Safety in nonhuman primates of ocular AAV2-RPE65, a candidate treatment for blindness in leber congenital amaurosis
Safety in nonhuman primates of ocular AAV2-RPE65, a candidate treatment for blindness in leber congenital amaurosis
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DOI：
10.1089/hum.2006.17.845
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发表时间：
2006-08-01
影响因子：
4.2
通讯作者：
Hauswirth, William W.
中科院分区：
文献类型：
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作者：
Jacobson, Samuel G.;Boye, Sanford L.;Hauswirth, William W.
莱伯先天性黑蒙（LCA）是一种导致失明的分子异质性疾病。 RPE65 突变引起的 LCA 已在动物模型中进行了研究，并且通过视网膜下递送 AAV-RPE65 载体恢复了视力。人类眼部基因转移试验正在考虑中。我们对 RPE65 突变狗视网膜下 AAV-2/2.RPE65 进行的安全性研究表明，在较高载体剂量下，一些动物的注射区域中的光感受器适度丧失。我们现在检验这样一个假设：正常猴子的视网膜与人类相似，可能存在与载体相关的毒性。在正常食蟹猴中单次眼内注射 AAV-2/2.RPE65 后，进行了为期 1 周和 3 个月的良好实验室规范安全性研究。未发现全身毒性。眼部特异性研究包括临床检查、视网膜电图检查和视网膜组织病理学。注射后眼部炎症症状在 1 周时几乎消失。 3 个月时，注射载体的眼睛的视网膜电图与注射载体的对照眼或与术前记录相比没有不同。通过临床和组织病理学记录视网膜下注射的视网膜穿孔愈合部位。视网膜下注射的眼睛、载体或车辆的中心凹结构可能是异常的。中央视网膜的形态测量显示没有以剂量依赖性方式发生感光层厚度异常。 1周时，载体序列存在于注射的视网膜、玻璃体和视神经中，但在大脑中并不一致。 3个月时，视神经和大脑中没有载体序列。结果允许考虑在未来视网膜下 AAV-2/2.RPE65 人体试验中未观察到不良反应水平的上限。中心凹治疗 LCA 和其他视网膜变性的潜在价值值得进一步研究如何实现基因转移而不因视网膜手术脱离造成视网膜损伤。
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 vector-related 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.