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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.

The purpose of our study was to determine changes in amplitudes and implicit times of retinal and cortical pattern evoked potentials with increasing body weight in young, growing rhesus macaques (Macaca mulatta). Retinal and cortical pattern evoked potentials were recorded from 29 male rhesus macaques between 3 and 7 years of age. Thirteen animals were reexamined after 11 months. Computed tomography (CT) was performed on two animals to measure the distance between the location of the skin electrode and the surface of the striate cortex. Spearman correlation coefficients were calculated to describe the relationship between body weights and either root mean square (rms) amplitudes or implicit times. For 13 animals rms amplitudes and implicit times were compared with the Wilcoxon matched pairs signed rank test for recordings taken 11 months apart. Highly significant correlations between increases in body weights and decreases in cortical rms amplitudes were noted in 29 monkeys (p < 0.0005). No significant changes were found in the cortical rms amplitudes in thirteen monkeys over 11 months. Computed tomography showed a large increase of soft tissue thickness over the skull and striate cortex with increased body weight. The decreased amplitude in cortical evoked potentials with weight gain associated with aging can be explained by the increased distance between skin electrode and striate cortex due to soft tissue thickening (passive attenuation).