In Vivo Implications of Agonist Selective Activation of Delta Opioid Receptor

Delta 阿片受体激动剂选择性激活的体内影响

基本信息

批准号：
8248182
负责人：
Amynah Amir Ali Pradhan
金额：
$ 15.85万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-01 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8248182
关键词：
Absence of pain sensation Accounting Acute Adult Adverse effects Affect Agonist Analgesics Animals Anxiety Arrestins Behavior Behavioral Binding Biological Assay California Chronic Chronic Disease Clinical Collaborations Complex Convulsions Core Facility Coupling DNA Microarray Chip Development Educational process of instructing Electrophysiology (science)Emotional Faculty Fentanyl G-Protein-Coupled Receptors GTP-Binding Proteins Gene Activation Gene Expression Genes Goals In Vitro Institution Ion Channel Knock-out Knockout Mice Laboratories Learning Ligands Ligation Los Angeles Mediating Mediator of activation protein Mental Depression Mentors Modeling Morphine Mus Mutant Strains Mice Neurons Opioid Opioid Receptor Pain Pharmacology Pharmacotherapy Phase Phosphoproteins Positioning Attribute Postdoctoral Fellow Primary Cell Cultures Property Proteins RNA Interference Receptor Activation Receptor Down-Regulation Receptor Inhibition Receptor Signaling Research Research Personnel Role Seizures Signal Transduction Signaling Protein Site Spinal Ganglia Structure Technical Expertise Techniques Therapeutic Time Universities Validation Work addiction arrestin 1 arrestin 2 brain tissue cell type chronic pain delta opioid receptor desensitization experience in vivo in vivo Model inflammatory pain inhibitor/antagonist interest knowledge base novel public health relevance receptor receptor binding receptor internalization response skills trafficking

项目摘要

DESCRIPTION (provided by applicant): I am currently a senior level postdoctoral fellow with a primary research focus in opioid receptor pharmacology. I intend to transition to a junior faculty position in the next two years, and my current mentored position has been structured to accomplish this goal. I have a wide-range of experience with in vivo models of pain and addiction, as well as extensive experience with techniques to assess receptor binding, function, and localization ex vivo. I am currently working in the laboratory of Dr. Christopher Evans, a prominent opioid researcher, at the University of California Los Angeles (UCLA). During my time at UCLA, I will enhance my skill set by learning new techniques in primary cell culture, electrophysiology, and arrays. My mentor and I have also planned several strategies to facilitate my transition to a faculty position, including budgetary responsibility, teaching, and course work. UCLA is a renowned institution with a number of high profile researchers, and during this mentored phase I will develop important collaborations that I hope to sustain into my independence. In addition, the university also has several state-of-the-art core facilities through which I will enhance my research opportunities. My research focus is on the in vivo consequences of ligand directed signaling at the delta (4) opioid receptor. Agonists for this receptor are being developed for clinical use, as activation of 4 receptors relieves pain, and reduces anxiety and depression. However, a major limitation to the clinical use of these compounds is that a subset of 4 agonists also produce convulsions, the mechanism of which is unknown. This agonist-selective behavior could be due to differential trafficking and signaling of the 4 receptor, and my recent work shows that the internalizing agonist, SNC80, produces convulsions which are not observed with the non-internalizing agonist, ARM390. 2-arrestins are major mediators of receptor internalization, and also mediate subsequent receptor signaling. One of the aims of this proposal is to determine the role of 2-arrestins in SNC80 and ARM390-induced behaviors, using 2-arrestin 1 and 2 knockout mice. In addition, to characterize the different signaling mechanisms regulating this functional selectivity, the distinct signaling complexes formed by SNC80 and ARM390 will be determined using proximity ligation assays and phosphoprotein arrays. Furthermore, I have found that repeated use of these agonists in an inflammatory pain model also results in differential tolerance. SNC80 produces receptor down regulation and generalized tolerance to all agonist-induced effects. In contrast, ARM390-tolerant animals show intact 4 receptors, and analgesic tolerance only - the mechanism of which is unknown. A further aim of this application is to characterize this differential tolerance, by looking for changes in receptor-ion channel coupling within the dorsal root ganglia following chronic use. I will also examine the role of 2-arrestins in these two types of tolerance, and explore possible mechanisms underlying these differences using DNA microarrays. This work has important therapeutic implications, and will enhance our understanding of in vivo opioid receptor trafficking and signaling. PUBLIC HEALTH RELEVANCE: Delta opioid receptor activation produces pain-relief, and reduces anxiety and depression; and for this reason agonist to this receptor are being developed for clinical use. However, a deterrent to the development of these compounds is that some delta agonists also produce convulsions, and at the moment there is no way to predict whether a novel agonist will have this property. The goal of this proposal is to determine the mechanism that accounts for this difference between agonists that bind to the same receptor. Ultimately, this work would increase the potential for the delta opioid receptor as a novel drug therapy.

