Resolvin receptor signaling in trigeminal sensory neurons

三叉神经感觉神经元中的 Resolvin 受体信号传导

• 批准号: 10738862
• 负责人: Sangsu Bang
• 金额: $ 16.1万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10738862
• 关键词: ATAC-seq; Absence of pain sensation; Acute; Acute inflammatory pain; Address; Affect; Afferent Neurons; Agonist; American; Analgesics; Anatomy; Animal Model; Animals; Anti-Inflammatory Agents; Behavioral; Binding; Biological Assay; Bite Force; CMKLR1 gene; Capsaicin; Cells; Chronic; Chronic inflammatory pain; Clinical; Data; Databases; Detection; Development; Disease; Eating; Enzyme-Linked Immunosorbent Assay; Exhibits; Face; Female; Flow Cytometry; Functional disorder; Future; G-Protein-Coupled Receptors; Gene Expression Profile; Generations; Human; Immune; Impairment; Individual; Inflammation; Inflammation Mediators; Injections; Injury; Ion Channel; Macrophage; Measurement; Measures; Mechanics; Mediating; Methods; Modeling; Molecular; Mus; Nervous System; Neurogenic Inflammation; Neurons; Omega-3 Fatty Acids; Oral; Orofacial Pain; Pain; Pain management; Pathology; Pathway interactions; Pattern; Persistent pain; Persons; Production; Property; Receptor Signaling; Research; Resolution; Role; Saliva; Sampling; Secondary to; Series; Serum; Signal Pathway; Signal Transduction; Specificity; Spinal Ganglia; Structure of trigeminal ganglion; TNF gene; TRP channel; TRPV1 gene; Temporomandibular Joint; Temporomandibular Joint Disorders; Testing; Time; Tissues; Trigeminal Pain; Trigeminal System; United States National Institutes of Health; Vibrissae; Virus; Western Blotting; behavioral phenotyping; cell type; chronic pain; chronic pain management; clinical translation; conditioned place preference; craniofacial; drinking; effective therapy; experimental study; human tissue; inflammatory pain; inhibitor; innovation; insight; knock-down; lipid mediator; male; new therapeutic target; novel; novel therapeutics; orofacial; pain model; pain processing; prevent; receptor; sex; small hairpin RNA; spontaneous pain; tissue injury; translational impact

Project Summary/Abstract Pain management in craniofacial pathology is a major clinical challenge. Although many people suffer from chronic inflammatory pain caused by a temporomandibular joint disorder (TMJD) and other chronic oral and facial diseases, there is no effective treatment. Our previous study demonstrated that lipid mediators derived from omega-3 polyunsaturated fatty acids such as resolvin E1 (RvE1) were effective in inhibiting inflammatory pain by regulating TRPV1. However, it is unclear whether RvE1 can inhibit TMJ pain in the trigeminal nervous system, the role of its receptor ChemR23, and its relevance to clinical translation. We found from preliminary data that ChemR23 is co-expressed with TRPV1 using the ATAC-Seq database, RvE1 was decreased in the TMJ-CFA model using ELISA detection, and RvE1 can reduce TMJ pain using the biting force method. Based on this, we suggest that a RvE1/ChemR23 signaling pathway resolves TMJ pain by suppressing TRPV1 function. In Aim1, we predict that the RvE1/ChemR23 pathway will be downregulated during the TMJ-CFA model. To address our hypothesis, we will examine measuring of change of RvE1 level in TMJ-tissue, TGs, saliva, and serum in TMJ-CFA mice to confirm whether RvE1 can affect pain using the ELISA method. And we will test the ChemR23 expression level change and TRPV1 binding activity in TGs using western blot after the generation of the TMJ-CFA model. We will measure how much ChemR23 is expressed in TMJ-innervated TG in the TMJ-CFA model using flow cytometry. In Aim2, we will investigate the mechanism of pain suppression through ChemR23/TRPV1 through animal behavioral experiments. To know the RvE1 can block the TRPV1-induced pain in TG, we will cause acute trigeminal pain by injecting TRPV1 agonist capsaicin into the whisker pad. In addition, we will use unilateral CFA injection to the TMJ to test the hypothesis that RvE1 inhibits TMJ pain via ChemR23. To that end, we'll use local whisker pads or TMJ injections of the AAV1 virus to reduce ChemR23 expression in TG neurons. In Aim3, we will examine whether ChemR23 and TRPV1 mechanisms can be applied to human sensory neurons through Co-IP assay and real-time PCR using Human DRG and TG provided by Co-I. We will compare the differences in expression levels and Co-IP activity of DRGs and TGs. We propose a novel molecular mechanism for the resolution of tertiary pain where RvE1-induced ChemR23 activation can inhibit TRPV1 signaling. If completed, including future directions, this proposal will provide a new path for TMJ pain control. It will enable the development of new drugs aimed at TMJ and broaden our understanding of the disease. And this proposal can provide fundamental data for future research, such as investigating the role of additional resolvins and other TRP channels in human serum and chronic animal models.

