Optimizing Optogenetics for Cell-type-specific Control in Freely-moving Primates

摘要 光遗传学是神经科学中的一项革命性技术。通过结合光敏蛋白和颅内 光传递，光遗传学提供了前所未有的，对神经元活动的细胞类型特异性控制。该技术 已经成为研究小鼠和苍蝇等小动物模型神经回路的主要方法。 不幸的是，光遗传学迄今未能对更多使用大型动物的研究产生重大影响。 与人类相似，如猕猴，破坏了其对人类患者的转化潜力。我们 开展了一项全球性的开放科学计划，以确定灵长类动物中有待解决的挑战。 光遗传学（Tremblay等人Neuron，2020）。我们确定了猕猴大脑的绝对大小， 比老鼠的大脑大200倍，免疫系统也是如此，这是两个基因的主要挑战。 表达和光传递。我们的多学科研究团队将克服这些障碍， 开发和优化三项新技术：1）大规模，安全输送超敏感视蛋白， 基因治疗技术; 2）可长期植入的、超薄的、柔性的、生物相容的LED阵列;以及3） 可植入的电池供电LED驱动器，用于在不受限制的自然行为期间进行无线控制。这 这种方法将允许精确控制大体积的灵长类动物的大脑与细胞类型特异性和毫秒 解决猴子自由的身体约束，从而允许因果解剖的神经回路介导 与理解和治疗人类大脑疾病相关的自然行为。这个技术平台可以 可直接作为细胞类型特异性光遗传学疗法应用于患有神经障碍的人 影响特定的神经群体，如局灶性癫痫。

ABSTRACT Optogenetics is a revolutionary technique in neuroscience. By combining light-sensitive proteins with intracranial light delivery, optogenetics offers unprecedented, cell-type specific control over neuronal activity. The technique has become the dominant approach for studying neural circuits in small animal models such as mice and flies. Unfortunately, optogenetics has so far failed to have a major impact on research using larger animals more similar to humans, such as macaque monkeys, undermining its translational potential for human patients. We conducted a world-wide Open Science initiative to identify the challenges remaining to be solved in primate optogenetics (Tremblay et al. Neuron, 2020). We identified the sheer size of the macaque monkey brain, which is 200 times bigger than the mouse brain, as well as its immune system, as the main challenges for both gene expression and light delivery. Our multidisciplinary team of investigators will overcome these obstacles by developing and optimizing three new technologies: 1) large-scale, safe delivery of ultra-sensitive opsins using gene therapy techniques; 2) chronically-implantable, ultra-thin, flexible, biocompatible LED arrays; and 3) implantable, battery-powered LED drivers for wireless control during unrestrained, naturalistic behavior. This approach will allow precise control of large volumes of the primate brain with cell-type specificity and millisecond resolution in monkeys free of physical restraint, thus permitting causal dissection of the neural circuits mediating natural behavior relevant for understanding and treating human brain disorders. This technology platform could be directly applied as a cell-type-specific optogenetic therapy for humans suffering from neurological disorders that affect specific neural populations, such as focal epilepsy.