Safety in nonhuman primates of ocular AAV2-RPE65, a candidate treatment for blindness in Leber congenital amaurosis.
Leber congenital amaurosis (LCA) is a molecularly heterogeneous disease group that leads to blindness. LCA caused by RPE65 mutations has been studied in animal models and vision has been restored by subretinal delivery of AAV-RPE65 vector. Human ocular gene transfer trials are being considered. Our safety studies of subretinal AAV-2/2.RPE65 in RPE65-mutant dogs showed evidence of modest photoreceptor loss in the injection region in some animals at higher vector doses. We now test the hypothesis that there can be vectorrelated toxicity to the normal monkey, with its human-like retina. Good Laboratory Practice safety studies following single intraocular injections of AAV-2/2.RPE65 in normal cynomolgus monkeys were performed for 1-week and 3-month durations. Systemic toxicity was not identified. Ocular-specific studies included clinical examinations, electroretinography, and retinal histopathology. Signs of ocular inflammation postinjection had almost disappeared by 1 week. At 3 months, electroretinography in vector-injected eyes was no different than in vehicle-injected control eyes or compared with presurgical recordings. Healed sites of retinal perforation from subretinal injections were noted clinically and by histopathology. Foveal architecture in subretinally injected eyes, vector or vehicle, could be abnormal. Morphometry of central retina showed no photoreceptor layer thickness abnormalities occurring in a dose-dependent manner. Vector sequences were present in the injected retina, vitreous, and optic nerve at 1 week but not consistently in the brain. At 3 months, there were no vector sequences in optic nerve and brain. The results allow for consideration of an upper range for no observed adverse effect level in future human trials of subretinal AAV-2/2.RPE65. The potential value of foveal treatment for LCA and other retinal degenerations warrants further research into how to achieve gene transfer without retinal injury from surgical detachment of the retina.
Jacobson
Samuel G
SG
Department of Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA 19104, USA. jacobsos@mail.med.upenn.edu
Boye
Sanford L
SL
Aleman
Tomas S
TS
Conlon
Thomas J
TJ
Zeiss
Caroline J
CJ
Roman
Alejandro J
AJ
Cideciyan
Artur V
AV
Schwartz
Sharon B
SB
Komaromy
Andras M
AM
Doobrajh
Michelle
M
Cheung
Andy Y
AY
Sumaroka
Alexander
A
Pearce-Kelling
Susan E
SE
Aguirre
Gustavo D
GD
Kaushal
Shalesh
S
Maguire
Albert M
AM
Flotte
Terence R
TR
Hauswirth
William W
WW
D000818:Animals; D001766:Blindness / Q000209:etiology / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D001921:Brain / Q000378:metabolism / Q000473:pathology / Q000821:virology; D002352:Carrier Proteins; D000229:Dependovirus; D004285:Dogs; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D015316:Genetic Therapy; D006801:Humans; D008252:Macaca fascicularis; D009154:Mutation; D029242:Optic Atrophy, Hereditary, Leber / Q000150:complications / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D009900:Optic Nerve / Q000378:metabolism / Q000473:pathology / Q000821:virology; D010786:Photoreceptor Cells / Q000378:metabolism / Q000473:pathology / Q000821:virology; D012160:Retina / Q000378:metabolism / Q000473:pathology / Q000821:virology; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
D002352:Carrier Proteins; D005136:Eye Proteins; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase