December 2008
Volume 8, Issue 17
Free
OSA Fall Vision Meeting Abstract  |   December 2008
Analysis of a primate cone mosaic following targeted gene transfer using recombinant adeno-associated virus vectors
Author Affiliations
  • Katherine Mancuso
    Katherine Mancuso, Dept. of Ophthalmology, Medical College of Wisconsin, Milwaukee, WI, USA
  • William W. Hauswirth
    WilliamW. Hauswirth, Department of Ophthalmology and Powell Gene Therapy Center, University of Florida, Gainesville, FL, USA
  • Thomas B. Connor
    Thomas B. Connor, Dept. of Ophthalmology, Medical College of Wisconsin, Milwaukee, WI, USA
  • James A. Kuchenbecker
    James A. Kuchenbecker, Dept. of Ophthalmology, Medical College of Wisconsin, Milwaukee, WI, USA
  • Matthew C. Mauck
    Matthew C. Mauck, Dept. of Ophthalmology, Medical College of Wisconsin, Milwaukee, WI, USA
  • Jay Neitz
    Jay Neitz, Dept. of Ophthalmology, Medical College of Wisconsin, Milwaukee, WI, USA
  • Maureen Neitz
    Maureen Neitz, Dept. of Ophthalmology, Medical College of Wisconsin, Milwaukee, WI, USA
Journal of Vision December 2008, Vol.8, 69. doi:10.1167/8.17.69
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      Katherine Mancuso, William W. Hauswirth, Thomas B. Connor, James A. Kuchenbecker, Matthew C. Mauck, Jay Neitz, Maureen Neitz; Analysis of a primate cone mosaic following targeted gene transfer using recombinant adeno-associated virus vectors. Journal of Vision 2008;8(17):69. doi: 10.1167/8.17.69.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

We have successfully used gene therapy to add red-green color vision, as assessed in behavioral tests, in dichromatic squirrel monkeys that had only short- (S) and middle- (M) wavelength-sensitive cones. Expression of a human long- (L) wavelength sensitive opsin transgene was targeted to the primate M-cones through subretinal injections of a recombinant adeno-associated virus vector containing the L/M opsin enhancer and promoter. By coexpressing the L-opsin transgene in a subset of the endogenous M-cones, the treatment shifted the spectral sensitivity of the transduced M-cones to respond to long wavelength light, thus producing a retina with two distinct cone types absorbing in the middle-to-long wavelengths, as required for trichromacy. A remaining question is how well does the cone mosaic produced by gene therapy mimic the normal human L/M cone mosaic? To answer this question, a squirrel monkey was injected with a 2:1 mixture of L-opsin-coding virus and an identical virus containing a green fluorescent protein (GFP) gene. Color multifocal electroretinogram analysis under conditions that strongly favor responses from the introduced L-opsin showed elevated amplitudes in discrete areas that corresponded to locations where subretinal injections were placed, as confirmed by in vivo fluorescence imaging of GFP transgene expression. Confocal microscopy, performed approximately 2 years and 3 months post-injection, revealed a mosaic pattern of GFP transgene expression in 5–12% of the cones in treated areas. Because the GFP-coding virus was diluted to 1/3 compared to the opsin-coding virus used in the behaviorally tested monkeys, we estimate that 15–36% of cones in those animals are transduced.

Mancuso, K. Hauswirth, W. W. Connor, T. B. Kuchenbecker, J. A. Mauck, M. C. Neitz, J. Neitz, M. (2008). Analysis of a primate cone mosaic following targeted gene transfer using recombinant adeno-associated virus vectors [Abstract]. Journal of Vision, 8(17):69, 69a, http://journalofvision.org/8/17/69/, doi:10.1167/8.17.69. [CrossRef]
Footnotes
 Supported by the National Institutes of Health grants R03EY014056, R01EY016861, R01EY11123, Research Training Program in Vision Science Grant T32EY014537, NEI Core Grant for Vision Research EY01981, the Harry J. Heeb Foundation, and Research to Prevent Blindness.
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