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Peripheral Receptor Mechanisms in Orofacial Muscle Pain

口面部肌肉疼痛的外周受体机制

基本信息

批准号：
10361180
负责人：
JIN Y Ro
金额：
$ 36.33万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10361180
关键词：
Achievement Attenuated Binding Cations Chemosensitization Chimeric Proteins Chronic Clustered Regularly Interspaced Short Palindromic Repeats DNA DNA Binding DNA Maintenance DNA Methylation DNMT3a Data Development Environmental Risk Factor Epigenetic Process Funding Fusion Protein Expression Gene Cluster Gene Expression Alteration Genes Genetic Transcription Hyperalgesia Individual Inflammation Inflammatory Island Lead Link Masseter Muscle Mechanics Mediating Membrane Methylation Molecular Muscle Myalgia Myositis N-Methylaspartate Nociception Nociceptors Orofacial Pain Outcome Oxidative Stress Pain Pathologic Peripheral Play Production Promoter Regions Property Psychophysics Reactive Oxygen Species Regulation Rodent Model Role Sensory Ganglia Signal Pathway Signal Transduction Stress Structure of trigeminal ganglion TRPA channel TRPV1 gene Technology Testing Tissues Up-Regulation base chronic painful condition craniofacial demethylation design epigenome editing genome wide methylation human disease inflammatory pain innovation novel orofacial prevent promoter protein complex prototype receptor response therapeutic development tool

项目摘要

PROJECT SUMMARY DNA methylation, an epigenetic factor, plays an important role in regulating gene expression and alterations in DNA methylation is a feature associated with a number of human diseases. Inflammation and environmental factors such as psychophysical stress induces demethylation of pro-nociceptive genes leading to their aberrant expression. The objective of this renewal application is to investigate how muscle inflammation remotely regulates DNA methylation of multiple pro-nociceptive genes in trigeminal ganglia (TG) that have been implicated in pain and hyperalgesia. Our central hypothesis is that masseter muscle inflammation results in reduced methylation of pro-nociceptive genes in TG leading to their aberrant expression, which contributes to the development of pain and mechanical hyperalgesia. We further hypothesize that psychophysical stress potentiates these effects via the excess production of reactive oxygen species (ROS) within TG, which regulate DNA methylation. In Aim 1, we will determine the role of DNA methylation in inflammatory pain responses. Specifically, we will examine whether increased DNA methylation via DNMTs at the promoter region of individual pro-nociceptive genes, prevents inflammatory pain and hyperalgesia. We have confirmed that pro-nociceptive genes such as TRPV1, TRPA1, P2X3 and PIEZO2 contain CG islands that bind DNMTs and that the inhibition of DNMT activities increased their expression in TG. In order to determine the role of DNA methylation in individual genes, we designed and validated a novel DNMT fusion protein complex that targets the promoter region of a specific gene, using the CRISPR-dCAS9 technology. We expect that the expression of the fusion protein within TG will prevent the upregulation of the target gene and reveal the relative contribution of DNA methylation for a specific gene in pain and hyperalgesia under a myositis condition. In Aim 2, we will investigate the role of intraganglionic ROS in DNA methylation of pro-nociceptive genes. We will examine whether ROS regulates DNA methylation of TRPV1 and TRPA1 genes in TG and whether stress elevates intraganglionic ROS, which maintains the reduced level of methylation of the pro-nociceptive genes. Our preliminary data suggest ROS as a key upstream factor involved in DNA methylation of the two pro-nociceptive genes. We predict that the blockade of ROS accumulation in TG or targeted methylation of DNA promoters will prevent stress-mediated potentiation of hyperalgesia and the upregulation of TRPV1 and TRPA1. Successful achievement of this project should unravel novel mechanisms involving DNA methylation and intraganglionic oxidative metabolites on functional regulation of multiple pro-nociceptive genes, providing a mechanistic basis for how inflammation and stress engage sensory ganglia to induce prolonged persistent muscle pain. The anticipated outcomes should have broad translational implications for the development of therapeutic approaches targeting transcriptional machineries that regulate DNA methylation of a specific pro-nociceptive gene or a cluster of genes sharing similar epigenetic mechanisms.

项目摘要 DNA甲基化是一种表观遗传因子，在调节基因表达和改变中起着重要作用。 DNA甲基化是与许多人类疾病相关的特征。炎症和环境诸如心理物理应激的因素诱导前伤害感受基因的去甲基化，导致它们的异常表情此更新应用程序的目的是远程调查肌肉炎症如何发生调节三叉神经节（TG）中多种前伤害感受基因的DNA甲基化，与疼痛和痛觉过敏有关我们的中心假设是咬肌炎症导致 TG中促伤害感受基因的甲基化降低，导致其异常表达，这有助于疼痛和机械性痛觉过敏的发展。我们进一步假设心理压力通过TG内过量产生活性氧（ROS）来增强这些作用， DNA甲基化在目标1中，我们将确定DNA甲基化在炎症疼痛反应中的作用。具体地说，我们将研究是否增加DNA甲基化通过DNMT在启动子区，个体的亲伤害感受基因，防止炎性疼痛和痛觉过敏。我们已确认促伤害感受基因如TRPV 1、TRPA 1、P2 X3和PIEZO 2含有结合DNMT的CG岛， DNMT活性的抑制增加了TG中DNMT的表达。为了确定DNA的作用在单个基因的甲基化中，我们设计并验证了一种新的DNMT融合蛋白复合物，特定基因的启动子区域，使用CRISPR-dCAS 9技术。我们期望， TG内的融合蛋白将阻止靶基因的上调，并揭示其相对贡献。在肌炎条件下疼痛和痛觉过敏的特定基因的DNA甲基化。在目标2中，我们将研究神经节内ROS在致伤害基因DNA甲基化中的作用。我们将研究 ROS是否调节TG中TRPV 1和TRPA 1基因的DNA甲基化以及应激是否升高神经节内ROS，其维持促伤害感受基因的甲基化水平降低。我们初步数据表明，ROS作为一个关键的上游因子，参与了两个前伤害感受基因的DNA甲基化，基因.我们预测，阻断TG中ROS的积累或DNA启动子的靶向甲基化，防止应激介导的痛觉过敏增强以及TRPV 1和TRPA 1的上调。成功该项目的成果应该解开涉及DNA甲基化和神经节内氧化代谢物对多种亲伤害感受基因的功能调节，提供了机制基础炎症和压力是如何影响感觉神经节，从而导致长时间的持续性肌肉疼痛。的预期的结果应该对治疗药物的开发具有广泛的转化意义，靶向调节特异性亲伤害感受因子的DNA甲基化的转录机制的方法一个基因或一组基因共享相似的表观遗传机制。