描述（由申请人提供）：我目前是一名高级博士后研究员，主要研究方向为阿片受体药理学。我打算在未来两年内过渡到初级教员职位，而我目前的指导职位就是为了实现这一目标而设计的。我在疼痛和成瘾的体内模型方面拥有广泛的经验，并且在离体评估受体结合、功能和定位的技术方面也拥有丰富的经验。我目前在加州大学洛杉矶分校 (UCLA) 著名阿片类药物研究员 Christopher Evans 博士的实验室工作。在加州大学洛杉矶分校期间，我将通过学习原代细胞培养、电生理学和阵列方面的新技术来增强我的技能。我和我的导师还计划了几项策略来促进我向教员职位的过渡，包括预算责任、教学和课程工作。加州大学洛杉矶分校是一所著名的机构，拥有许多知名研究人员，在这个指导阶段，我将开展重要的合作，我希望将这些合作维持到我的独立性。此外，大学还拥有几个最先进的核心设施，通过这些设施我将增加我的研究机会。我的研究重点是 δ (4) 阿片受体配体定向信号传导的体内后果。该受体的激动剂正在开发用于临床，因为 4 受体的激活可以缓解疼痛，并减少焦虑和抑郁。然而，这些化合物临床使用的一个主要限制是 4 种激动剂中的一部分也会产生惊厥，其机制尚不清楚。这种激动剂选择性行为可能是由于 4 受体的不同运输和信号传导所致，我最近的研究表明，内化激动剂 SNC80 会产生用非内化激动剂 ARM390 未观察到的惊厥。 2-抑制蛋白是受体内化的主要介质，并且还介导随后的受体信号传导。该提案的目的之一是使用 2-arrestin 1 和 2 敲除小鼠确定 2-arrestin 在 SNC80 和 ARM390 诱导的行为中的作用。此外，为了表征调节这种功能选择性的不同信号传导机制，将使用邻近连接测定和磷蛋白阵列来确定由 SNC80 和 ARM390 形成的不同信号传导复合物。此外，我发现在炎性疼痛模型中重复使用这些激动剂也会导致不同的耐受性。 SNC80 产生受体下调和对所有激动剂诱导效应的普遍耐受。相比之下，ARM390耐受的动物表现出完整的4个受体，并且仅具有镇痛耐受性——其机制尚不清楚。本申请的另一个目的是通过寻找长期使用后背根神经节内受体-离子通道耦合的变化来表征这种差异耐受性。我还将研究 2-arrestins 在这两种耐受性中的作用，并使用 DNA 微阵列探索这些差异背后的可能机制。这项工作具有重要的治疗意义，并将增强我们对体内阿片受体转运和信号传导的理解。公共卫生相关性：Delta 阿片受体激活可缓解疼痛，并减少焦虑和抑郁；因此，正在开发该受体的激动剂用于临床。然而，阻碍这些化合物发展的一个因素是，一些δ激动剂也会产生惊厥，目前无法预测新型激动剂是否具有这种特性。该提案的目的是确定导致与相同受体结合的激动剂之间存在这种差异的机制。最终，这项工作将增加 δ 阿片受体作为新型药物疗法的潜力。