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

摘要 光遗传学是神经科学领域的一项革命性技术。通过将光敏蛋白与颅内 通过光传递，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。技术 已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，光遗传学迄今为止未能对使用更大动物的研究产生重大影响 与人类相似，例如猕猴，破坏了其对人类患者的转化潜力。我们 开展了一项世界范围的开放科学计划，以确定灵长类动物尚待解决的挑战 光遗传学（Tremblay 等人 Neuron，2020）。我们确定了猕猴大脑的绝对大小 比小鼠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 表达和光传递。我们的多学科研究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）使用大规模、安全的超敏感视蛋白递送 基因治疗技术； 2）可长期植入、超薄、灵活、生物相容性LED阵列；和 3) 植入式电池供电 LED 驱动器，用于在不受约束的自然行为期间进行无线控制。这个 该方法将允许以细胞类型特异性和毫秒精确控制灵长类动物的大量大脑 在不受身体约束的猴子中解决问题，从而允许对介导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。该技术平台可以 可直接用作患有神经系统疾病的人类的细胞类型特异性光遗传学疗法 影响特定神经群体，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.