Noninvasive gene delivery to foveal cones for vision restoration
Noninvasive gene delivery to foveal cones for vision restoration
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DOI：
10.1172/jci.insight.96029
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发表时间：
2018-01-25
期刊：
影响因子：
8
通讯作者：
Dalkara, Deniz
中科院分区：
文献类型：
--
作者：
Khabou, Hanen;Garita-Hernandez, Marcela;Dalkara, Deniz
眼内注射腺相关病毒（AAV）载体是将基因药物输送到视网膜的明显途径。然而，我们对灵长类动物眼睛视网膜下和玻璃体内空间的解剖学独特环境中的 AAV 转导模式的理解存在差距，阻碍了在临床上建立无创且有效的基因传递至中心凹锥体的方法。在这里，我们建立了新的载体-启动子组合，以克服与 AAV 介导的中央凹视锥细胞转导相关的局限性，并支持小鼠模型、人类诱导多能干细胞衍生的类器官、死后人类视网膜外植体和活体猕猴的研究。我们证明，当将 AAV9 变体注射到距中央凹几毫米的视网膜下空间时，可以提供有效的中央凹锥体转导，而不会分离这个脆弱的区域。工程化的 AAV2 变体以玻璃体内施用的耐受性良好的剂量将基因递送至中央凹锥体。两种递送方式都依赖于视锥特异性启动子，并导致与光遗传学视力恢复兼容的高水平转基因表达。这里描述的模型系统提供了对跨物种 AAV 载体行为的深入了解，以获得神经退行性疾病基因治疗所需的安全性和有效性。
Intraocular injection of adeno-associated viral (AAV) vectors has been an evident route for delivering gene drugs into the retina. However, gaps in our understanding of AAV transduction patterns within the anatomically unique environments of the subretinal and intravitreal space of the primate eye impeded the establishment of noninvasive and efficient gene delivery to foveal cones in the clinic. Here, we establish new vector-promoter combinations to overcome the limitations associated with AAV-mediated cone transduction in the fovea with supporting studies in mouse models, human induced pluripotent stem cell-derived organoids, postmortem human retinal explants, and living macaques. We show that an AAV9 variant provides efficient foveal cone transduction when injected into the subretinal space several millimeters away from the fovea, without detaching this delicate region. An engineered AAV2 variant provides gene delivery to foveal cones with a well-tolerated dose administered intravitreally. Both delivery modalities rely on a cone-specific promoter and result in high-level transgene expression compatible with optogenetic vision restoration. The model systems described here provide insight into the behavior of AAV vectors across species to obtain safety and efficacy needed for gene therapy in neurodegenerative disorders.