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.
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.
Weber
Michel
M
Laboratoire de Thérapie Génique, INSERM ERM01-05, CHU-Hotel DIEU, Bat. J. Monnet, 30 Avenue J. Monnet, 44035, Nantes Cedex 01, France.
Rabinowitz
Joseph
J
Provost
Nathalie
N
Conrath
Hervé
H
Folliot
Sébastien
S
Briot
Delphine
D
Chérel
Yan
Y
Chenuaud
Pierre
P
Samulski
Jude
J
Moullier
Philippe
P
Rolling
Fabienne
F
D000818:Animals; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics; D004274:DNA, Recombinant; D003673:Defective Viruses; D000229:Dependovirus / Q000145:classification / Q000235:genetics*; D004285:Dogs; D005786:Gene Expression Regulation / Q000502:physiology*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008252:Macaca fascicularis; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism*; D010857:Pigment Epithelium of Eye / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D051381:Rats; D017208:Rats, Wistar; D014161:Transduction, Genetic; D019076:Transgenes
D004274:DNA, Recombinant; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins