Circuit-specific, chemogenetic neuromodulation in nonhuman primates.

摘要摘要UG3/UH3 脑深部电刺激（DBS），适用于丘脑底核（ESTA）等区域，是一种标准的治疗方法， 然而，对于帕金森病（PD），DBS具有固有的手术风险以及感染和 不良副作用我们的首要目标是建立新的化学遗传神经调节策略， 非人灵长类动物（NHP）利用并建立在DBS的优势之上，但解决了许多DBS的局限性， 并最终将其转化为人类的临床治疗。我们专注于设计师专用插座 由设计药物（DREADDs）激活，通过专门的兴奋性或抑制性受体发挥作用 基因植入神经元我们的研究计划，其中集成了一个强大的计划，以提高多样化 Perspectives有一个工具开发（UG3）阶段，然后是临床前试验（UH3）阶段。主要 UG3的目标是：a）在NHP中开发更有效和特异性的DREADD诱导：a）使用 回路特异性逆转录感染方法，选择性感染神经元，包括β-淀粉样蛋白→ GP途径，认为 是PD中运动症状的关键，和B）在手术递送病毒构建体之前使用聚焦超声 以增加DREAD表达在β → GP回路中。我们将使用正电子发射断层扫描（PET）， 行为评估，以衡量可行的DREADD受体表达的强度， 组织学检查以筛选神经病理学并评估转导的神经细胞的密度和解剖分布。 神经元是否决定转移到UH3将基于：1）电机的证据 DREADD激活导致的异常或行为变化，效应量≥ 0.80，和/或2）组织学 DREADD表达的证据，在≥ 35%的神经元中。UH3的主要目标是 使用口服DREADD激动剂去氯氯氮平， (DCZ)减少用神经毒素处理以诱导PD样病症的NHP的运动异常。我们将开始 (AIM 1）通过确定MPTP NHP中DREADD激活的最佳口服激动剂剂量和功效。 然后，我们将（AIM 2）确定口服DREADD激活MPTP NHP的长期疗效和安全性， 临床/行为分析强调临床益处和运动/非运动副作用。我们将使用 在相同的NHP中进行PET，以监测DREADD对微流控电路的影响的稳定性， 组织学分析，以确认DREADD分布在不同回路中，寻找潜在的组织损伤 并确认神经毒素治疗导致的纹状体多巴胺耗竭。最后（目标3），我们将探讨 在相同的NHP中使用PET作为DREADD激动剂单独或作为 联合治疗的一部分，以指导未来人体研究中的剂量调整。为了增强严谨性， 重现性，关键UH3实验将独立验证。这项工作的成功及其人类 翻译可能会改变PD和其他神经/精神疾病的治疗。

ABSTRACT ‒ UG3/UH3 Deep Brain Stimulation (DBS), applied to areas like the subthalamic nucleus (STN), is a standard treatment for Parkinson Disease (PD), however, DBS has inherent surgical risks as well as potential for infections and adverse side effects. Our overarching goal is to establish novel chemogenetic neuromodulation strategies in nonhuman primates (NHPs) that utilize and build upon the strengths of DBS but resolve many DBS limitations, and ultimately to translate these to clinical therapies in humans. We focus on Designer Receptors Exclusively Activated by Designer Drugs (DREADDs), which work via specialized excitatory or inhibitory receptors genetically inserted into neurons. Our Research Plan, which integrates a robust Plan to Enhance Diverse Perspectives, has a tool development (UG3) phase followed by a pre-clinical trial (UH3) phase. The main objective of the UG3 is to: a) develop more effective and specific DREADD induction in NHPs: a) using a circuit-specific retro-infection method to selectively infect the neurons comprising STN→GP pathway, believed to be key to motor symptoms in PD, and b) use focused ultrasound prior to surgical delivery of viral constructs to augment DREAD expression in the STN→GP circuit. We will use positron emission tomography (PET) and behavioral assessments to gauge the strength of viable DREADD receptor expression, and post-mortem histology to screen for neuropathology and to assess the density and anatomical distribution of transduced neurons. The go/no-go for decision for moving to the UH3 will be based on: 1) Evidence of motor abnormalities or behavioral change due to DREADD activation with an effect size ≥0.80, and/or 2) histological evidence of DREADD expression in ≥35% of neurons in the STN. The main objective of the UH3 is to determine if activation of DREADDs in STN neurons, using the oral DREADD agonist deschloroclozapine (DCZ), reduces motor abnormalities NHPs treated with a neurotoxin to induce a PD-like condition. We will start (AIM 1) by determining the optimal oral agonist dosage and efficacy for DREADD activation in MPTP NHPs. We will then (AIM 2) determine the long-term efficacy and safety of oral DREADD activation in MPTP NHPs, with clinical/behavioral analyses emphasizing clinical benefits, and motor/non-motor side effects. We will use PET in the same NHPs to monitor the stability of DREADD effects on STN circuits and follow up with histological analysis to confirm DREADD distribution across STN circuits, to look for potential tissue damage and to confirm striatal dopamine depletion due to the neurotoxin treatment. Finally (AIM 3) we will explore the use of PET in the same NHPs as a noninvasive a gauge for the efficacy of the DREADD agonist alone or as part of a combined therapy to guide dosage adjustment in future human studies. To enhance the rigor and reproducibility, key UH3 experiments will be independently validated. Success in this work and its human translation may be game-changing for the treatment of PD and other neurological/psychiatric disorders.