The role of the nociceptor endocytosis in inflammatory pain

伤害感受器内吞作用在炎性疼痛中的作用

基本信息

批准号：
10512357
负责人：
Arindam Bhattacharjee
金额：
$ 37.74万
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：
2022
资助国家：
美国
起止时间：
2022-06-15 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10512357
关键词：
14-3-3 Family Absence of pain sensation Acute Adaptor Protein Complex 2 Adaptor Protein Complex Subunits Adaptor Signaling Protein Adrenal Cortex Hormones Adverse effects Analgesics Arthralgia Arthritis Axon Binding Biochemical Biological Assay Cartilage Cell Fractionation Chronic Chronic inflammatory pain Clathrin Clinical Treatment Complex Cyclic AMP-Dependent Protein Kinases Data Dense Core Vesicle Development Drug Targeting Endocytosis Endocytosis Inhibition Fiber Future Genes Genetic Glucocorticoids Goals Histology Human Image Impairment Inflammation Inflammatory Injections Injury Intra-Articular Injections Joints Maintenance Measures Mediating Methods Modeling Molecular Mus Mutagenesis Neurons Nociception Nociceptors Outcome Pain Patients Peptides Peripheral Pharmaceutical Preparations Pharmacology Play Protein Family Protein Kinase C Proteins Rattus Reflex action Regulation Role Saline Signal Transduction Site Spinal Ganglia Steroid therapy Stress Structure Substance P TRPV1 gene Technology Testing Therapeutic Triamcinolone Work adverse drug reaction animal pain antagonist arthritic pain arthritis therapy chronic pain coping druggable target in vivo inflammatory pain inhibitor knock-down member microCT novel opioid overuse opioid use osteoarthritis pain pain behavior pain model pain reduction pain relief pain signal preservation prevent protein expression recruit residence response side effect trafficking treatment strategy vesicle transport

项目摘要

Project Summary The current pain killer crisis, fueled by the overuse of opioids to manage chronic inflammatory pain, urges the development of non-addictive pain relief medications. Local injection of drugs, at the site of injury, is one way to circumvent adverse drug reactions, including diversion potential. For example, injection of corticosteroids into arthritic joints is one of the mainstay therapeutic approaches to treat arthritic pain, circumventing opioid use. However, recent studies have put into question their efficacy and point to potential long-term deleterious effects of steroid therapy for arthritis. Thus, we require further identification of analgesic targets specifically at nociceptive peripheral terminals, i.e. local targets. We have recently identified the adaptin 2 alpha 2 subunit Ap2A2 and nociceptor endocytosis as promising targets for pain reduction in recoverable acute and chronic animal pain models. We showed that AP2A2 binds to KNa channels to cause their internalization and precipitate dorsal root ganglion (DRG) neuronal hyperexcitability after protein kinase A stimulation. The AP2A2 subunit localized to CGRP+/IB4- nociceptors. Genetically knocking down AP2A2 or locally inhibiting endocytosis with a lipidated decoy peptide provided prolonged reductions in pain behaviors in mouse and rats during inflammatory pain. We showed that nociceptor endocytosis was required for both the development and maintenance of inflammatory pain. Our exciting new preliminary data indicates that the AP2A2 subunit localizes to large dense core vesicles and are transported down axons where the AP2A2 subunit eventually finds residence at peripheral terminals. The first Aim of this study is to further understand the subcellular localization of AP2A2 and other members of the AP2 complex in peptidergic nociceptors. We will also study their expression during chronic pain. In the second Aim, we will test the contribution of AP2A2 and nociceptor endocytosis to pain in a non-recoverable inflammatory pain model: the monoiodoacetate-induced osteoarthritic pain model. We will use a genetic knockdown approach and intraarticular injection of our lipidated AP2 inhibitor peptide to study the consequences of nociceptor endocytosis inhibition on pain behaviors. Maintenance of inflammatory pain has been shown to be dependent upon protein kinase C (PKC) signaling. We had previously shown that Slack KNa channels constitutively associate with the 14-3-3 family of proteins in DRG neurons. Our preliminary data indicates that 14-3-3 zeta causes PKC-dependent Slack KNa channel internalization. In Aim 3 we will test the hypothesis that 14-3-3 proteins confer additional endocytotic motifs to KNa channels important for inflammatory signaling. We will use a novel lipidated 14-3-3 uncoupler peptide to study KNa channel trafficking in DRG neurons and pain behavior. Positive outcomes should establish the central role of nociceptor endocytosis to inflammatory pain and reveal druggable targets.

项目概要当前的止痛药危机是由于过度使用阿片类药物来控制慢性炎症疼痛而加剧的，敦促开发非成瘾性止痛药物。在受伤部位局部注射药物是一种方法规避药物不良反应，包括转移潜力。例如，注射皮质类固醇进入关节炎关节是治疗关节炎疼痛的主要治疗方法之一，避免阿片类药物使用。然而，最近的研究对其功效提出了质疑，并指出了潜在的长期有害作用。类固醇治疗关节炎的效果。因此，我们需要进一步确定镇痛目标，特别是伤害性外围终端，即局部目标。我们最近发现了适应素 2 α 2 亚基 Ap2A2 和伤害感受器内吞作用是减轻可恢复的急性和慢性疼痛的有希望的目标动物疼痛模型。我们发现 AP2A2 与 KNa 通道结合以引起其内化和蛋白激酶 A 刺激后促进背根神经节 (DRG) 神经元过度兴奋。 AP2A2 亚基定位于 CGRP+/IB4- 伤害感受器。基因敲低 AP2A2 或局部抑制脂质化诱饵肽的内吞作用可长期减少小鼠和大鼠的疼痛行为炎性疼痛期间。我们表明，伤害感受器内吞作用对于发育和发育都是必需的。维持炎性疼痛。我们令人兴奋的新初步数据表明 AP2A2 亚基定位于大而致密的核心囊泡，并沿着轴突运输，最终到达 AP2A2 亚基在外围终端找到住处。本研究的第一个目的是进一步了解亚细胞 AP2A2 和 AP2 复合物的其他成员在肽能伤害感受器中的定位。我们也会学习他们在慢性疼痛期间的表达。在第二个Aim中，我们将测试AP2A2和伤害感受器的贡献不可恢复的炎性疼痛模型中的内吞作用对疼痛的影响：单碘乙酸诱导的骨关节炎疼痛模型。我们将使用基因敲低方法和关节内注射我们的脂质化 AP2 抑制剂肽来研究伤害感受器内吞作用抑制对疼痛行为的影响。维护炎症性疼痛已被证明依赖于蛋白激酶 C (PKC) 信号传导。我们之前有过结果表明，Slack KNa 通道与 DRG 神经元中的 14-3-3 蛋白家族组成性相关。我们的初步数据表明 14-3-3 zeta 导致 PKC 依赖性 Slack KNa 通道内化。目标 3 我们将测试以下假设：14-3-3 蛋白向 KNa 通道赋予额外的内吞基序，这一点很重要用于炎症信号传导。我们将使用新型脂化 14-3-3 解偶联肽来研究 KNa 通道 DRG 神经元的贩运和疼痛行为。积极的结果应确立伤害感受器的核心作用内吞作用引起炎症疼痛并揭示可药物靶点。