Precision pharmacology by local control of opioid receptors in neural circuits

通过局部控制神经回路中的阿片受体实现精准药理学

• 批准号: 10044383
• 负责人: Aashish Manglik
• 金额: $ 44.33万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10044383
• 关键词: Absence of pain sensation; Acute; Agonist; Analgesics; Behavioral; Brain; Breathing; Cytoplasm; Dangerousness; Development; Electrophysiology (science); Enkephalins; G-Protein-Coupled Receptors; Goals; Heroin; Heterogeneity; Hyperalgesia; Location; Mediating; Membrane; Methods; Modeling; Molecular; Morphine; Mus; Neurons; Neuropeptides; Nociception; Opiate Addiction; Opioid; Opioid Peptide; Opioid Receptor; Pathologic; Peptides; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Phase; Physiological; Preparation; Proteins; Psychological reinforcement; Receptor Activation; Receptor Signaling; Recombinant adeno-associated virus (rAAV); Resolution; Series; Signal Transduction; Slice; Spinal Cord; System; Technology; Testing; Therapeutic; Therapeutic Index; Time; Tissues; Ventilatory Depression; base; behavioral response; clinically relevant; comparative efficacy; drug action; efficacy testing; endogenous opioids; high risk; improved; in vivo; insight; instrument; mu opioid receptors; nanobodies; neural circuit; neuroregulation; novel; optogenetics; programs; receptor; receptor function; recruit; side effect; tool

PROJECT SUMMARY This proposal seeks to leverage the emerging understanding of cellular and subcellular heterogeneity in opioid signaling for therapeutic discovery. The project develops new optogenetic and chemogenetic tools to achieve local control of opioid receptor activity and signaling in neural circuits and at a subcellular level of resolution. The proposal seeks to develop a versatile toolbox and provide proof-of-concept for "in vivo precision pharmacology" that have not yet been applied to the opioid system. The first phase (R61) is a "high-risk" technology enabling phase, based on combining our collaborative team's expertise in developing receptor-blocking intracellular nanobodies with established methods for conferring local control of nanobody concentration in the cytoplasm and specific membrane domains, using optogenetic and chemogenetic strategies that have already been validated but never before applied to opioid receptors. The R61 phase will develop these tools (Specific Aim 1) and validate them in neuronal culture and acute slice preparations (Specific Aim 2). These studies will provide not only proof-of-concept for in vivo precision pharmacology of opioid receptors, but also for G protein-coupled receptors as a class. We have set a specific series of milestones to evaluate progress, and to assess feasibility for advance to in vivo studies. If the milestones are successfully met, the second phase (R33) will apply the new tools to selectively target opioid signaling in neural circuits in vivo, focusing on analgesia, behavioral reinforcement and respiratory depression by clinically relevant opioid drugs. Mice are used as a well-established and relevant model (Specific Aim 3). These studies leverage the combined expertise of our collaborative team, and will establish powerful optogenetic and chemogenetic manipulations for the study of in vivo opioid function for the first time. Accordingly, the proposed studies to provide the first-in-class application of optogenetic and chemogenetic control to the opioid system in vivo, and they will explore a precision pharmacology of opioids based on location in neural circuits.

项目概要 该提案旨在利用对阿片类药物细胞和亚细胞异质性的新兴理解 治疗发现的信号传导。该项目开发了新的光遗传学和化学遗传学工具来实现 神经回路中阿片受体活性和信号传导以及亚细胞分辨率的局部控制。这 该提案旨在开发一个多功能工具箱，并为“体内精准药理学”提供概念验证 尚未应用于阿片类药物系统。第一阶段（R61）是一项“高风险”技术 阶段，基于结合我们合作团队在开发受体阻断细胞内药物方面的专业知识 具有对细胞质中纳米抗体浓度进行局部控制的既定方法的纳米抗体 和特定的膜域，使用已经被采用的光遗传学和化学遗传学策略 已得到验证，但之前从未应用于阿片受体。 R61阶段将开发这些工具（具体目标1） 并在神经元培养和急性切片制备中验证它们（具体目标 2）。这些研究将提供 不仅是阿片受体体内精确药理学的概念验证，还包括 G 蛋白偶联的概念验证 受体作为一类。我们设定了一系列具体的里程碑来评估进展并评估可行性 用于推进体内研究。如果成功达到里程碑，第二阶段（R33）将应用新的 选择性靶向体内神经回路中阿片类信号传导的工具，重点关注镇痛、行为 临床相关阿片类药物的强化和呼吸抑制。小鼠被用作成熟的 和相关模型（具体目标 3）。这些研究利用了我们协作团队的综合专业知识， 并将建立强大的光遗传学和化学遗传学操作来研究体内阿片类药物功能 首次。因此，拟议的研究提供了光遗传学和光遗传学的一流应用 对体内阿片类药物系统的化学遗传学控制，他们将探索阿片类药物的精确药理学 基于神经回路中的位置。