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Regulation of Nerve Injury-induced Gene Expression in Neuropathic Pain

神经病理性疼痛中神经损伤诱导的基因表达的调节

基本信息

批准号：
10188652
负责人：
Zhonghui Guan
金额：
$ 34.67万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10188652
关键词：
Adult Afferent Neurons Amplifiers Animals Antibodies B-Cell Lymphomas Binding Biochemical Process CSF1 gene CSF1R gene Cells Data Development Diabetic Neuropathies Economic Burden Embryo Embryonic Development Epigenetic Process Female Gene Expression Genes Genetic Transcription Histones Hour Hypersensitivity Immunohistochemistry Impairment Injury Intrathecal Injections Knock-out Local Anesthetics Macrophage Colony-Stimulating Factor Malignant Neoplasms Mechanics Microglia Molecular Target Motor Neurons Mus Neurons Operative Surgical Procedures Perioperative Persistent pain Phenotype Postoperative Pain Preemptive Analgesia Process Quality of life Quantitative Reverse Transcriptase PCR Regulation Reverse Transcriptase Polymerase Chain Reaction Sensory Ganglia Series Signal Transduction Spinal Cord Spinal Ganglia Surgical Injuries Techniques Testing Time Tissues Trauma Traumatic Nerve Injury Up-Regulation base c-myc Genes chromatin immunoprecipitation cytokine epigenetic marker epigenetic regulation experimental study gene induction genome browser inhibitor/antagonist injured macrophage male mouse model nerve injury neutralizing antibody pain behavior painful neuropathy post-trauma preemptive intervention prevent promoter recruit sciatic nerve transcription factor

项目摘要

Abstract Nerve injury-induced neuropathic pain following surgery or trauma has a significant economic burden that impairs quality of life. Despite the identification of a host of molecular targets implicated in the development of neuropathic pain, post-surgical and post-trauma neuropathic pains are often intractable. Unlike other neuropathic pain conditions, such as diabetic neuropathy, the time that nerve injury occurs during surgery or following trauma is well defined. As a result, it is possible that a pre-emptive intervention, either during the perioperative period or shortly after the trauma, could prevent the development of persistent pain. Indeed, local anesthetic based pre-emptive analgesia techniques have been developed. However, although these techniques are very good in managing immediate post-surgical pain, they only offer limited long-term benefit. In a mouse model of neuropathic pain in which the sciatic nerve is surgically injured, we recently demonstrated a critical contribution of injury-induced de novo synthesis and release of the cytokine, colony stimulating factor 1 (CSF1, also known as macrophage colony stimulating factor) from dorsal root ganglion (DRG) sensory neurons. Our studies established that CSF1 is both necessary and sufficient for nerve injury-induced activation of spinal cord microglia, the tissue macrophages of the CNS, and for post-injury mechanical hypersensitivity/ neuropathic pain behavior. Here, in Specific Aims 1 and 2, we propose experiments that will first define the transcriptional mechanism that triggers CSF1 induction in injured DRG neurons. Our preliminary analysis of the Santa Cruz Genome Browser ChIP-Seq data from B-cell lymphoma revealed 19 transcription factors that bind to CSF1 promoter. Among these 19 genes, using quantitative RT-PCR we determined that c-Myc is also induced in DRG after nerve injury. Interestingly, the c-Myc induction occurred within hours of nerve injury, well before the induction of CSF1. Immunohistochemistry confirmed the induction of c-Myc and its co-expression with CSF1 in DRG neurons. Of note, c-Myc is a universal amplifier of gene expression in cancer and embryonic development, but its function in adult non-proliferating cells is largely unknown. More importantly, pre-emptive local anesthetic treatment of sciatic nerve prior to the injury did not prevent the induction of c-Myc in DRG neurons, indicating that its induction is not activity dependent. Based on these findings, we hypothesize that c-Myc is a major contributor to CSF1 gene induction in the injured DRG neurons, and that targeting c-Myc might prevent the CSF1 gene induction and thus prevent neuropathic pain development. Finally, as the expression of CSF1 persists for weeks after nerve injury, we will also test the hypothesis that CSF1 not only contributes to the initiation of the neuropathic pain phenotype, but also to its persistence. Thus in Specific Aim 3, we will delete or block CSF1 from DRG neurons after injury occurs. Finally, given recent evidence for differences in microglial contribution to the neuropathic pain phenotype in male and female mice, our studies will also assess the contribution of these mechanisms both in male and female mice.

