April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
rAAV-mediated Gene Delivery of Truncated Canine RPGR Causes Photoreceptor Dysplasia in Dogs with RPGR-ORF15 Stop Mutation
Author Affiliations & Notes
  • William A. Beltran
    Clinical Studies, Univ of Pennsylvania Sch Vet Med, Philadelphia, Pennsylvania
  • Simone Iwabe
    Clinical Studies, Univ of Pennsylvania Sch Vet Med, Philadelphia, Pennsylvania
  • Diego S. Fajardo
    Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida
  • Sanford Boye
    Dept of Ophthalmology, Univ. of Florida, Gainesville, Florida
  • Wen-Tao Deng
    Dept of Ophthalmology, Univ. of Florida, Gainesville, Florida
  • William W. Hauswirth
    Dept of Ophthalmology, Univ. of Florida, Gainesville, Florida
  • Alfred S. Lewin
    Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida
  • Gustavo D. Aguirre
    Clinical Studies, Univ of Pennsylvania Sch Vet Med, Philadelphia, Pennsylvania
  • Footnotes
    Commercial Relationships  William A. Beltran, None; Simone Iwabe, None; Diego S. Fajardo, None; Sanford Boye, None; Wen-Tao Deng, None; William W. Hauswirth, AGTC (P); Alfred S. Lewin, None; Gustavo D. Aguirre, None
  • Footnotes
    Support  EY13132, EY06855, EY17549, EY11123, FFB, MVRF, Fight for Sight, RPB Inc.
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 1418. doi:
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      William A. Beltran, Simone Iwabe, Diego S. Fajardo, Sanford Boye, Wen-Tao Deng, William W. Hauswirth, Alfred S. Lewin, Gustavo D. Aguirre; rAAV-mediated Gene Delivery of Truncated Canine RPGR Causes Photoreceptor Dysplasia in Dogs with RPGR-ORF15 Stop Mutation. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1418.

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Abstract

Purpose: : To assess the efficiency and safety of RPGR replacement constructs delivered with a recombinant adeno-associated viral serotype 5 vector (rAAV2/5) in XLPRA1 dogs that carry a stop mutation in RPGR exon ORF15.

Methods: : Three different promoters that included either segment of the proximal mouse opsin promoter (mOp), the human G-protein coupled receptor protein kinase 1 promoter (hGRK1), or the human interphotoreceptor retinoid binding protein promoter (hIRBP) were used to drive the expression of either the full-length human RPGR-ORF15 cDNA (hRPGR), or a shortened version of the canine RPGR cDNA (cRPGRshort). Four constructs (mOP-cRPGRshort, hIRBP-cRPGRshort, hGRK1-hRPGR, and hIRBP-hRPGR) were generated, packaged in rAAV2/5 vectors, and each was delivered via subretinal injection (150 ul at 1.51 x 1011 vg/ml) to the right eye of 2 mutant dogs at ~ 26-28 wks of age. Left eyes were sham-injected with BSS. Regular ocular examinations, and in vivo imaging (Spectralis HRA/OCT) were performed.

Results: : At 7 wks post-injection multifocal hyper-reflective lesions were observed by indirect ophthalmoscopy and cSLO/OCT imaging in the injected retinal area of the 2 dogs treated with hIRBP-cRPGRshort, and of 1 dog treated with mOP-cRPGRshort. These animals were euthanized, and histologic examination of the retinas showed severe multifocal retinal dysplastic lesions. A fourth dog treated with hGRK1-hRPGR had multifocal retinal detachments that were confirmed by histology. All 4 remaining animals did not show any adverse effects, and are still currently enrolled in the study.

Conclusions: : Multifocal retinal dysplastic lesions were found following subretinal injection of an AAV2/5 vector to deliver a shortened canine RPGR-ORF15 cDNA in rods. Over-expression of the RPGR-ORF15 transgene may be responsible for this loss of normal retinal lamination in some treated animals.

Keywords: gene transfer/gene therapy • retinal degenerations: hereditary • photoreceptors 
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