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。通过将光敏蛋白与颅内结合 在光传递方面，光遗传学对神经元活动提供了前所未有的、细胞类型的特定控制。这项技术 已经成为研究小动物模型(如老鼠和苍蝇)神经回路的主要方法。 不幸的是，光遗传学到目前为止还没有对更多使用更大动物的研究产生重大影响。 类似于人类，如猕猴，破坏了其对人类患者的翻译潜力。我们 开展了一项世界范围的开放科学倡议，以确定灵长类中有待解决的挑战 光遗传学(Tremblay等人)神经元，2020)。我们确定了猕猴大脑的绝对大小，这是 比小鼠的大脑及其免疫系统大200倍，这是这两个基因面临的主要挑战 表情和轻盈的传递。我们的多学科调查团队将通过以下方式克服这些障碍 开发和优化三项新技术：1)大规模、安全地输送超灵敏光学元件 基因治疗技术；2)长期植入、超薄、灵活、生物兼容的发光二极管阵列；以及3) 可植入电池供电的LED驱动器，可在自由、自然的行为中进行无线控制。这 这种方法将允许精确控制大量灵长类动物的大脑，具有细胞类型特异性和毫秒级 在猴子中不受身体约束的分解，从而允许对调节的神经回路进行因果解剖 与理解和治疗人类大脑疾病相关的自然行为。这一技术平台可以 作为一种针对细胞类型的光遗传疗法，直接应用于患有神经疾病的人类 影响特定神经群体的疾病，例如局灶性癫痫。

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.