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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：
9895424
负责人：
JIN Y Ro
金额：
$ 36.69万
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 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9895424
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.

项目总结