The role of the AP2 adaptor complex in inflammatory pain

AP2 接头复合物在炎性疼痛中的作用

• 批准号: 9920269
• 负责人: Arindam Bhattacharjee
• 金额: $ 6.34万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9920269
• 关键词: Absence of pain sensation; Action Potentials; Acute; Acute Pain; Adaptor Protein Complex 2; Adaptor Protein Complex Subunits; Adaptor Signaling Protein; Adrenal Cortex Hormones; Adult; Affect; Analgesics; Animals; Area; Attention; Attenuated; Behavior; Biological Assay; Chronic inflammatory pain; Clathrin; Computer Simulation; Cyclic AMP-Dependent Protein Kinases; Data; Elderly; Electrophysiology (science); Endocytosis; Evolution; Formalin; Future; Genes; Goals; Hip Fractures; Hormones; Hypotension; Immunohistochemistry; Inflammation; Inflammation Mediators; Inflammatory; Injections; Injury; Ion Channel; Knock-out; Knockout Mice; Lead; Mechanics; Mediating; Membrane; Methods; Mus; Neurons; Nociception; Nociceptors; Non-Steroidal Anti-Inflammatory Agents; Opioid; Outcome; Pain; Pain Measurement; Pain management; Patch-Clamp Techniques; Persistent pain; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phosphotransferases; Physiology; Plasmids; Play; Potassium; Process; Production; Property; Prostaglandin Production; Protein Chemistry; Publishing; Research Project Grants; Role; Signal Transduction; Sleep Apnea Syndromes; Slice; Sodium; Spinal Cord; Spinal Ganglia; Spinal nerve structure; Stimulus; Structure; Synaptic Transmission; TFAP2A gene; Techniques; Testing; Tissues; Transgenic Organisms; United States; Wound Healing; addiction; base; chronic pain; compliance behavior; design; detector; falls; in vivo; inflammatory pain; injured; knock-down; minimally invasive; neuronal excitability; neurotransmission; novel; pain behavior; pain model; pain perception; pain processing; pain relief; pain signal; prevent; protein complex; receptor; repaired; response; screening; side effect; small hairpin RNA; treatment strategy

Project Summary One of the cardinal features of inflammatory states is that normally innocuous stimuli produce pain. Current pain-relieving drugs include nonsteroidal anti-inflammatory drugs, which are aimed at the interdiction of prostaglandin production, corticosteroids and opioids. However, the side effects and some cases addiction potential associated with these drugs limit their long-term use especially during chronic inflammatory pain. To develop novel, non-addictive analgesics, there remains an urgent need to understand how inflammation produces the change in nociceptor firing that underlies pain perception. In this proposal, we aim to provide proof of principal that in dorsal root ganglion (DRG) neurons, adaptin 2 clathrin-mediated endocytosis (AP2- CME) is a principal facilitator of inflammatory-induced nociceptor sensitization. We have previously demonstrated that in response to protein kinase A (PKA) stimulation, Slack KNa channels are internalized via AP2-CME from DRG neuronal membranes and this caused hyperexcitability. Furthermore we showed that inhibiting AP2-CME prevented PKA-induced neuronal hyperexcitability. Preliminary studies now indicate that in vivo knockdown of the AP2 alpha subunit AP2A2 specifically within DRG neurons, substantially reduces inflammatory pain behavior. Here, we will apply a combination of protein chemistry, immunohistochemistry, electrophysiology, spinal cord physiology, pain behavior assays and a novel in vivo gene knockdown approach to test the hypothesis that AP2-CME is a key regulator of nociceptor sensitization. The specific aims are 1) to determine whether AP2-CME controls basal excitability and neurotransmission 2) To demonstrate that reducing AP2-CME mitigates inflammatory pain. This research project will reveal the central role AP2-CME plays in pain signaling.

项目概要 炎症状态的主要特征之一是通常无害的刺激会产生疼痛。当前的 止痛药包括非甾体类抗炎药，其目的是阻止疼痛 前列腺素的产生、皮质类固醇和阿片类药物。然而，副作用和某些情况下成瘾 与这些药物相关的潜力限制了它​​们的长期使用，尤其是在慢性炎症疼痛期间。到 开发新型、非成瘾性镇痛药，仍然迫切需要了解炎症是如何产生的 产生疼痛感受基础的伤害感受器放电的变化。在本提案中，我们的目标是提供 原理证明，在背根神经节 (DRG) 神经元中，适应素 2 网格蛋白介导的内吞作用 (AP2- CME）是炎症诱导的伤害感受器敏化的主要促进者。我们之前有过 证明，响应蛋白激酶 A (PKA) 刺激，Slack KNa 通道通过以下途径内化： AP2-CME 来自 DRG 神经元膜，这导致过度兴奋。此外我们还表明 抑制 AP2-CME 可防止 PKA 诱导的神经元过度兴奋。目前初步研究表明，在 体内敲低 AP2 α 亚基 AP2A2，特别是在 DRG 神经元内，显着降低 炎症性疼痛行为。在这里，我们将结合蛋白质化学、免疫组织化学、 电生理学、脊髓生理学、疼痛行为测定和新型体内基因敲除方法 检验 AP2-CME 是伤害感受器敏化的关键调节因子的假设。具体目标是 1) 确定 AP2-CME 是否控制基础兴奋性和神经传递 2) 证明 减少 AP2-CME 可减轻炎性疼痛。该研究项目将揭示 AP2-CME 的核心作用 参与疼痛信号传导。