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

基本信息

批准号：
10119457
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
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10119457
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 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 addiction base chronic pain compliance behavior design detector falls in silico 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 wound healing

项目摘要

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.

项目摘要炎症状态的主要特征之一是通常无害的刺激会导致疼痛。当前的疼痛的药物包括非甾体类抗炎药，旨在阻止前列腺素产生，皮质类固醇和阿片类药物。但是，副作用和某些情况成瘾与这些药物相关的潜力限制了它们的长期使用，尤其是在慢性炎症性疼痛期间。到开发新颖的，非成瘾性的镇痛药，迫切需要了解炎症产生痛苦感知的伤害感受器的变化。在此提案中，我们旨在提供在背根神经节（DRG）神经元中，Adaptin 2网格蛋白介导的内吞作用（AP2-- CME）是炎症引起的伤害感受器敏化的主要促进因子。我们以前有证明为响应蛋白激酶A（PKA）刺激，松弛的KNA通道通过来自DRG神经元膜的AP2-CME及其引起过度兴奋。此外，我们表明抑制AP2-CME阻止了PKA诱导的神经元过度兴奋性。现在的初步研究表明 DRG神经元内的AP2亚基AP2A2的体内敲低，大幅降低炎症性疼痛行为。在这里，我们将采用蛋白质化学，免疫组织化学的组合，电生理学，脊髓生理学，疼痛行为分析和一种新型的体内基因敲低方法为了测试AP2-CME是伤害感受器敏化的关键调节剂的假设。具体目的是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可缓解炎症性疼痛。该研究项目将揭示核心角色AP2-CME 发挥疼痛信号。