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.

项目概要/摘要 颅面病理学中的疼痛管理是一项重大的临床挑战。虽然很多人都遭受着 由颞下颌关节紊乱 (TMJD) 和其他慢性口腔和口腔疾病引起的慢性炎性疼痛 面部疾病目前尚无有效治疗方法。我们之前的研究表明，脂质介质衍生 来自 omega-3 多不饱和脂肪酸，例如 Resolvin E1 (RvE1)，可有效抑制炎症 通过调节 TRPV1 来产生疼痛。然而，目前尚不清楚RvE1是否可以抑制三叉神经的TMJ疼痛。 系统、其受体 ChemR23 的作用及其与临床转化的相关性。 我们利用ATAC-Seq数据库从初步数据发现ChemR23与TRPV1共表达， ELISA检测TMJ-CFA模型中RvE1降低，RvE1可以减轻TMJ疼痛 咬力法。基于此，我们建议 RvE1/ChemR23 信号通路通过以下方式解决 TMJ 疼痛： 抑制 TRPV1 功能。 在 Aim1 中，我们预测 RvE1/ChemR23 通路在 TMJ-CFA 模型中将会下调。到 为了解决我们的假设，我们将检查 TMJ 组织、TG、唾液和 使用 TMJ-CFA 小鼠血清，使用 ELISA 方法确认 RvE1 是否可以影响疼痛。我们将测试 使用蛋白质印迹法生成 TG 后 ChemR23 表达水平变化和 TRPV1 结合活性 TMJ-CFA 模型。我们将测量 TMJ-CFA 中 TMJ 神经支配的 TG 中 ChemR23 的表达量 使用流式细胞仪建立模型。 在Aim2中，我们将通过动物研究ChemR23/TRPV1抑制疼痛的机制 行为实验。要知道RvE1可以阻断TRPV1引起的TG疼痛，我们将引起急性疼痛 通过将 TRPV1 激动剂辣椒素注射到胡须垫中来缓解三叉神经疼痛。另外，我们会使用单边CFA 注射到 TMJ 以测试 RvE1 通过 ChemR23 抑制 TMJ 疼痛的假设。为此，我们将使用本地 胡须垫或 TMJ 注射 AAV1 病毒可减少 TG 神经元中 ChemR23 的表达。 在Aim3中，我们将研究ChemR23和TRPV1机制是否可以应用于人类感觉神经元 通过使用 Co-I 提供的人类 DRG 和 TG 进行 Co-IP 测定和实时 PCR。我们将比较 DRGs 和 TGs 表达水平和 Co-IP 活性的差异。 我们提出了一种解决三级疼痛的新分子机制，其中 RvE1 诱导的 ChemR23 激活可以抑制 TRPV1 信号传导。如果完成，包括未来的方向，该提案将提供一个新的 TMJ 疼痛控制的路径。它将促进针对 TMJ 的新药物的开发，并扩大我们的研究范围。 对疾病的了解。并且该提案可以为未来的研究提供基础数据，例如 研究额外的消解素和其他 TRP 通道在人血清和慢性动物模型中的作用。