项目总结/摘要 颅面病理学中的疼痛管理是一个主要的临床挑战。虽然很多人患有 由颞下颌关节紊乱病（TMJD）和其他慢性口腔和关节炎引起的慢性炎性疼痛， 面部疾病，没有有效的治疗方法。我们以前的研究表明，脂质介质衍生 从ω-3多不饱和脂肪酸如resolvin E1（RvE 1）有效抑制炎症 通过调节TRPV 1来缓解疼痛。然而，目前还不清楚RvE 1是否可以抑制三叉神经痛， 系统，其受体ChemR 23的作用，及其与临床翻译的相关性。 我们使用ATAC-Seq数据库从初步数据中发现ChemR 23与TRPV 1共表达， ELISA法检测到RvE 1在TMJ-CFA模型中降低，RvE 1可以减轻TMJ疼痛。 咬合力法基于此，我们认为RvE 1/ChemR 23信号通路通过以下途径解决TMJ疼痛： 抑制TRPV 1功能。 在Aim 1中，我们预测RvE 1/ChemR 23通路将在TMJ-CFA模型期间下调。到 为了解决我们的假设，我们将检查测量TMJ组织，TG，唾液中RvE 1水平的变化， TMJ-CFA小鼠的血清中进行测定，以使用ELISA方法确认RvE 1是否可以影响疼痛。我们将测试 使用蛋白质印迹法，在TGs中的ChemR 23表达水平变化和TRPV 1结合活性在TGs产生后， TMJ-CFA模型我们将测量在TMJ-CFA中TMJ神经支配的TG中有多少ChemR 23表达。 模型使用流式细胞术。 目的二是通过动物实验研究ChemR 23/TRPV 1的镇痛机制 行为实验RvE 1可以阻断TRPV 1诱导的TG疼痛， 通过将TRPV 1激动剂辣椒素注射到胡须垫中来治疗三叉神经疼痛。此外，我们将使用单边CFA 注射至TMJ以检验RvE 1通过ChemR 23抑制TMJ疼痛的假设。为此，我们将使用本地 晶须垫或颞下颌关节注射AAV 1病毒以减少TG神经元中的ChemR 23表达。 在Aim 3中，我们将研究ChemR 23和TRPV 1机制是否可以应用于人类感觉神经元 通过Co-IP测定和使用Co-I提供的人DRG和TG的实时PCR。我们将比较 DRG和TG的表达水平和Co-IP活性的差异。 我们提出了一种新的分子机制，用于解决三级疼痛，其中RvE 1诱导的ChemR 23 激活可抑制TRPV 1信号传导。如果完成，包括未来的方向，这一建议将提供一个新的 颞下颌关节疼痛控制路径。它将使针对颞下颌关节的新药的开发成为可能，并扩大我们的研究范围。 了解疾病。这一建议可以为未来的研究提供基础数据，例如， 研究人血清和慢性动物模型中额外的消退素和其他TRP通道的作用。