Resolving differences between clinical opioids at single neurons

解决临床阿片类药物在单个神经元上的差异

• 批准号: 10355433
• 负责人: Elyssa Margolis
• 金额: $ 20.19万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10355433
• 关键词: Absence of pain sensation; Acute; Affect; Agonist; Alcohol consumption; Analgesics; Animals; Behavioral; Biological; Biological Assay; Biology; Brain; Brain region; Buprenorphine; Cells; Clinical; Complex; Constipation; Data Collection; Detection; Dose; Drug Kinetics; Electrophysiology (science); Euphoria; Evaluation; Fentanyl; Fire - disasters; Future; Habenula; Human; Individual; Investigation; Ligand Binding; Ligands; Measures; Mediating; Morphine; Morphine Dependence; Neurons; Opioid; Opioid Analgesics; Opioid agonist; Oxycodone; Pain; Peptides; Pharmaceutical Preparations; Pharmacology; Placebos; Plague; Property; Protein Kinase; Rattus; Receptor Activation; Recording of previous events; Reporting; Respiration; Rewards; Sampling; Signal Pathway; Signal Transduction; Site; Slice; Standardization; Structure; System; Testing; Variant; Ventilatory Depression; Ventral Tegmental Area; Whole-Cell Recordings; Work; alcohol use disorder; austin; behavioral outcome; clinical development; delta opioid receptor; experience; experimental study; extracellular; high throughput screening; improved; in vivo; individual response; interest; mu opioid receptors; multi-electrode arrays; neural circuit; novel strategies; opioid abuse; opioid overdose; opioid use; opioid use disorder; overexpression; pain relief; preclinical study; receptor; receptor function; response; side effect; therapeutically effective

PROJECT SUMMARY Agonists of the mu opioid receptor (MOR) are currently the most effective pain-relieving drugs, but opioid abuse and overdose continues to plague the USA. The long history of clinical opioid use provides evidence that these drugs differ in underlying biology. For instance, a lack of complete cross tolerance between analgesics and that specific side effects are more intense in response to one drug compared to another (Smith and Peppin, 2014) suggests that they do not function identically at the level of neural circuits. Further, recent animal studies demonstrate a lack of cross tolerance between morphine and fentanyl when these drugs are microinjected directly into the pain modulation circuit (Bobeck et al., 2012, 2019). Ligand bias is the current, dominant hypothesis for how this might happen, however this idea is at best incomplete (Austin Zamarripa et al., 2018; Conibear and Kelly, 2019; Yudin and Rohacs, 2019). The overarching objective of this proposal is to lay a new groundwork for understanding these compounds using responses in neurons from the circuits that contribute to the different in vivo effects of opioids. I have previously demonstrated with whole cell recordings from brain slices, neurons in the ventral tegmental area (VTA) show independent responding to delta opioid receptor agonists that lack in vivo cross tolerance and differentially affect alcohol consumption (Jiang et al., 1991; Mitchell et al., 2014; Margolis et al., 2017). This study provides proof of concept observations that electrophysiology can be used to detect differences in pharmacologies. To enable profiling and comparison of a larger number of molecules, here I propose to use multielectrode extracellular recording from acute brain slices to greatly increase the number of neurons we can record from, thereby increasing the statistical power of the approach. We will investigate the neuronal responses to 4 clinical compounds (morphine, fentanyl, oxycodone, and buprenorphine) and use DAMGO, a highly selective MOR agonist used widely in preclinical studies, as an additional comparator. The majority of neurons in brain regions in the reward circuit (VTA), pain-aversion circuit (habenula), and respiration circuit (Pre-Bötzinger Complex) fire spontaneously in acute brain slices, and these brain regions also highly express the MOR. Therefore, we can use this higher throughput approach to measure potency and efficacy of different MOR agonists in each of these brain regions, as well as compare the responses to sequential application of each ligand in individual neurons in each brain region. Further, we will compare results between naïve animals and those made morphine dependent. By improving our understanding of opioid responses of individual neurons in the specific circuits that underly in vivo opioid effects, this work will (1) demonstrate the viability of using this approach to characterize compounds quickly and (2) identify biological variation of MOR function that will enable more effective therapeutic development for clinical problems that involve opioid signaling, including opioid use disorder, alcohol use disorder, and tolerance to pain medications.

项目摘要 μ阿片受体（莫尔）激动剂是目前最有效的镇痛药物，但阿片受体激动剂是目前最有效的镇痛药物。 滥用和过量继续困扰着美国。阿片类药物临床使用的悠久历史提供了证据 这些药物在基础生物学上是不同的。例如，缺乏完全交叉耐受性， 镇痛药，并且与另一种药物相比，对一种药物的反应更强烈（Smith 和Peppin，2014）表明它们在神经回路水平上的功能并不相同。此外，最近 动物研究表明，当这些药物被 直接微注射到疼痛调制回路中（Bobeck等人，2012年、2019年）。配体偏置是电流， 关于这可能如何发生的主要假设，然而这种想法充其量是不完整的（Austin Zamarripa et 例如，2018; Conibear和Kelly，2019; Yudin和Rohacs，2019）。本提案的总体目标是 为利用神经元的反应来理解这些化合物奠定了新的基础， 有助于阿片类药物的不同体内作用。我之前已经用整个细胞的记录证明了 在脑片上，腹侧被盖区（VTA）的神经元对δ阿片样物质表现出独立的反应 缺乏体内交叉耐受性并差异性地影响酒精消耗的受体激动剂（Jiang等， 1991; Mitchell等人，2014; Margolis等人，2017年）。这项研究提供了概念观察的证据， 电生理学可用于检测药理学的差异。启用分析和比较 更多的分子，在这里我建议使用多电极细胞外记录急性脑 切片，以大大增加我们可以记录的神经元数量，从而增加统计能力， 方法。我们将研究神经元对4种临床化合物（吗啡，芬太尼， 羟考酮和丁丙诺啡），并使用DAMGO，一种在临床前广泛使用的高选择性莫尔激动剂。 研究，作为一个额外的比较。大脑中奖赏回路（VTA）区域的大多数神经元， 疼痛厌恶回路（缰）和呼吸回路（前Bötzinger复合体）在急性发作时自发放电。 大脑切片，这些大脑区域也高度表达莫尔。因此，我们可以使用更高的 通量方法来测量不同莫尔激动剂在这些脑中的每一个中的效力和功效 区域，以及比较每个配体在单个神经元中顺序应用的反应， 每个脑区此外，我们将比较实验动物和吗啡动物的结果 依赖。通过提高我们对特定回路中单个神经元阿片样物质反应的理解， 这是体内阿片类药物作用的基础，这项工作将（1）证明使用这种方法来 快速表征化合物和（2）识别莫尔功能的生物学变异， 针对涉及阿片类信号传导的临床问题的有效治疗开发，包括阿片类药物的使用 障碍，酒精使用障碍，以及对止痛药的耐受性。