Fn14, non-canonical NF-kappaB and downstream signaling in neuropathic pain

Fn14、非典型 NF-kappaB 和神经性疼痛中的下游信号传导

• 批准号: 10175065
• 负责人: J. Marc Simard
• 金额: $ 37.31万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10175065
• 关键词: Animals; Antibodies; Apoptosis Promoter; Astrocytes; Brain; CXC Chemokines; Calcineurin; Cells; Centers for Disease Control and Prevention (U.S.); Chronic; Cytokine Receptors; Data; Development; Etiology; Exhibits; Exposure to; Fibroblast Growth Factor; Genes; Genetic; Genetic Transcription; Glial Fibrillary Acidic Protein; Glyburide; Goals; HIV; HIV Envelope Protein gp120; In Vitro; Inflammation; Inflammatory; Interleukin-6; Laboratories; Lead; Ligands; Light; Methods; Modeling; Molecular; Mus; NF-kappa B; Nerve; Nerve Growth Factors; Neuroglia; Neurons; Opiate Addiction; Opioid; Organ; Pain Research; Pain management; Pathogenesis; Pathologic; Pathway interactions; Patients; Peripheral nerve injury; Pharmaceutical Preparations; Pharmacology; Phenotype; Play; Prevention Guidelines; Proteins; Public Health; Role; Signal Transduction; Site; TNF gene; TNFRSF5 gene; Tissues; Up-Regulation; Work; addiction; allodynia; beta-Chemokines; chemokine; chemokine receptor; chromatin immunoprecipitation; chronic pain; cytokine; dorsal horn; evidence based guidelines; experimental study; in vivo; mu opioid receptors; neuroinflammation; new therapeutic target; non-opioid analgesic; novel; opioid epidemic; opioid overdose; pain behavior; pain model; painful neuropathy; prescription opioid; public health relevance

BACKGROUND: The ongoing opioid epidemic is driven in part by legitimate use of opioids prescribed for neuropathic pain. Targeting alternative pathways for pain control using non-addicting drugs is a major goal of neuropathic pain research. After peripheral nerve injury (PNI), cytokines and chemokines are upregulated centrally, where they contribute mechanistically to the pathogenesis of neuropathic pain. Reactive astrocytes in the dorsal horn exhibit a chronically activated, pro-inflammatory secretory (CAPS) phenotype characterized by secretion of numerous factors, including interleukin-6 (IL-6), chemokine C-C motif ligand 2 (CCL2), chemokine C-X-C motif ligand 1 (CXCL1) and nerve growth factor (NGF). The post-PNI astrocytic CAPS phenotype contributes to both inflammation and neuronal hyperactivation via neuronal chemokine receptors, leading to neuropathic pain. Canonical NF-κB signaling is known to play a crucial role, but surprisingly, despite its documented importance in numerous inflammatory conditions involving most organs including the brain, non- canonical NF-κB signaling via the p52:RelB heterodimer has not been identified previously in PNI. New work from our laboratory demonstrates the novel finding that non-canonical NF-κB signaling by the tumor necrosis factor-like weak inducer of apoptosis (TWEAK) / fibroblast growth factor-inducible 14 (Fn14) axis is prominent after PNI, especially in dorsal horn astrocytes, and may be responsible for transcriptional expression of Sur1- Trpm4 in these cells. Moreover, our preliminary data suggest that glial Sur1-Trpm4 plays a crucial role in regulating the expression of IL-6, CCL2 and CXCL1. Our central hypothesis is that, in dorsal horn astrocytes post-PNI, TWEAK-induced non-canonical NF-κB signaling is an upstream regulator of Sur1-Trpm4 expression, and that Sur1-Trpm4, in turn, regulates the expression of the downstream effectors, IL-6, CCL2 and CXCL1, which promote chronic neuroinflammation, neuronal hyperactivation and neuropathic pain. DESCRIPTION: This project has three mechanistic aims. In Aim 1, genetic (Tweak–/–, Fn14–/–, p100–/–, Abcc8–/–, either global or GFAP-specific or GFAP-&-site-specific) and pharmacological (anti-TWEAK antibody, glibenclamide) experiments will be carried out to determine the role of non-canonical NF-κB and Sur1 in GFAP-expressing glia in sciatic n. vs. dorsal horn in two neuropathic pain models: sciatic n. cuffing and sciatic n. exposure to HIV/gp120 protein. In Aim 2, we will expand upon in vivo and in vitro data from chromatin immunoprecipitation to establish the role of non-canonical NF-κB in the expression of functional Sur1-Trpm4 channels. In Aim 3, we will corroborate and expand upon in vivo and in vitro data to establish the role of Sur1- Trpm4 in regulating Ca2+/calcineurin-dependent transcription of IL-6, CCL2 and CXCL1. This project is the first to study non-canonical NF-κB in dorsal horn vs. sciatic n. glia in neuropathic pain, and will help identify novel druggable targets to ameliorate neuropathic pain using non-addictive drugs.

