Sensitization of ON-bipolar cells with ambient light activatable multi-characteristic opsin rescues vision in mice.
Sensitization of ON-bipolar cells with ambient light activatable multi-characteristic opsin rescues vision in mice.
复制标题
DOI：
10.1038/s41434-020-00200-2
复制
发表时间：
2021-04
期刊：
影响因子：
5.1
通讯作者：
Mohanty, Samarendra
中科院分区：
文献类型：
--
作者：
Batabyal, Subrata;Gajjeraman, Sivakumar;Pradhan, Sanjay;Bhattacharya, Sulagna;Wright, Weldon;Mohanty, Samarendra
基于基因治疗的治疗，如光遗传学，提供了一种潜在的强大的方法来绕过视网膜退行性疾病中受损的光感受器，并使用剩余的视网膜细胞进行功能化以实现光敏性。然而，目前的光遗传学治疗方法依赖于需要高强度光来激活的视蛋白，因此增加了用作可穿戴设备的一部分的挑战。在这里，我们报告了AAV 2辅助递送高度光敏的多特征视蛋白（MCO 1）到视网膜变性小鼠的ON双极细胞中，以允许通过环境光激活。通过不同剂量的携带MCO 1（vMCO 1）的AAV 2到靶细胞中的递送功效的严格表征显示递送后6个月内的持久表达，如通过报告基因表达所测量的。持久的MCO 1表达与显著改善的行为结果相关，这是纵向测量的视觉水迷宫和视运动试验。vMCO 1注射组血浆和玻璃体液中的促炎/抗炎细胞因子水平与基线或对照组相比无显著变化。此外，在注射不同剂量vMCO 1的动物组中，注射后不同时间点的生物分布研究显示，在非靶组织中未检出载体拷贝。vMCO 1转染视网膜组织的免疫组化显示MCO 1的双极特异性表达和免疫/炎症反应的缺乏。此外，使用SDOCT的眼部成像显示vMCO 1注射眼的结构架构没有变化。在患有视网膜变性的受试者中诱导对剩余的健康双极细胞的环境光响应性将允许视网膜回路获得视敏度而不需要主动刺激装置。
Gene therapy-based treatment such as optogenetics offers a potentially powerful way to bypass damaged photoreceptors in retinal degenerative diseases and use the remaining retinal cells for functionalization to achieve photosensitivity. However, current approaches of optogenetic treatment rely on opsins that require high intensity light for activation thus adding to the challenge for use as part of a wearable device. Here, we report AAV2 assisted delivery of highly photosensitive multi-characteristic opsin (MCO1) into ON bipolar cells of mice with retinal degeneration to allow activation by ambient light. Rigorous characterization of delivery efficacy by different doses of AAV2 carrying MCO1 (vMCO1) into targeted cells showed durable expression over 6 months after delivery as measured by reporter expression. The enduring MCO1 expression was correlated with the significantly improved behavioral outcome, that was longitudinally measured by visual water maze and optomotor assays. The pro/anti-inflammatory cytokine levels in plasma and vitreous humor of the vMCO1 injected group did not change significantly from baseline or control group. Furthermore, biodistribution studies at various time points after injection in animal groups injected with different doses of vMCO1 showed non-detectable vector copies in non-targeted tissues. Immunohistochemistry of vMCO1 transfected retinal tissues showed bipolar specific expression of MCO1 and the absence of immune/inflammatory response. Furthermore, ocular imaging using SDOCT showed no change in the structural architecture of vMCO1-injected eyes. Induction of ambient light-responsiveness to remaining healthy bipolar cells in subjects with retinal degeneration will allow the retinal circuitry to gain visual acuity without requiring an active stimulation device.
登录
查看更多内容
影响因子：
4.4
作者：
de Balthasar, Chloe;Patel, Sweta;Fine, Ione
通讯作者：
Fine, Ione
影响因子：
4.4
作者：
Brankatschk, Robert;Bodenhausen, Natacha;Buergmann, Helmut
通讯作者：
Buergmann, Helmut
影响因子：
1.8
作者：
Humayun, MS;Weiland, JD;de Juan, E
通讯作者：
de Juan, E
影响因子：
1.9
作者：
Hetling, JR;Baig-Silva, MS
通讯作者：
Baig-Silva, MS
影响因子：
4.4
作者：
Liu, Yang;McDowell, Colleen M.;Clark, Abbot F.
通讯作者：
Clark, Abbot F.