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

基本信息

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
项目类别：
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

项目摘要

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 MU阿片受体（MOR）的激动剂目前是最有效的疼痛药物，但是阿片类药物滥用和过量的人继续困扰着美国。临床阿片类药物使用的悠久历史提供了证据这些药物在潜在的生物学上有所不同。例如，之间缺乏完全的交叉耐受性与另一种药物相比，镇痛药和特定的副作用更为强烈和Peppin，2014年）表明，它们在神经回路水平上的功能不相同。此外，最近动物研究表明，当这些药物是吗啡和芬太尼之间缺乏交叉耐受性显微注射直接进入疼痛调节电路（Bobeck等，2012，2019）。配体偏见是电流，对于这种情况可能发生的主要假设，但是这个想法充其量是不完整的（Austin Zamarripa et Al。，2018; Conibear and Kelly，2019年； Yudin和Rohacs，2019年）。该提议的总体目标是为使用电路中的神经元中的反应来理解这些化合物的新基础，以了解这些化合物有助于阿片类药物的体内影响不同。我以前已经证明了全细胞记录从脑切片中，腹侧对盖区域（VTA）的神经元显示出独立响应三角洲的响应缺乏体内跨耐受性并且对饮酒量有所不同的受体激动剂（Jiang等， 1991; Mitchell等人，2014年； Margolis等，2017）。这项研究提供了概念观察证明的证明电生理学可用于检测药物的差异。启用分析和比较大量分子，我建议在急性大脑中使用多电极外记录切片可大大增加我们可以从中记录的神经元的数量，从而增加方法。我们将研究对4种临床化合物的神经元反应（吗啡，芬太尼，羟考酮和丁丙诺啡）并使用Damgo，这是一种高度选择性的MOR激动剂，广泛用于临床前研究，作为附加比较器。奖励电路（VTA）中大脑区域中的大多数神经元，止痛电路（Habenula）和呼吸电路（前胞津综合体）在急性中发射大脑切片，这些大脑区域也高度表达了MOR。因此，我们可以使用更高的吞吐量方法，以测量这些大脑中每个大脑中不同动力学家的效力和效率区域，并比较对每个配体在单个神经元中的顺序应用的响应每个大脑区域。此外，我们将比较幼稚的动物和制作吗啡的结果依赖。通过提高我们对特定电路中单个神经元阿片类药物反应的理解这种基本的体内阿片类药物作用，这项工作将（1）证明使用这种方法的可行性快速表征化合物，（2）确定MOR功能的生物学变异，这将使更多有效用于涉及阿片类药物信号的临床问题的治疗性开发，包括使用阿片类药物疾病，饮酒障碍和对止痛药的耐受性。