背景：当前阿片类药物流行的部分原因是合法使用阿片类药物 神经性疼痛。使用非成瘾药物控制疼痛的替代途径是一个主要目标 神经病理性疼痛研究。周围神经损伤（PNI）后，细胞因子和趋化因子上调 集中地，它们在机制上促进了神经性疼痛的发病机制。反应性星形胶质细胞 背角表现出慢性激活的促炎分泌（CAPS）表型，其特征是 分泌多种因子，包括白细胞介素 6 (IL-6)、趋化因子 C-C 基序配体 2 (CCL2)、趋化因子 C-X-C 基序配体 1 (CXCL1) 和神经生长因子 (NGF)。 PNI 后星形胶质细胞 CAPS 表型 通过神经元趋化因子受体导致炎症和神经元过度激活，从而导致 神经性疼痛。众所周知，规范的 NF-κB 信号传导发挥着至关重要的作用，但令人惊讶的是，尽管它 已记录在涉及大多数器官（包括大脑、非 之前在 PNI 中尚未发现通过 p52:RelB 异二聚体的经典 NF-κB 信号传导。新作品 我们实验室的新发现表明，肿瘤坏死的非典型 NF-κB 信号传导 类凋亡弱诱导剂 (TWEAK)/成纤维细胞生长因子诱导 14 (Fn14) 轴突出 PNI 后，特别是在背角星形胶质细胞中，可能负责 Sur1- 的转录表达 这些细胞中的 Trpm4。此外，我们的初步数据表明胶质细胞 Sur1-Trpm4 在 调节 IL-6、CCL2 和 CXCL1 的表达。我们的中心假设是，在背角星形胶质细胞中 PNI 后，TWEAK 诱导的非典型 NF-κB 信号传导是 Sur1-Trpm4 表达的上游调节因子， Sur1-Trpm4 反过来调节下游效应子 IL-6、CCL2 和 CXCL1 的表达， 促进慢性神经炎症、神经元过度激活和神经性疼痛。 描述：该项目具有三个机械目标。在目标 1 中，遗传（Tweak–/–、Fn14–/–、p100–/–、 Abcc8–/–，全局或 GFAP 特异性或 GFAP-&-位点特异性）和药理学（抗 TWEAK 抗体， 将进行格列本脲）实验以确定非典型 NF-κB 和 Sur1 在 坐骨神经胶质细胞表达 GFAP n。与背角在两种神经性疼痛模型中的比较：坐骨神经痛。袖套和坐骨神经 名词暴露于 HIV/gp120 蛋白。在目标 2 中，我们将扩展染色质的体内和体外数据 免疫沉淀以确定非典型 NF-κB 在功能性 Sur1-Trpm4 表达中的作用 渠道。在目标 3 中，我们将证实并扩展体内和体外数据，以确定 Sur1- 的作用 Trpm4 调节 IL-6、CCL2 和 CXCL1 的 Ca2+/钙调磷酸酶依赖性转录。这个项目是第一个 研究背角与坐骨神经中的非典型 NF-κB。神经胶质细胞在神经性疼痛中的作用，并将有助于识别新的 使用非成瘾药物缓解神经性疼痛的可药物靶标。