume 23, pages 767–774 (2016)Cite this article

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Abstract

Gene therapy using adeno-associated viral (AAV) vectors for the treatment of retinal degenerations has shown safety and efficacy in clinical trials. However, very high levels of vector expression may be necessary for the treatment of conditions such as Stargardt disease where a dual vector approach is potentially needed, or in optogenetic strategies for end-stage degeneration in order to achieve maximal light sensitivity. In this study, we assessed two vectors with single capsid mutations, rAAV2/2(Y444F) and rAAV2/8(Y733F) in their ability to transduce retina in the Abca4−/− and rd1 mouse models of retinal degeneration. We noted significantly increased photoreceptor transduction using rAAV2/8(Y733F) in the Abca4−/− mouse, in contrast to previous work where vectors tested in this model have shown low levels of photoreceptor transduction. Bipolar cell transduction was achieved following subretinal delivery of both vectors in the rd1 mouse, and via intravitreal delivery of rAAV2/2(Y444F). The successful use of rAAV2/8(Y733F) to target bipolar cells was further validated on human tissue using an ex vivo culture system of retinal explants. Capsid mutant AAV vectors transduce human retinal cells and may be particularly suited to treat retinal degenerations in which high levels of transgene expression are required.

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Acknowledgements

This study was supported by Wellcome Trust, NIHR Biomedical Research Centres of Oxford and Moorfields, MRC, Fight for Sight, Royal College of Surgeons ProRetina.

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Author notes

  1. S R De Silva and P Charbel Issa: These authors contributed equally to this work.

Authors and Affiliations

  1. Nuffield Laboratory of Ophthalmology, NIHR Biomedical Research Centre, University of Oxford, Oxford, UK

    S R De Silva, P Charbel Issa, M S Singh, D M Lipinski, A O Barnea-Cramer, N J Walker, A R Barnard, M W Hankins & R E MacLaren

  2. Department of Ophthalmology, University of Bonn, Bonn, Germany

    P Charbel Issa

  3. Moorfields Eye Hospital, NIHR Biomedical Research Centre, Oxford, UK

    R E MacLaren

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  1. S R De Silva
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  2. P Charbel Issa
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  3. M S Singh
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  4. D M Lipinski
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Correspondence to M W Hankins or R E MacLaren.

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Competing interests

REM is a Founder and Director of Nightstarx Ltd (Gibbs Building, 215 Euston Road, London NW1 2BE), a choroideremia gene therapy company established by the University of Oxford and funded by the Wellcome Trust through Syncona Partners.

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De Silva, S., Charbel Issa, P., Singh, M. et al. Single residue AAV capsid mutation improves transduction of photoreceptors in the Abca4−/− mouse and bipolar cells in the rd1 mouse and human retina ex vivo. Gene Ther 23, 767–774 (2016). https://doi.org/10.1038/gt.2016.54

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