Long-Term Preservation of Cones and Improvement in Visual Function Following Gene Therapy in a Mouse Model of Leber Congenital Amaurosis Caused by Guanylate Cyclase-1 Deficiency
Long-Term Preservation of Cones and Improvement in Visual Function Following Gene Therapy in a Mouse Model of Leber Congenital Amaurosis Caused by Guanylate Cyclase-1 Deficiency
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DOI：
10.1089/hum.2011.069
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发表时间：
2011-10-01
影响因子：
4.2
通讯作者：
Ali, Robin R.
中科院分区：
文献类型：
--
作者：
Mihelec, Marija;Pearson, Rachael A.;Ali, Robin R.
Leber先天性黑色素沉着症是一种严重的视网膜营养不良症，从婴儿期开始就表现为视力低下或失明。GUCY2D功能缺失突变导致LCA1，是LCA最常见的原因之一，占所有病例的20%。人GUCY2D和小鼠Gucy2e基因编码鸟苷环化酶-1(GC)，负责在光暴露后恢复光感受器的暗状态。Glicy2e(-/-)小鼠的视杆功能部分减弱，但在变性前没有视锥功能。尽管锥体在形态上看起来是正常的，但它们显示了参与光转导的蛋白质的错误定位。在这项研究中，我们测试了含有人视紫红质激酶启动子和人GUCY2D基因的rAAV2/8载体的效果。在Glicy2e(-/-)小鼠的视网膜下，Gc1蛋白在视杆细胞和视锥细胞的外节段中被检测到，并且在视锥细胞转导蛋白的转导区域中被适当地定位到视锥细胞外节。此外，我们观察到剂量依赖的视杆和视锥功能的恢复以及治疗后小鼠视觉行为的改善。最重要的是，在注射后6个月观察到转移区的锥体保存。到目前为止，这是拯救LCA Glicy2e(-/-)小鼠模型的最有效的方法，我们建议一种类似于本研究中使用的载体，可能适合用于LCA1基因治疗的临床试验。
Leber congenital amaurosis (LCA) is a severe retinal dystrophy manifesting from early infancy as poor vision or blindness. Loss-of-function mutations in GUCY2D cause LCA1 and are one of the most common causes of LCA, accounting for 20% of all cases. Human GUCY2D and mouse Gucy2e genes encode guanylate cyclase-1 (GC), which is responsible for restoring the dark state in photoreceptors after light exposure. The Glicy2e(-/-) mouse shows partially diminished rod function, but an absence of cone function before degeneration. Although the cones appear morphologically normal, they exhibit mislocalization of proteins involved in phototransduction. In this study we tested the efficacy of an rAAV2/8 vector containing the human rhodopsin kinase promoter and the human GUCY2D gene. Following subretinal delivery of the vector in Glicy2e(-/-) mice, GC1 protein was detected in the rod and cone outer segments, and in transduced areas of retina cone transducin was appropriately localized to cone outer segments. Moreover, we observed a dose-dependent restoration of rod and cone function and an improvement in visual behavior of the treated mice. Most importantly, cone preservation was observed in transduced areas up to 6 months post injection. To date, this is the most effective rescue of the Glicy2e(-/-) mouse model of LCA and we propose that a vector, similar to the one used in this study, could be suitable for use in a clinical trial of gene therapy for LCA1.