Sengupta Abhishek, Chaffiol Antoine, Macé Emilie, Caplette Romain, Desrosiers Mélissa, Lampič Maruša, Forster Valérie, Marre Olivier, Lin John Y, Sahel José-Alain, Picaud Serge, Dalkara Deniz, Duebel Jens
INSERM U968, Paris, France.
Sorbonne Universités UPMC Univ Paris 06 UMR_S 968 Institut de la Vision, Paris, France.
EMBO Mol Med. 2016 Nov 2;8(11):1248-1264. doi: 10.15252/emmm.201505699. Print 2016 Nov.
Targeting the photosensitive ion channel channelrhodopsin-2 (ChR2) to the retinal circuitry downstream of photoreceptors holds promise in treating vision loss caused by retinal degeneration. However, the high intensity of blue light necessary to activate channelrhodopsin-2 exceeds the safety threshold of retinal illumination because of its strong potential to induce photochemical damage. In contrast, the damage potential of red-shifted light is vastly lower than that of blue light. Here, we show that a red-shifted channelrhodopsin (ReaChR), delivered by AAV injections in blind rd1 mice, enables restoration of light responses at the retinal, cortical, and behavioral levels, using orange light at intensities below the safety threshold for the human retina. We further show that postmortem macaque retinae infected with AAV-ReaChR can respond with spike trains to orange light at safe intensities. Finally, to directly address the question of translatability to human subjects, we demonstrate for the first time, AAV- and lentivirus-mediated optogenetic spike responses in ganglion cells of the postmortem human retina.
将光敏离子通道——通道视紫红质-2（ChR2）靶向光感受器下游的视网膜回路，有望治疗由视网膜变性引起的视力丧失。然而，激活通道视紫红质-2所需的高强度蓝光超过了视网膜照明的安全阈值，因为它具有很强的诱导光化学损伤的潜力。相比之下，红移光的损伤潜力远低于蓝光。在这里，我们表明，通过向失明的rd1小鼠注射腺相关病毒（AAV）递送的红移通道视紫红质（ReaChR），使用低于人类视网膜安全阈值强度的橙光，能够在视网膜、皮层和行为水平上恢复光反应。我们进一步表明，感染了AAV-ReaChR的猕猴死后视网膜能够以安全强度的橙光激发的脉冲序列做出反应。最后，为了直接解决向人类受试者转化的问题，我们首次证明了在死后人类视网膜神经节细胞中，AAV和慢病毒介导的光遗传学脉冲反应。
