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Proteosomal degradation impairs transcytosis of AAV vectors from suprachoroidal space to retina

Gene Therapy volume 28, pages 740–747 (2021)Cite this article

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Abstract

Suprachoroidal injection provides a new route of delivery for AAV vectors to retinal pigmented epithelial cells and photoreceptors that can be done in an outpatient setting and is less invasive and potentially safer than subretinal injection, the most common route of delivery for ocular gene therapy. After suprachoroidal injection of AAV8 or AAV9 vectors, there is strong transduction of photoreceptors, but it is unclear how vector traverses the retinal pigmented epithelium. In this study, we found that transduction of photoreceptors was significantly increased after suprachoroidal injection of AAV2tYF-CBA-GFP versus AAV2-CBA-GFP vector. Compared with AAV2, AAV2tYF is more resistant to proteosomal degradation. Treatment with protease inhibitors significantly increased photoreceptor transduction after suprachoroidal injection of AAV5-GRK1-GFP. These data suggest that after suprachoroidal injection, AAV vectors access photoreceptors by transcytosis through retinal pigmented epithelial cells during which they are subject to proteosomal degradation, which if suppressed can enhance transduction of photoreceptors.

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Fig. 1: Good expression of GFP in retina and retinal pigmented epithelium after suprachoroidal injection of AAV2tYF-CBA-GFP but not AAV2-CBA-GFP.
Fig. 2: Strong, selective expression of GFP in photoreceptors after suprachoroidal injection of AAV2tYF-GRK1-GFP.
Fig. 3: Protease inhibitors increase GFP expression in photoreceptors after suprachoroidal injection of AAV5-GRK1-GFP.

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Acknowledgements

The authors thank Applied Genetic Technologies Corporation (Gainesville, FL) for providing AAV vectors for this study.

Funding

Supported by EY031097 from the National Eye Institute and an unrestricted grant from Research to Prevent Blindness, New York, NY.

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Authors and Affiliations

  1. The Wilmer Eye Institute, Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Kun Ding, Jikui Shen, Sean Hackett, Mahmood Khan & Peter A. Campochiaro

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  1. Kun Ding
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  2. Jikui Shen
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  3. Sean Hackett
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Correspondence to Peter A. Campochiaro.

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Conflict of interest

None of the authors have a conflict of interest directly related to the content of the manuscript, but PAC has unrelated financial disclosures: AERPIO PHARMACEUTICALS: Advisory Board, Honoraria to JHU, Investigator, Grant; ALLEGRO: Advisory Board, Equity; APPLIED GENETIC TECHNOLOGIES CORPORATION: Advisory Board, Honoraria;ASCLEPIX THERAPEUTICS: Consultant, Honoraria; BAUCSH and LOMB: Consultant, Honoraria; CUREVAC: Consultant, Honoraria; EXONATE LTD: Advisory Board, Honoraria GENENTECH/ROCHE INC: Advisory Board, Honoraria to JHU, Investigator, Grants; SANOFI GENZYME: Investigator, Grant; GRAYBUG VISION: Consultant, Honoraria, Co-Founder, Equity, Grant; MERCK & CO, INC: Advisory Board, Honoraria; OXFORD BIOMEDICA: Investigator, Grant; PERFUSE; REGENERON PHARMACEUTICALS, INC: Investigator, Grant; REGENXBIO, INC: Investigator, Grant; WAVE LIFE SCIENCES: Consultant, Honoraria.

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Ding, K., Shen, J., Hackett, S. et al. Proteosomal degradation impairs transcytosis of AAV vectors from suprachoroidal space to retina. Gene Ther 28, 740–747 (2021). https://doi.org/10.1038/s41434-021-00233-1

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