title: Safety of recombinant adeno-associated virus type 2-RPE65 vector delivered by ocular subretinal injection.
abstract: AAV2 delivery of the RPE65 gene to the retina of blind RPE65-deficient animals restores vision. This strategy is being considered for human trials in RPE65-associated Leber congenital amaurosis (LCA), but toxicity and dose efficacy have not been defined. We studied ocular delivery of AAV-2/2.RPE65 in RPE65-mutant dogs. There was no systemic toxicity. Ocular examinations showed mild or moderate inflammation that resolved over 3 months. Retinal histopathology indicated that traumatic lesions from the injection were common, but thinning within the injection region occurred only at the two highest vector doses. Biodistribution studies at 3 months postinjection showed no vector in optic nerve or visual centers in the brain and only isolated non-dose-related detection in other organs. We also performed biodistribution studies in normal rats at about 2 weeks and 2 months postinjection and vector was not widespread outside the injected eye. Dose-response results in RPE65-mutant dogs indicated that the highest 1.5-log unit range of vector doses proved efficacious. The efficacy and toxicity limits defined in this study lead to suggestions for the design of a subretinal AAV-2/2.RPE65 human trial of RPE65-associated LCA.
authors.affiliation: Scheie Eye Institute, Department of Ophthalmology, University of Pennsylvania, Philadelphia, PA 19104, USA. jacobsos@mail.med.upenn.edu
authors.forename: Samuel G
authors.initials: SG
authors.lastname: Jacobson
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Acland
authors.forename: Gustavo D
authors.initials: GD
authors.lastname: Aguirre
authors.forename: Tomas S
authors.initials: TS
authors.lastname: Aleman
authors.forename: Sharon B
authors.initials: SB
authors.lastname: Schwartz
authors.forename: Artur V
authors.initials: AV
authors.lastname: Cideciyan
authors.forename: Caroline J
authors.initials: CJ
authors.lastname: Zeiss
authors.forename: Andras M
authors.initials: AM
authors.lastname: Komaromy
authors.forename: Shalesh
authors.initials: S
authors.lastname: Kaushal
authors.forename: Alejandro J
authors.initials: AJ
authors.lastname: Roman
authors.forename: Elizabeth A M
authors.initials: EA
authors.lastname: Windsor
authors.forename: Alexander
authors.initials: A
authors.lastname: Sumaroka
authors.forename: Susan E
authors.initials: SE
authors.lastname: Pearce-Kelling
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Conlon
authors.forename: Vincent A
authors.initials: VA
authors.lastname: Chiodo
authors.forename: Sanford L
authors.initials: SL
authors.lastname: Boye
authors.forename: Terence R
authors.initials: TR
authors.lastname: Flotte
authors.forename: Albert M
authors.initials: AM
authors.lastname: Maguire
authors.forename: Jean
authors.initials: J
authors.lastname: Bennett
authors.forename: William W
authors.initials: WW
authors.lastname: Hauswirth
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001766:Blindness / Q000235:genetics / Q000628:therapy; D002352:Carrier Proteins; D000229:Dependovirus / Q000235:genetics*; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D064420:Drug-Related Side Effects and Adverse Reactions; D005136:Eye Proteins / Q000008:administration & dosage / Q000235:genetics*; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000009:adverse effects* / Q000493:pharmacokinetics; D015552:Injections, Intralesional; D008297:Male; D029242:Optic Atrophy, Hereditary, Leber / Q000235:genetics / Q000628:therapy; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics; D012160:Retina / Q000187:drug effects* / Q000473:pathology; D014018:Tissue Distribution; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
chemical_list: D002352:Carrier Proteins; D005136:Eye Proteins; D011994:Recombinant Proteins; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase