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Lentiviral gene replacement therapy of retinas in a mouse model for Usher syndrome type 1B

Gene Therapy volume 14, pages 584–594 (2007)Cite this article

Abstract

One of the most disabling forms of retinal degeneration occurs in Usher syndrome, since it affects patients who already suffer from deafness. Mutations in the myosin VIIa gene (MYO7A) cause a major subtype of Usher syndrome, type 1B. Owing to the loss of function nature of Usher 1B and the relatively large size of MYO7A, we investigated a lentiviral-based gene replacement therapy in the retinas of MYO7A-null mice. Among the different promoters tested, a CMV-MYO7A chimeric promoter produced wild-type levels of MYO7A in cultured RPE cells and retinas in vivo. Efficacy of the lentiviral therapy was tested by using cell-based assays to analyze the correction of previously defined, MYO7A-null phenotypes in the mouse retina. In vitro, defects in phagosome digestion and melanosome motility were rescued in primary cultures of RPE cells. In vivo, the normal apical location of melanosomes in RPE cells was restored, and the abnormal accumulation of opsin in the photoreceptor connecting cilium was corrected. These results demonstrate that a lentiviral vector can accommodate a large cDNA, such as MYO7A, and mediate correction of important cellular functions in the retina, a major site affected in the Usher syndrome. Therefore, a lentiviral-mediated gene replacement strategy for Usher 1B therapy in the retina appears feasible.

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Acknowledgements

We thank Dr Inder Verma for providing the third generation, self-inactivating lentiviral vector, and Dr Tama Hasson for providing the pM7-10a clone, used in the construction of the MYO7A cDNA. This research was supported by grants from the Foundation Fighting Blindness, Research to Prevent Blindness Foundation, and the NIH (EY14440, EY07042, and core Grants EY12598 and EY00331).

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Author notes

  1. T Hashimoto, D Gibbs and C Lillo: These authors contributed equally to this work.

Authors and Affiliations

  1. Jules Stein Eye Institute, Molecular Biology Institute, UCLA School of Medicine, Los Angeles, CA, USA

    T Hashimoto, X-M Zhang & X-J Yang

  2. Departments of Pharmacology and Neurosciences, UCSD School of Medicine, La Jolla, CA, USA

    D Gibbs, C Lillo, S M Azarian, E Legacki & D S Williams

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  1. T Hashimoto
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  2. D Gibbs
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  3. C Lillo
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Correspondence to X-J Yang or D S Williams.

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

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Hashimoto, T., Gibbs, D., Lillo, C. et al. Lentiviral gene replacement therapy of retinas in a mouse model for Usher syndrome type 1B. Gene Ther 14, 584–594 (2007). https://doi.org/10.1038/sj.gt.3302897

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