KATP Channels as Downstream targets of adenylyl cyclases during opioid tolerance and withdrawal

KATP 通道作为阿片类药物耐受和戒断期间腺苷酸环化酶的下游靶标

基本信息

批准号：
10618258
负责人：
Amanda Helen Klein
金额：
$ 54.46万
依托单位：
UNIVERSITY OF MINNESOTA
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：
2021-08-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10618258
关键词：
ATP sensitive potassium channel complex Absence of pain sensation Acute Address Adenoviruses Adenylate Cyclase Affect Agonist Analgesics Attenuated Basic Science Cells Central Nervous System Chronic Chronic inflammatory pain Classification Clinical Coupled Cyclic AMP Cyclic AMP-Dependent Protein Kinases Data Development Down-Regulation Drug Targeting Drug usage Electrophysiology (science)Exposure to Financial cost Future G-Protein-Coupled Receptors GTP-Binding Proteins Goals Human Hypersensitivity Hypertrophy In Situ Hybridization In Vitro Incidence Intervention Intrathecal Injections Knowledge Mechanics Mediator Mission Molecular Morphine Mus National Institute of Drug Abuse Nerve Fibers Nervous System Neural Conduction Neurons Nociceptors Opioid Opioid agonist Outcome Patients Peripheral Nervous System Play Potassium Potassium Channel Protein Isoforms Public Health Recording of previous events Research Rodent Model Signal Transduction Signal Transduction Pathway Spinal Cord Spinal Ganglia Therapeutic United States Up-Regulation Viral Vector Withdrawal Withdrawal Symptom Work adenylyl cyclase 1 antagonist base chronic pain relief combat drug development genetic approach genetic predictors improved in vitro Assay in vitro activity in vivo innovation knock-down mu opioid receptors neurobiological mechanism neuronal excitability neurophysiology novel novel therapeutics opiate tolerance opioid abuse opioid exposure opioid misuse opioid use opioid withdrawal pain patient pain processing pain signal pharmacologic prescription opioid prevent response sciatic nerve spontaneous pain

项目摘要

Project Summary The proposed research is relevant to public health because opioid use is prevalent in the United States and the human and financial costs associated with tolerance and withdrawal are at crisis levels. In order to reduce opioid abuse and misuse, our long-term goal is to determine the intracellular mechanisms to lead to these clinical problems. The objective of the proposed research is to understand the molecular involvement of adenylyl cyclase signaling and potassium channels in the peripheral and central nervous system during chronic opioid exposure using rodent models. A great deal of work has been done investigating the paradoxical phenomena of hypertrophied adenylyl cyclase activity and expression that occurs during chronic opioid exposure. The central hypothesis is that increased activity adenylyl cyclase and downstream mediators decrease KATP channel activity, leading to neuronal depolarization and increased hypersensitivity and spontaneous pain. The rationale of this proposal is that its completion will identify key intracellular targets of adenylyl cyclases, including potassium channels such as ATP-sensitive potassium (KATP) channels, which will help us to classify molecules that alter neuronal excitability and may play a key role in hypersensitivity during chronic opioid exposure. Given the history of research into adenylyl cyclase and inhibitory G-protein coupled signaling in the nervous system, it is surprising that fundamental questions still exist as to how these molecules affect neurophysiology of pain processing. Our first hypothesis is that overall expression of adenylyl cyclase 1 in the dorsal root ganglia and spinal cord increases after chronic morphine exposure. Our second hypothesis is that upregulation of adenylyl cyclase 1, and consequently cAMP, protein kinase A, and Epac molecules decrease KATP channel activity in vitro. Our third hypothesis is that upregulation of KATP channel subunits in the dorsal root ganglia and spinal cord using intrathecal injection of adenovirus viral vectors will improve mechanical hypersensitivity, mobility, and nerve conduction in mice after upregulated adenylyl cyclase during chronic opioid exposure. These approaches should prove to be complementary to one another and will provide the greatest opportunity to observe changes that occur in the nervous system after chronic opioid exposure. We plan on addressing these hypotheses through an innovative combination of in situ hybridization, electrophysiology, and potassium flux assays in vitro, and genetic approaches in vivo. The proposed work is important because completion of these studies will determine if the inverse relationship between adenylyl cyclase and KATP channel functionality could ultimately underlie and promote pain signaling seen clinically during opioid tolerance and withdrawal. KATP channels present an unutilized and interesting target for the development of drugs to treat opioid abuse and misuse. These results will have a positive impact because they will provide an increased knowledge and understanding of these signal transduction pathways during opioid exposure may assist in the mission to find better alternatives to current analgesic therapies for patients.

项目摘要拟议的研究与公共卫生有关，因为在美国和人类和与容忍和退出相关的财务成本处于危机水平。为了减少阿片类药物滥用和滥用，我们的长期目标是确定导致这些临床问题的细胞内机制。目的拟议的研究是了解腺苷酸环化酶信号传导和钾通道的分子参与使用啮齿动物模型在慢性阿片类药物暴露期间的外围和中枢神经系统。大量工作已经研究了肥厚的腺苷环酶活性和发生的表达的悖论现象在慢性阿片类药物暴露期间。中心假设是活性增加了腺苷酸环化酶和下游介体降低了KATP通道活性，导致神经元去极化并增加了超敏反应和增加自发疼痛。该提议的理由是，其完成将确定adenylyl的关键细胞内目标循环酶，包括钾通道，例如ATP敏感钾（KATP）通道，这将帮助我们对改变神经元兴奋性并在慢性阿片类药物暴露期间的超敏反应中起关键作用的分子。鉴于神经系统中对腺苷酸环化酶和抑制性G蛋白偶联信号的研究历史，这是令人惊讶的是，对于这些分子如何影响疼痛处理的神经生理学仍存在基本问题。我们的第一个假设是，在背根神经节中腺苷酸环化酶1的总体表达在慢性吗啡暴露。我们的第二个假设是，腺苷环化酶1的上调，因此是cAMP，蛋白激酶A和EPAC分子在体外降低KATP通道活性。我们的第三个假设是使用腺病毒病毒载体的脑膜根神经节和脊髓中的KATP通道亚基将在上调腺苷环酶后，改善小鼠的机械性超敏反应，迁移率和神经传导慢性阿片类药物暴露。这些方法应证明是彼此互补的，将提供最大的观察慢性阿片类药物暴露后神经系统中发生的变化的机会。我们计划解决这些假设通过原位杂交，电生理学和钾通量测定的创新组合体内体外和遗传方法。拟议的工作很重要，因为这些研究的完成将确定腺苷酸环化酶和KATP通道功能之间的逆关系最终可能是基础并促进阿片类药物耐受性和戒断期间临床观察到的疼痛信号。 KATP频道提出了一个未利用的以及开发药物来治疗阿片类药物滥用和滥用的有趣目标。这些结果将具有积极的影响是因为它们会在期间对这些信号转导途径的知识和理解增加阿片类药物的暴露可能有助于为患者找到当前镇痛药疗法的更好替代方法。