Recombinant adeno-associated virus serotype 4 mediates unique and exclusive long-term transduction of retinal pigmented epithelium in rat, dog, and nonhuman primate after subretinal delivery
Open accessAbstract
We previously described chimeric recombinant adeno-associated virus (rAAV) vectors 2/4 and 2/5 as the most efficient vectors in rat retina. We now characterize these two vectors carrying the CMV.gfp genome following subretinal injection in the Wistar rat, beagle dog, and cynomolgus macaque. Both serotypes displayed stable GFP expression for the duration of the experiment (6 months) in all three animal models. Similar to the AAV-2 serotype, AAV-2/5 transduced both RPE and photoreceptor cells, with higher level of transduction in photoreceptors, whereas rAAV-2/4 transduction was unambiguously restricted to RPE cells. This unique specificity found conserved among all three species makes AAV-2/4-derived vectors attractive for retinal diseases originating in RPE such as Leber congenital amaurosis (RPE65) or retinitis pigmentosa due to a mutated mertk gene. To provide further important preclinical data, vector shedding was monitored by PCR in various biological fluids for 2 months post-rAAV administration. Following rAAV-2/4 and -5 subretinal delivery in dogs (n = 6) and in nonhuman primates (n = 2), vector genome was found in lacrymal and nasal fluids for up to 3-4 days and in the serum for up to 15-20 days. Overall, these findings will have a practical impact on the development of future gene therapy trials of retinal diseases.
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Such exposure can lead to transient expression of gene products in blood or formation of granulomas where cells are in contact with choroidal circulation.
“Such an exposure can lead to a transient expression of gene products in blood or the formation of granulomas where cells are in contact with choroidal circulation (35, 36).”
From Subretinal therapy: technological solutions to surgical and immunological challenges (2022) · cites this paper as [35]
The two-step procedure increases the risk of retinotomy enlargement leading to more reflux, and for this reason a one-step approach has been favored in many clinical trials.
“Two-step procedure increases the risk of having the retinotomy enlarge leading to more reflux, for this reason, a one-step approach has been favored in many clinical trials (34–36).”
From Subretinal therapy: technological solutions to surgical and immunological challenges (2022) · cites this paper as [35]
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- Authors
- Weber, M., Rabinowitz, J., Provost, N., Conrath, H., Folliot, S., Briot, D., Chérel, Y., Chenuaud, P., Samulski, J., Moullier, P., & Rolling, F.
- Affiliations
- Inserm (FR)
- University of North Carolina at Chapel Hill (US)
- Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement (FR)
- Corresponding author
- Rolling, Fabienne
- ORCID
- Weber, Michel
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2003-06
- Volume
- 7
- Issue
- 6
- Pages
- 774-781
- ISSN
- 1525-0016 (ISSN-L)
- OpenAlex
- W2162857177
- Cited by
- 203
- References
- 47
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 6 · 2023: 6 · 2022: 4 · 2021: 5 · 2020: 9 · 2019: 5 · 2018: 3 · 2017: 8 · 2016: 1 · 2015: 12 · 2014: 12 · 2013: 9 · 2012: 13
- FWCI
- 7.58
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
- HAL (submittedVersion)
- UNC Libraries
- Funders
- Lions Clubs International Foundation
- Institut National de la Santé et de la Recherche Médicale
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- recombinant adeno-associated virus, retinal pigment epithelium, subretinal gene delivery, retinal transduction, photoreceptors, non-human primates, dogs, rats, Leber congenital amaurosis, retinitis pigmentosa, MERTK, gene therapy vectors
- MeSH terms
- Dependovirus, Animals, Cells, Cultured, Cytomegalovirus, Defective Viruses, DNA, Recombinant, Dogs, Gene Expression Regulation, Genetic Vectors, Luminescent Proteins, Macaca fascicularis, Pigment Epithelium of Eye, Transduction, Genetic, Polymerase Chain Reaction, Rats, Wistar, Transgenes, Photoreceptor Cells, Vertebrate, Green Fluorescent Proteins, Rats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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