Gene therapy restores vision-dependent behavior as well as retinal structure and function in a mouse model of RPE65 Leber congenital amaurosis
Gene therapy restores vision-dependent behavior as well as retinal structure and function in a mouse model of RPE65 Leber congenital amaurosis
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DOI：
10.1016/j.ymthe.2005.09.001
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发表时间：
2006-03-01
影响因子：
12.4
通讯作者：
Hauswirth, WW
中科院分区：
文献类型：
--
作者：
Pang, JJ;Chang, B;Hauswirth, WW
视网膜色素上皮特异性蛋白65 kDa （RPE65）是一种负责全反式视黄醛异构化为光活性11-顺式视黄醛的蛋白质，对视觉循环至关重要。RPE65突变可导致严重的早发性视网膜疾病，如莱伯先天性黑朦（LCA）。自然发生的LCA啮齿动物模型Rpe65隐性无义突变，rd12小鼠，显示杆视网膜电图（ERG）严重减少，缺乏11-顺式视黄醛和视紫红质，视网膜色素上皮（RPE）细胞中视黄酯过度积累，以及光受体变性。rd12小鼠在出生后第14天视网膜下注射rAAV5-CBA-hRPE65载体。治疗后不久，RPE65在大面积RPE上表达。这导致视紫红质水平提高，ERG信号恢复到接近正常水平。视黄醇酯水平维持在接近正常水平，眼底和视网膜形态保持正常。恢复视网膜健康的所有参数至少在7个月内保持稳定。将Morris水迷宫行为实验改进为在极弱光线下测试棒的功能；在一只眼睛上治疗的rd12小鼠表现与正常视力的C57BL/6小鼠相似，而未经治疗的rd12小鼠表现非常差，这表明基因治疗可以恢复先天性失明动物正常的视觉依赖行为。
Retinal pigment epithelium-specific protein 65 kDa (RPE65) is a protein responsible for isomerization of all-trans-retinaldehyde to its photoactive 11-cis-retinaidehyde and is essential for the visual cycle. RPE65 mutations can cause severe, early onset retinal diseases such as Leber congenital amaurosis (LCA). A naturally occurring rodent model of LCA with a recessive nonsense Rpe65 mutation, the rd12 mouse, displays a profoundly diminished rod electroretinogram (ERG), an absence of 11-cis-retinalclehyde and rhodopsin, an overaccumulation of retinyl esters in retinal pigmented epithelial (RPE) cells, and photoreceptor degeneration. rd12 mice were injected subretinally at postnatal day 14 with rAAV5-CBA-hRPE65 vector. RPE65 expression was found over large areas of RPE soon after treatment. This led to improved rhodopsin levels with ERG signals restored to near normal. Retinyl ester levels were maintained at near normal, and fundus and retinal morphology remained normal. All parameters of restored retinal health remained stable for at least 7 months. The Morris water maze behavioral test was modified to test rod function under very dim light; rd12 mice treated in one eye performed similar to normally sighted C57BL/6) mice, while untreated rd12 mice performed very poorly, demonstrating that gene therapy can restore normal vision-dependent behavior in a congenitally blind animal.