Adenylyl cyclase signaling in persistent pain

持续性疼痛中的腺苷酸环化酶信号传导

基本信息

批准号：
10418657
负责人：
DEREK C MOLLIVER
金额：
$ 45.99万
依托单位：
UNIVERSITY OF NEW ENGLAND
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10418657
关键词：
A kinase anchoring protein Acute Acute Pain Adenylate Cyclase Affect Afferent Neurons American Analgesics Animals Arthritis Automobile Driving Behavior assessment Behavioral Biochemical Brain CRISPR/Cas technology Cells Cessation of life Chronic Clinical Co-Immunoprecipitations Complex Coupled Cyclic AMP Cyclic AMP-Dependent Protein Kinases Dangerousness Development Diabetic Neuropathies Dinoprostone Electrophysiology (science)Family Family member G Protein-Coupled Receptor Signaling Gene Deletion Genes Goals Hyperalgesia Hypersensitivity Image Immunohistochemistry In Vitro Inflammatory Injury Intervention Intractable Pain Investigation Ion Channel Ion Channel Gating Knock-out Knockout Mice Lead Malignant Neoplasms Mass Spectrum Analysis Mechanics Mediating Modeling Molecular Mus Neurogenic Inflammation Neurons Neuropathy Neuropeptides Nociception Nociceptors Opioid Opioid Analgesics Opioid Receptor Pain Pain management Pathologic Peripheral Persistent pain Pharmaceutical Preparations Pharmacology Phosphoproteins Phosphorylation Play Postherpetic neuralgia Pre-Clinical Model Production Protein Isoforms Receptors, Adrenergic, alpha-2 Regulation Reporter Resistance Role Scaffolding Protein Second Messenger Systems Signal Pathway Signal Transduction Signaling Protein Spinal Cord Spinal Ganglia Sustainable Development Syndrome System TRPV1 gene Testing Western Blotting allodynia base chronic pain chronic pain management clinically significant effective intervention effective therapy experimental study improved in vivo innovation non-opioid analgesic novel pain chronification patch clamp prescription opioid receptor recruit response scaffold societal costs transmission process

项目摘要

Chronic pain affects more than 50 million Americans per year, resulting in extraordinary personal and societal costs. Adding to the dilemma, deaths involving prescription opiate analgesics have almost quadrupled in the last ten years. The clinical challenge of pain management is underscored by evidence that chronic pain is mechanistically distinct from acute pain, therefore a thorough understanding of the molecular and cellular mechanisms underlying the transition to chronic pain is fundamental to improving and expanding treatment options. Hyperalgesic priming is a compelling model of the transition to chronic pain in which an initial injury resolves, but leaves the animal in a primed state in which a second insult induces a greatly prolonged pain response. While previous studies have explored the development of sustained mechanical hypersensitivity, we have adapted this model to examine sustained hypersensitivity mediated by nociceptors expressing the heat-gated ion channel TRPV1, which can drive pain in a range of inflammatory and neuropathic conditions. Experiments proposed here will test the contribution to heat hyperalgesic priming of adenylyl cyclase isoform AC2, which has not been previously characterized in sensory neurons, and which is insensitive to inhibition by Gi/o-coupled receptors such as opioid receptors. Preliminary analysis indicates that AC2 is highly expressed in TRPV1-expressing neurons and required for the manifestation of heat hyperalgesic priming. Specific Aim 1 will examine the impact of AC2 gene deletion and pharmacological inhibition on behavioral nociceptive thresholds at baseline, in acute hyperalgesia and in the setting of hyperalgesic priming. Specific Aim 2 will determine whether AC2 and the downstream effector Epac are functionally coupled through AKAP family member scaffolding proteins by co-immunoprecipitation-mass spectrometry, and will determine the impact of deleting identified AKAP genes on Epac signaling and nociceptor function in vitro using a CRISPR/Cas9-based approach. Specific Aim 3 will determine the consequences for Epac function of AC2 and AKAP gene deletion through Epac-dependent PKC phospho- substrate profiling and behavioral assessment of heat hyperalgesia. This proposal will use innovative approaches to explore novel mechanisms contributing to the development of persistent hyperalgesia.

慢性疼痛每年影响超过5000万美国人，导致非凡个人和社会成本。增加困境，涉及处方鸦片的死亡在过去的十年中，镇痛药几乎近三倍。疼痛的临床挑战管理有证据表明慢性疼痛在机械上与急性不同疼痛，因此对分子和细胞机制的透彻理解向慢性疼痛的过渡对于改善和扩大治疗方案至关重要。高温启动是一种令人信服的过渡到慢性疼痛的模型伤害解决了，但使动物处于底漆状态，其中第二次侮辱会诱导A 大大延长疼痛反应。虽然先前的研究探讨了持续的机械性超敏反应，我们已经适应了该模型以检查持续的表达热门离子通道TRPV1的伤害感受器介导的超敏反应，该通道在一系列炎症和神经性疾病的范围内会疼痛。提出了实验这里将测试对腺苷环酶同工型AC2的热高致高致高致高变质启动的贡献，以前尚未在感觉神经元中表征，并且对 GI/O偶联受体（如阿片受体）抑制。初步分析表明 AC2在表达TRPV1的神经元中高度表达，并且表现为加热高效启动。具体目标1将检查AC2基因缺失的影响和基线时对行为伤害阈值的药理抑制作用，急性痛觉过敏和高温启动的环境。具体目标2将决定是否 AC2和下游效应子EPAC通过AKAP家族成员功能耦合通过共免疫沉淀物质光谱法脚手架蛋白质，将确定删除的影响鉴定出AKAP基因对EPAC信号传导和伤害感受器功能的影响使用基于CRISPR/CAS9的方法。具体目标3将决定 AC2和AKAP基因缺失的EPAC功能通过EPAC依赖性PKC磷酸热痛觉过敏的底物分析和行为评估。该建议将使用创新的方法来探索有助于发展的新型机制持续的痛觉过敏。