Circuit-specific, chemogenetic neuromodulation in nonhuman primates.

摘要-UG3/UH3 深部脑刺激(DBS)应用于丘脑底核(STN)等区域，是一种标准的治疗方法 然而，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发生神经调节策略 非人灵长类(HAP)利用和建立DBS的优势，但解决了DBS的许多限制， 并最终将这些转化为人类的临床疗法。我们只专注于设计师的受体 由特制药物(DREADD)激活，这种药物通过特殊的兴奋性或抑制性受体发挥作用 通过基因植入神经元。我们的研究计划，其中整合了一个强大的计划，以提高多样化 展望，有一个工具开发(UG3)阶段，然后是临床前试验(UH3)阶段。主 UG3的目标是：a)在NHP中发展更有效和更特异的DREADD诱导：a)使用 电路特异性逆转录感染方法选择性感染含→-gp通路的神经元 是帕金森病患者运动症状的关键，以及b)在手术输送病毒载体之前使用聚焦超声 目的：增强STN→GP回路中DREAD的表达。我们将使用正电子发射断层扫描(PET)和 行为评估，以衡量活的DREADD受体表达的强度，以及尸检 组织学筛查神经病理并评估转导细胞的密度和解剖分布 神经元。决定是否搬到UH3的依据是：1)机动证据 DREADD激活引起的异常或行为改变，其影响大小为≥0.80，和/或2)组织学 ≥中35%的神经元有DREADD的表达。UH3的主要目标是 用口服DREADD激动剂地氯氮平确定STN神经元DREADD的激活情况 (DCZ)，减少用神经毒素治疗的NHP的运动异常，以诱导帕金森病样情况。我们要开始了 (目的1)确定MPTP NHP中DREADD激活的最佳口服激动剂剂量和疗效。 然后我们将(目标2)确定口服DREADD在MPTP NHP中的长期有效性和安全性， 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中监测DREADD对STN电路的稳定性影响并进行随访 组织学分析以确认DREADD在STN回路中的分布，以寻找潜在的组织损伤 并确认神经毒素治疗导致纹状体多巴胺耗竭。最后(目标3)我们将探索 在相同的NHP中使用PET作为无创的DREADD激动剂单独或作为疗效的衡量标准 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。为了加强严谨和 重复性，关键的UH3实验将被独立验证。在这项工作和它的人的成功 翻译可能会改变帕金森氏症和其他神经/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.