摘要手术或创伤后神经损伤引起的神经性疼痛具有显著的经济负担，影响生活质量。尽管鉴定出了许多与肿瘤发生有关的分子靶点，神经性疼痛、手术后和创伤后神经性疼痛通常是难治的。不像其他神经性疼痛病症，如糖尿病性神经病变，在手术期间发生神经损伤的时间，或创伤后的症状是明确的。因此，无论是在战争期间，围手术期或创伤后不久，可以防止持续性疼痛的发展。事实上，当地已经开发了基于麻醉剂的超前镇痛技术。然而，尽管这些技术在治疗术后立即疼痛方面非常好，但它们只能提供有限的长期益处。在一个神经病理性疼痛的小鼠模型中，坐骨神经被手术损伤，我们最近证明，损伤诱导的细胞因子集落刺激因子的从头合成和释放的关键贡献 1（CSF 1，也称为巨噬细胞集落刺激因子），来自背根神经节（DRG）感觉神经元我们的研究表明，CSF1是神经损伤诱导激活的必要和充分条件脊髓小胶质细胞，CNS的组织巨噬细胞，以及损伤后机械超敏反应/ 神经性疼痛行为在这里，在具体目标1和2中，我们提出了实验，首先将定义在受损的DRG神经元中触发CSF 1诱导的转录机制。我们初步分析了圣克鲁斯基因组浏览器B细胞淋巴瘤的ChIP-Seq数据揭示了19种结合的转录因子 CSF1启动子。在这19个基因中，使用定量RT-PCR，我们确定c-Myc也是在神经损伤后的DRG中诱导。有趣的是，c-Myc的诱导发生在神经损伤后的几个小时内，在诱导CSF 1之前。免疫组化证实了c-Myc的诱导及其共表达与CSF 1在DRG神经元中的表达。值得注意的是，c-Myc是癌症中基因表达的通用放大器，胚胎发育，但其在成人非增殖细胞中的功能在很大程度上是未知的。更重要的是，在损伤前对坐骨神经进行预先局部麻醉治疗并不能阻止c-Myc的诱导在DRG神经元中，表明其诱导不是活动依赖性的。基于这些发现，我们假设c-Myc是损伤的DRG神经元中CSF 1基因诱导的主要贡献者，靶向c-Myc可能阻止CSF1基因诱导，从而阻止神经性疼痛的发展。最后，由于CSF 1的表达在神经损伤后持续数周，我们还将检验以下假设： CSF 1不仅有助于神经性疼痛表型的起始，而且有助于其持续性。因此在具体目标3中，我们将在损伤发生后从DRG神经元中删除或阻断CSF 1。最后，鉴于最近在雄性和雌性小鼠中小胶质细胞对神经性疼痛表型的贡献的差异的证据，我们的研究还将评估这些机制在雄性和雌性小鼠中的作用。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Modified Spared Nerve Injury Surgery Model of Neuropathic Pain in Mice.

小鼠神经性疼痛的改良保留神经损伤手术模型。

DOI：
10.3791/63362
发表时间：
2022
期刊：
Journal of visualized experiments : JoVE
影响因子：
0
作者：
He,Liangliang;Zhao,Wenxing;Zhang,Lingyi;Ilango,Maalveka;Zhao,Na;Yang,Liqiang;Guan,Zhonghui
通讯作者：
Guan,Zhonghui

Ion Channels in Anesthesia.

麻醉中的离子通道。