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.

项目摘要/摘要 头面部病理中的疼痛管理是一项重大的临床挑战。尽管许多人遭受着 由颞下颌关节紊乱病(TMJD)和其他慢性口腔和其他疾病引起的慢性炎症性疼痛 面部疾病，目前还没有有效的治疗方法。我们之前的研究表明，脂质介质源于 来自omega-3的多不饱和脂肪酸，如解决素E1(RvE1)，可以有效地抑制炎症 通过调节TRPV1而产生疼痛。然而，目前尚不清楚RvE1是否能抑制三叉神经的TMJ痛。 系统，其受体ChemR23的作用，以及它与临床翻译的关系。 我们从初步数据中发现，利用ATAC-Seq数据库，ChemR23与TRPV1共表达， 在TMJ-CFA模型中，通过EL ISA检测，RvE1减少，并且RvE1可以通过 咬合力法。基于此，我们认为RvE1/ChemR23信号通路通过以下途径来解决TMJ疼痛 抑制TRPV1功能。 在Aim1中，我们预测在TMJ-CFA模型中RvE1/ChemR23通路将下调。至 为了解决我们的假设，我们将检查TMJ组织、TGS、唾液和 用ELISA法检测TMJ-CFA小鼠血清中RvE1对疼痛的影响。我们将测试 免疫印迹法检测TGS中ChemR23的表达水平及TRPV1结合活性 TMJ-CFA模式。我们将测量在TMJ-CFA中TMJ神经支配的TG中有多少ChemR23表达 模型采用流式细胞术。 在AIM2中，我们将通过动物来研究ChemR23/TRPV1抑制疼痛的机制 行为实验。要知道RvE1可以阻断TRPV1诱导的TG疼痛，我们将引起急性 三叉神经痛通过将TRPV1激动剂辣椒素注射到胡须垫内。此外，我们将使用单边终审法院 向TMJ注射RvE1，以验证RvE1通过ChemR23抑制TMJ疼痛的假设。为此，我们将使用本地 须垫或TMJ注射AAV1病毒以减少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Co-I提供的人DRG和TG的Co-IP分析和Real-Time PCR。我们将比较 DRGs和TGS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来解决三级疼痛，其中RvE1诱导了ChemR23 激活可抑制TRPV1信号转导。如果完成，包括未来的方向，这项建议将提供一个新的 TMJ疼痛控制路径。它将使针对TMJ的新药的开发成为可能，并扩大我们的 对疾病的了解。这一建议可以为未来的研究提供基础数据，如 研究额外的解析素和其他色氨酸通道在人血清和慢性动物模型中的作用。