Abstract
To test whether fast-acting, self-complimentary (sc), adeno-associated virus-mediated RPE65 expression prevents cone degeneration and/or restores cone function, we studied two mouse lines: the Rpe65-deficient rd12 mouse and the Rpe65-deficient, rhodopsin null (‘that is, cone function-only’) Rpe65−/−::Rho−/− mouse. scAAV5 expressing RPE65 was injected subretinally into one eye of rd12 and Rpe65−/−::Rho−/− mice at postnatal day 14 (P14). Contralateral rd12 eyes were injected later, at P35. Rd12 behavioral testing revealed that rod vision loss was prevented with either P14 or P35 treatment, whereas cone vision was only detected after P14 treatment. Consistent with this observation, P35 treatment only restored rod electroretinogram (ERG) signals, a result likely due to reduced cone densities at this time point. For Rpe65−/−::Rho−/− mice in which there is no confounding rod contribution to the ERG signal, cone cells and cone-mediated ERGs were also maintained with treatment at P14. This work establishes that a self-complimentary AAV5 vector can restore substantial visual function in two genetically distinct models of Rpe65 deficiency within 4 days of treatment. In addition, this therapy prevents cone degeneration but only if administered before extensive cone degeneration, thus supporting continuation of current Leber's congenital amaurosis-2 clinical trials with an added emphasis on cone subtype analysis and early intervention.
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Acknowledgements
We thank Vince Chiodo and Thomas Doyle at the University of Florida and Keqing Zhang at the University of Missouri for the technical support. We also acknowledge NIH Grants EY018331, EY13729, EY11123, NS36302, EY08571, EY07758, EY014046, EY06360, EY017246 (to DE), EY00067 (to RB) and grants from the Macular Vision Research Foundation, Foundation Fighting Blindness, Fight for Sight (to DE), Lions of Central NY, NASA (to RB), Juvenile Diabetes Research Foundation and Research to Prevent Blindness Inc. for partial support of this work.
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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.) that might, in the future, commercialize some aspects of this work. There is no conflict of interest for other co-authors.
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Pang, J., Boye, S., Lei, B. et al. Self-complementary AAV-mediated gene therapy restores cone function and prevents cone degeneration in two models of Rpe65 deficiency. Gene Ther 17, 815–826 (2010). https://doi.org/10.1038/gt.2010.29
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DOI: https://doi.org/10.1038/gt.2010.29
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