Identification of a novel DRG-specific long noncoding RNA and its role in neuropathic pain

新型 DRG 特异性长非编码 RNA 的鉴定及其在神经病理性疼痛中的作用

• 批准号: 10210612
• 负责人: Huijuan Hu
• 金额: $ 49.03万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10210612
• 关键词: Attenuated; Binding; Chronic; Clinical Trials; Code; Data; Development; Disease; Disease Management; Distress; Down-Regulation; Enzymes; Gene Expression; Genes; Genetic Transcription; Human; Hypersensitivity; Injury; Ion Channel; Laboratories; Lead; Length; Ligation; Maintenance; Mediating; Messenger RNA; Mus; Names; Neurons; Nucleotides; Opioid Receptor; POU Domain; Pain; Pain management; Peripheral nerve injury; Proteins; Public Health; RNA Polymerase II; Regulation; Role; Spinal Ganglia; Spinal nerve structure; Testing; Time; Transcription Coactivator; Transcription Factor 3; Untranslated RNA; Up-Regulation; Virus; Voltage-Gated Potassium Channel; chemokine; clinical application; cytokine; gene therapy; injured; knock-down; nerve injury; next generation; novel; novel strategies; overexpression; pain symptom; painful neuropathy; promoter; receptor; therapeutic target; transcription factor; transcriptome sequencing

Project Abstract Long noncoding RNAs (lncRNAs) regulate gene expression. Peripheral nerve injury dysregulated their expression in the pain-related regions including dorsal root ganglion (DRG). However, the role of most identified lncRNAs in neuropathic pain is still uncertain. Identifying novel lncRNAs and exploring their contribution to neuropathic pain may provide novel strategies for management of this disorder. We recently used a next generation RNA sequencing approach and identified a large, native, full-length non-coding RNA in mice and human. Because it is expressed highly in the DRG, we named it as DRG specific long noncoding RNA (DS-lncRNA). Our preliminary data revealed that peripheral nerve injury downregulated DS-lncRNA likely due to a decrease in the expression of a transcriptional activator Pou4f3 in the injured DRG. Rescuing this downregulation attenuated the nerve injury-induced pain hypersensitivity, likely through blockade of the increased interaction between RALY (a transcription co-factor) and the RNA polymerase II (RNA II) and consequent silence of the RALY/RNA II-triggered expression of Ehmt2 mRNA and its coding protein G9a (a key player in neuropathic pain) in the injured DRG. Given that DS-lncRNA can directly bind to RALY, our preliminary results indicate that DRG DS-lncRNA downregulation is required for neuropathic pain likely through negative regulation of DRG RALY/RNA II-triggered G9a expression. This proposal will further examine whether and how DS-lncRNA contributes to neuropathic pain. In Specific Aim 1, we will first investigate whether rescuing downregulation of DS-lncRNA in the injured DRG attenuates neuropathic pain development and maintenance. We will then examine whether mimicking nerve injury-induced downregulation of DRG DS-lncRNA leads to neuropathic pain symptoms in the absence of nerve injury. In Specific Aim 2, we will examine whether peripheral nerve injury results in time-dependent downregulation of DS-lncRNA and its transcription factor Pou4f3 in the DRG. We will also examine whether DS-lncRNA downregulation is attributed to a decrease of Pou4f3 expression in the injured DRG after peripheral nerve injury. In Specific Aim 3, we will test the effect of DS-lncRNA on the expression of Ehmt2 mRNA, G9a and their downstream pain-related genes in the injured DRG after peripheral nerve injury. We will also determine whether DS-lncRNA downregulation enhances the binding of RALY to RNA II leading to RALY/RNA II-triggered Ehmt2/G9a increase and G9a-controlled pain-related gene decrease in the injured DRG after peripheral nerve injury. Our study will likely identify a previously unknown regulatory mechanism for neuropathic pain. Given that virus- mediated gene therapy has been used in clinical trial, the present study will have a potential clinical application in neuropathic pain management.

项目摘要 长非编码 RNA (lncRNA) 调节基因表达。周围神经损伤使其失调 在包括背根神经节（DRG）在内的疼痛相关区域中表达。然而，大多数角色 已确定的神经性疼痛中的 lncRNA 仍不确定。识别新型 lncRNA 并探索它们 对神经性疼痛的贡献可能为治疗这种疾病提供新的策略。我们最近 使用下一代 RNA 测序方法并鉴定了一个大的、天然的、全长的非编码 RNA 老鼠和人类。由于它在DRG中高表达，我们将其命名为DRG特异性长非编码 RNA（DS-lncRNA）。我们的初步数据显示，周围神经损伤可能下调 DS-lncRNA 由于受损 DRG 中转录激活因子 Pou4f3 的表达减少。拯救这个 下调减弱了神经损伤引起的疼痛超敏反应，可能是通过阻断 RALY（转录辅助因子）和 RNA 聚合酶 II (RNA II) 之间的相互作用增强 随后 RALY/RNA II 的沉默触发了 Ehmt2 mRNA 及其编码蛋白 G9a（a 神经病理性疼痛的关键参与者）在受伤的 DRG 中。鉴于 DS-lncRNA 可以直接结合 RALY，我们的 初步结果表明，神经病理性疼痛可能需要 DRG DS-lncRNA 下调 通过 DRG RALY/RNA II 触发的 G9a 表达的负调节。该提案将进一步 检查 DS-lncRNA 是否以及如何导致神经性疼痛。在具体目标 1 中，我们首先 研究修复受损 DRG 中 DS-lncRNA 的下调是否可以减轻神经性疼痛 开发和维护。然后我们将检查是否模仿神经损伤引起的下调 在没有神经损伤的情况下，DRG DS-lncRNA 的 DRG DS-lncRNA 会导致神经性疼痛症状。在具体目标 2 中，我们 将检查周围神经损伤是否导致 DS-lncRNA 及其时间依赖性下调 DRG 中的转录因子 Pou4f3。我们还将检查 DS-lncRNA 下调是否归因于 周围神经损伤后受损 DRG 中 Pou4f3 表达减少。在具体目标 3 中，我们将 测试DS-lncRNA对Ehmt2 mRNA、G9a及其下游疼痛相关表达的影响 周围神经损伤后受损DRG中的基因。我们还将确定 DS-lncRNA 是否 下调增强 RALY 与 RNA II 的结合，导致 RALY/RNA II 触发 Ehmt2/G9a 周围神经损伤后受损DRG中G9a控制的疼痛相关基因增加，G9a控制的疼痛相关基因减少。我们的 研究可能会发现一种以前未知的神经性疼痛调节机制。鉴于该病毒—— 介导的基因治疗已用于临床试验，本研究将具有潜在的临床应用价值 在神经病理性疼痛治疗中的应用。