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：
10317189
负责人：
Amanda Helen Klein
金额：
$ 64.51万
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/10317189
关键词：
ATP sensitive potassium channel complex Acute Address Adenoviruses Adenylate Cyclase Affect Agonist Analgesics Attenuated Basic Science Cells Chronic Chronic inflammatory pain 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 Lead Mechanics Mediator of activation protein Mission Molecular Morphine Mus National Institute of Drug Abuse Nerve Fibers Nervous system structure Neural Conduction Neuraxis Neurons Nociceptors Opioid Opioid agonist Outcome Patients Peripheral Pharmacology 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 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 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.

项目概要拟议的研究与公共卫生相关，因为阿片类药物的使用在美国和人类中都很普遍。与容忍和退出相关的财务成本已达到危机水平。为了减少阿片类药物滥用和误用，我们的长期目标是确定导致这些临床问题的细胞内机制。该计划的目标拟议的研究是为了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慢性阿片类药物暴露期间的周围和中枢神经系统。已经做了大量的工作完成了对肥大腺苷酸环化酶活性和表达的矛盾现象的研究长期接触阿片类药物期间。中心假设是腺苷酸环化酶和下游活性增加介质降低 KATP 通道活性，导致神经元去极化并增加超敏反应和自发性疼痛。该提案的基本原理是，其完成将确定腺苷酸的关键细胞内靶标环化酶，包括钾通道，例如 ATP 敏感钾 (KATP) 通道，这将有助于我们进行分类改变神经元兴奋性的分子，可能在慢性阿片类药物暴露期间的超敏反应中发挥关键作用。鉴于神经系统中腺苷酸环化酶和抑制性 G 蛋白偶联信号传导的研究历史，令人惊讶的是，关于这些分子如何影响疼痛处理的神经生理学的基本问题仍然存在。我们的第一个假设是，背根神经节和脊髓中腺苷酸环化酶 1 的总体表达在慢性吗啡暴露。我们的第二个假设是腺苷酸环化酶 1 的上调，从而导致 cAMP，蛋白激酶 A 和 Epac 分子可降低体外 KATP 通道活性。我们的第三个假设是上调使用鞘内注射腺病毒病毒载体将背根神经节和脊髓中的 KATP 通道亚基上调腺苷酸环化酶后，可改善小鼠的机械超敏反应、活动能力和神经传导慢性阿片类药物暴露。这些方法应该被证明是相互补充的，并将提供最大的帮助有机会观察慢性阿片类药物暴露后神经系统发生的变化。我们计划解决这些假设是通过原位杂交、电生理学和钾通量测定的创新组合得出的体外和体内遗传方法。拟议的工作很重要，因为完成这些研究将确定腺苷酸环化酶和 KATP 通道功能之间的反比关系是否最终可能成为其基础并促进阿片类药物耐受和戒断期间临床上出现的疼痛信号传导。 KATP 通道呈现出未利用的状态开发治疗阿片类药物滥用和误用的药物的有趣目标。这些结果将产生积极的影响影响，因为它们将在过程中提供对这些信号转导途径的更多知识和理解阿片类药物暴露可能有助于为患者找到更好的当前镇痛疗法替代品。