Stable RAAV-Mediated Transduction of Rod and Cone Photoreceptors in the Canine Retina
Abstract
Recombinant adeno-associated virus (rAAV) vectors are attractive candidates for the treatment of inherited and acquired retinal disease. Although rAAV vectors are well characterized in rodent models, a prerequisite to their clinical application in human patients is the thorough evaluation of their efficacy and safety in intermediate animal models. In this study, we describe rAAV-2-mediated expression of GFP reporter gene in retinal cells following local vector delivery in dogs. Subretinal delivery of rAAV.CMV.GFP was performed unilaterally in eight normal dogs from 6 weeks of age. The area of retinal transduction was maximized by the optimization of surgical techniques for subretinal vector delivery by pars-plana vitrectomy and the use of fine-gauge subretinal cannulae to create multiple retinotomies. rAAV-2 vectors mediated efficient stable reporter gene expression in photoreceptors and retinal pigment epithelial cells. We found efficient transduction of cone photoreceptors in addition to rods in both the canine retina and after subretinal vector delivery in another intermediate animal model, the feline retina. GFP expression in dogs was confined to the area of the retinal bleb and was sustained in cells at this site for at least 18 months. Electroretinography demonstrated a modest reduction in global rod-mediated retinal function following subretinal delivery of rAAV.CMV.GFP. Three of the eight animals developed delayed-onset intraocular inflammation, in two cases associated with a serum antibody response to GFP protein. We conclude that rAAV-2 vectors mediate efficient sustained transgene expression in rod and cone photoreceptors following subretinal delivery in this intermediate animal model. The possibility of adverse effects including intraocular immune responses and reduced retinal function requires further investigation prior to clinical applications in patients.
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In canines, rAAV2 carrying GFP delivered to the retina by subretinal injection produced long-term expression in photoreceptors from 2 weeks to 12 months, but evidence of intraocular inflammatory response was reported in three out of eight animals.
“In canines, a study designed to evaluate the efficiency and safety of AAV vectors, delivered rAAV2 carrying GFP to the retina by subretinal injection.Long-term expression was observed in photoreceptors starting from 2 weeks and persisting until 12 months.However, evidence of an intraocular inflammatory response was reported in three out of eight animals [49].”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [49]
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- Authors
- Bainbridge, J. W. B., Mistry, A., Schlichtenbrede, F. C., Smith, A., Broderick, C., De Alwis, M., Georgiadis, A., Taylor, P. M., Squires, M., Sethi, C., Charteris, D., Thrasher, A. J., Sargan, D., & Ali, R. R.
- Affiliations
- University College London (GB)
- University of Cambridge (GB)
- ORCID
- Venue
- Gene Therapy
- Published
- 2003-08-01
- Volume
- 10
- Issue
- 16
- Pages
- 1336-1344
- ISSN
- 0969-7128 (ISSN-L)
- 1476-5462
- OpenAlex
- W2025109749
- Cited by
- 58
- References
- 25
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 2 · 2023: 1 · 2022: 1 · 2021: 1 · 2020: 1 · 2019: 1 · 2018: 2 · 2017: 4 · 2016: 1 · 2014: 3 · 2013: 3 · 2012: 3
- FWCI
- 0.92
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Gene Therapy (publishedVersion)
- PubMed (publishedVersion)
- UCL Discovery (University College London) (submittedVersion)
- Funders
- Wellcome Trust
- Sir Jules Thorn Charitable Trust
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- recombinant adeno-associated virus, subretinal gene delivery, photoreceptors, rod photoreceptors, cone photoreceptors, retinal pigment epithelium, canine retina, pars plana vitrectomy, retinotomy, GFP reporter gene, sustained transgene expression, intraocular inflammation, antibody response, electroretinogram, retinal gene therapy
- MeSH terms
- Dependovirus, Animals, Cells, Cultured, Dogs, Electroretinography, Fundus Oculi, Genetic Vectors, Inflammation, Luminescent Proteins, Pigment Epithelium of Eye, Retinal Diseases, Time Factors, Transduction, Genetic, Genetic Therapy, Gene Expression, Photoreceptor Cells, Vertebrate, Green Fluorescent Proteins
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