title: Long-term doxycycline-regulated transgene expression in the retina of nonhuman primates following subretinal injection of recombinant AAV vectors.
abstract: Adeno-associated viral gene therapy has shown promise for the treatment of inherited and acquired retinal disorders. In most applications, regulation of expression is a critical concern for both safety and efficacy. The purpose of our study was to evaluate the ability of the tetracycline-regulatable system to establish long-term transgene regulation in the retina of nonhuman primates. Three rAAV vectors expressing the tetracycline-dependent transactivator (rtTA) under the control of either the ubiquitous CAG promoter or the specific RPE65 promoter (AAV2/5.CAG.TetOn.epo, AAV2/4.CAG.TetOn.epo, and AAV2/4.RPE65.TetOn.epo) were generated and administered subretinally to seven macaques. We demonstrated that repeated inductions of transgene expression in the nonhuman primate retina can be achieved using a Tet-inducible system via rAAV vector administration over a long period (2.5 years). Maximum erythropoietin (EPO) secretion in the anterior chamber depends upon the rAAV serotype and the nature of the promoter driving rtTA expression. We observed that the EPO isoforms produced in the retina differ from one another based on the transduced cell type of origin within the retina and also differ from both the physiological EPO isoforms and the isoforms produced by AAV-transduced skeletal muscle.
authors.affiliation: INSERM UMR U649, CHU-Hotel Dieu, Bât. J. Monnet, 30 Avenue J. Monnet, 44035 Nantes Cedex 01, France.
authors.forename: Knut
authors.initials: K
authors.lastname: Stieger
authors.forename: Guylène
authors.initials: G
authors.lastname: Le Meur
authors.forename: Françoise
authors.initials: F
authors.lastname: Lasne
authors.forename: Michel
authors.initials: M
authors.lastname: Weber
authors.forename: Jack-Yves
authors.initials: JY
authors.lastname: Deschamps
authors.forename: Delphine
authors.initials: D
authors.lastname: Nivard
authors.forename: Alexandra
authors.initials: A
authors.lastname: Mendes-Madeira
authors.forename: Nathalie
authors.initials: N
authors.lastname: Provost
authors.forename: Laurent
authors.initials: L
authors.lastname: Martin
authors.forename: Philippe
authors.initials: P
authors.lastname: Moullier
authors.forename: Fabienne
authors.initials: F
authors.lastname: Rolling
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology*; D000229:Dependovirus / Q000187:drug effects / Q000235:genetics*; D004318:Doxycycline / Q000378:metabolism / Q000494:pharmacology*; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage*; D008251:Macaca; D012160:Retina / Q000187:drug effects* / Q000378:metabolism; D019076:Transgenes
chemical_list: D000900:Anti-Bacterial Agents; D004318:Doxycycline