REST-mediated epigenomic and transcriptomic signatures in neuropathic pain

REST 介导的神经病理性疼痛的表观基因组和转录组特征

• 批准号: 10256781
• 负责人: SADHAN MAJUMDER
• 金额: $ 39.74万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10256781
• 关键词: ATAC-seq; Acute; Affect; Afferent Neurons; American; Animal Model; Attenuated; Behavioral Assay; Biochemical; Biological Markers; Biology; Cells; ChIP-seq; Chronic Phase; Clinical; Clinical Research; Collaborations; DNA; DNA-Directed RNA Polymerase; Data; Development; Disease; Down-Regulation; Drug Targeting; Epigenetic Process; Exhibits; G9a histone methyltransferase; Genes; Genetic; Genetic Transcription; Goals; Grant; Histones; Human; Hypersensitivity; Immunofluorescence Immunologic; Knockout Mice; Knowledge; Length; Malignant neoplasm of brain; Mediating; Messenger RNA; Methods; MicroRNAs; Molecular Probes; Mus; Nervous system structure; Neurons; Pain; Patients; Peripheral nerve injury; Potassium Channel; Predisposition; Prevention; Quality of life; Repression; Research; Resistance; Rest; Reverse Transcriptase Polymerase Chain Reaction; Role; Spinal Ganglia; System; Testing; Transcription Repressor; Traumatic Nerve Injury; United States National Institutes of Health; VP 16; Work; base; chemotherapy; chronic neuropathic pain; chronic pain; conditional knockout; epigenetic regulation; epigenetic silencing; epigenomics; experience; gender difference; genetic corepressor; genetic signature; genome-wide; genome-wide analysis; in vivo; inhibitor/antagonist; innovation; mouse model; mu opioid receptors; nerve injury; novel; overexpression; pain chronification; painful neuropathy; preclinical study; promoter; somatosensory; therapeutic target; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Chronic neuropathic pain is difficult to treat and remains a major clinical problem despite numerous clinical and preclinical studies. Our long-term goal is to identify new targets for treatment/prevention of chronic neuropathic pain. Mounting evidence suggests that epigenetic mechanisms and regulation via microRNAs (miRs) are associated with the transition from acute to chronic pain. The goal of this project is to identify REST- and G9a- related transcriptomic and epigenomic regulatory networks in the dorsal root ganglion (DRG) and their roles in the development of neuropathic pain. The transcriptional repressor REST is a major epigenomic regulator. In nerve injury-induced neuropathic pain, epigenetic silencing by REST overexpression in DRG neurons has been implicated in the repression of targets such as Oprm1, Kcnd3, Kcnq2, and scn10a genes. Histone methyl-transferase G9a is also an epigenomic corepressor of many transcription factors including REST. We recently found that G9a is critical in the epigenetic silencing of almost all K+ channel genes in DRG neurons via increased histone H3K9me2 on the genes' promoters during acute-to-chronic pain transition. Accordingly, mice with G9a ablated from DRG neurons do not develop chronic pain after nerve injury. Thus, both REST and G9a present suitable molecular probes to decipher the genetic and epigenetic bases of chronic pain. However, knowledge is limited about the transcriptomic and epigenomic networks associated with REST and G9a in neuropathic pain development. This multiple-PI grant is a collaboration between two labs with complementary expertise. In addition to the G9a mouse model described above, the two labs have developed an innovative experimental system consisting of complementary Rest conditional knock-out (cKO) and new conditional human REST overexpression (cOE) mouse models. Preliminary results indicate that whereas DRG-specific Rest cKO mice show attenuated pain hypersensitivity after nerve injury, DRG-specific REST cOE mice exhibit pain hypersensitivity. Thus, the two contrasting mouse models recapitulate the chronic pain transition, as anticipated based on work in the field, and provide an in vivo system to study transcriptional mechanisms of chronic pain. Here we propose to the test the central hypothesis that REST and G9a are involved in the development of chronic pain after DRG nerve injury via unique transcriptomic and epigenomic signatures. If successful, our proposed research will identify all REST and G9a targets in DRG neurons including mRNAs and miRs, as well as REST- and G9a-mediated epigenomic changes in the development of chronic pain after nerve injury. We will also functionally identify those REST- and G9a-regulated miRs that could be utilized to block REST- and G9a-mediated chronic pain. We will compare our results to those obtained in chemotherapy- induced chronic pain. In summary, the results from this work are likely to generate new signatures and biomarkers that could potentially be utilized to predict patient susceptibility or resistance to chronic pain development. The results could also be used to identify new actionable drug targets in chronic pain.

项目摘要 慢性神经性疼痛很难治疗，尽管许多临床和 临床前研究。我们的长期目标是确定治疗/预防慢性神经性的新目标 疼痛。越来越多的证据表明，通过microRNA（miR）的表观遗传机制和调节是 与从急性到慢性疼痛的过渡有关。该项目的目的是确定休息和G9A- 背根神经节（DRG）中的相关转录组和表观基因组调节网络及其在 神经性疼痛的发展。转录阻遏物休息是主要的表观基因组调节剂。在 神经损伤引起的神经性疼痛，DRG神经元中的静止过表达的表观遗传沉默具有 与OPRM1，KCND3，KCNQ2和SCN10A基因等靶标的抑制有关。组蛋白 甲基转移酶G9a也是许多转录因子（包括休息）的表观基因组核心。我们 最近发现，G9a在DRG神经元中几乎所有K+通道基因的表观遗传沉默至关重要 在急性到智力疼痛转变期间，基因启动子上的组蛋白H3K9ME2增加了。因此，老鼠 从DRG神经元中消融的G9A后，神经损伤后不会出现慢性疼痛。因此，休息和G9A都 存在合适的分子探针，以解读慢性疼痛的遗传和表观遗传底部。然而， 关于与REST和G9A相关的转录组和表观基因组网络的知识限制 神经性疼痛发育。这项多重拨款是两个实验室与互补的合作 专业知识。除了上述G9A鼠标模型外，两个实验室还开发了创新的 实验系统由互补的有条件淘汰（CKO）和新条件组成 人类休息过表达（COE）小鼠模型。初步结果表明，虽然DRG特定于 REST CKO小鼠显示神经损伤后的疼痛超敏反应，DRG特异性的REST COE小鼠展示 疼痛超敏反应。因此，两种对比的小鼠模型概括了慢性疼痛转变，如 根据该领域的工作进行预期，并提供了一个体内系统来研究的转录机制 慢性疼痛。在这里，我们建议测试中心假设，即休息和G9A参与 DRG神经损伤后通过独特的转录组和表观基因组特征的发育。如果 成功，我们拟议的研究将确定包括mRNA在内的DRG神经元中的所有休息和G9A靶标 和miR，以及慢性疼痛发育后的静止和G9A介导的表观基因组学变化 神经损伤。我们还将在功能上识别可以用来用于 阻塞静止和G9A介导的慢性疼痛。我们将将我们的结果与化学疗法中获得的结果进行比较 - 诱发慢性疼痛。总而言之，这项工作的结果可能会产生新的签名和 可能被用来预测患者敏感性或对慢性疼痛的抗性的生物标志物 发展。结果也可用于鉴定慢性疼痛中的新型可起作用的药物靶标。