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）：我目前是高级博士后研究员，在阿片类受体药理学方面主要研究重点。我打算在未来两年内过渡到初级教师职位，而我目前的指导职位已构成实现这一目标。我在体内疼痛和成瘾模型以及在评估受体结合，功能和本地化的技术方面具有广泛的经验。我目前正在加利福尼亚大学洛杉矶分校（UCLA）的著名阿片类研究人员克里斯托弗·埃文斯（Christopher Evans）博士工作。在UCLA期间，我将通过学习原代细胞培养，电生理学和阵列中的新技术来提高自己的技能。我和我的导师还计划了几种策略，以促进我过渡到教师职位，包括预算责任，教学和课程工作。加州大学洛杉矶分校（UCLA）是一家拥有许多知名研究人员的著名机构，在这一指导阶段，我将建立重要的合作，希望能够维持自己的独立性。此外，该大学还拥有几个最新的核心设施，通过这些设施，我将增加我的研究机会。我的研究重点是在三角洲（4）阿片受体中配体的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用途，因为4种受体的激活可缓解疼痛，并减轻焦虑和抑郁。但是，这些化合物的临床使用的主要局限性是4个激动剂的子集也会产生抽搐，其机制尚不清楚。这种激动剂选择性的行为可能是由于4受体的差异运输和信号传导，我最近的工作表明，内部化激动剂SNC80会产生抽搐，而非内部化激动剂ARM390未观察到这些抽搐。 2-次素是受体内在化的主要介体，并且还介导了随后的受体信号传导。该提案的目的之一是使用2-甲蛋白1和2敲除小鼠确定2-次素在SNC80和ARM390诱导的行为中的作用。此外，为了表征调节该功能选择性的不同信号传导机制，将使用接近结扎测定和磷蛋白阵列确定由SNC80和ARM390形成的不同信号传导复合物。此外，我发现在炎症疼痛模型中反复使用这些激动剂也会导致差异。 SNC80可产生受体的调节，并对所有激动剂诱导的作用产生普遍的耐受性。相比之下，ARM390耐受性动物显示完整的4受体和镇痛耐受性 - 其机制尚不清楚。该应用程序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-次素在这两种类型的耐受性中的作用，并探索使用DNA微阵列差异的可能机制。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运输和信号传导的理解。公共卫生相关性：三角洲阿片受体激活会产生疼痛浮雕，并减少焦虑和抑郁；因此，正在开发该受体的激动剂供临床使用。但是，对这些化合物的发展的威慑是一些三角洲激动剂也会产生抽搐，目前无法预测新型激动剂是否会拥有这种特性。该提案的目的是确定与同一受体结合的激动剂之间这种差异的机制。最终，这项工作将增加Delta阿片受体作为一种新型药物疗法的潜力。