Profiling Translation in Nociceptor Plasticity

伤害感受器可塑性的分析翻译

• 批准号: 10046039
• 负责人: Zachary Campbell
• 金额: $ 40.35万
• 依托单位: UNIVERSITY OF TEXAS DALLAS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10046039
• 关键词: 5' Untranslated Regions; Acute; Address; Animals; Attenuated; Behavior; Chemicals; Chronic; Codon Nucleotides; Cues; Data; Detection; Development; Diabetic Neuralgia; Diabetic Neuropathies; Disease; Electrophysiology (science); Endoplasmic Reticulum; Event; Exposure to; FRAP1 gene; Generations; Genes; Genetic; Genome; Genomics; Goals; Healthcare Systems; High-Throughput Nucleotide Sequencing; Human; Hyperalgesia; Hypersensitivity; Immediate-Early Genes; Impairment; Inflammation Mediators; Inflammatory; Institute of Medicine (U.S.); Life; Link; Mechanics; Messenger RNA; Methods; Modeling; Molecular; Morals; Neuronal Plasticity; Neurons; Neuropathy; Neurosciences; Nociception; Nociceptors; Open Reading Frames; Pain; Pain management; Pathway interactions; Pattern; Peptides; Peripheral; Persistent pain; Pharmacology; Phase; Phenotype; Phosphorylation; Play; Population; Postoperative Pain; Production; Protein Biosynthesis; Pyruvaldehyde; RNA; Reading Frames; Regulation; Research; Resolution; Ribonucleases; Ribosomes; Role; Shapes; Signal Pathway; Signal Transduction; Spinal Ganglia; Structural Protein; Synaptic plasticity; Therapeutic Intervention; Tissues; Transcript; Translating; Translation Initiation; Translational Regulation; Translations; Treatment Efficacy; United States; Untranslated Regions; base; biological adaptation to stress; chronic pain; cytokine; density; endoplasmic reticulum stress; experimental study; genome-wide; genomic tools; improved; in vivo; inflammatory pain; inhibitor/antagonist; insight; interest; novel; painful neuropathy; programs; response; ribosome profiling; targeted treatment; transcriptome; translatome; virtual

Project Summary/Abstract Chronic pain is a pervasive and devastating condition. Translational control is a dominant theme in synaptic plasticity and plays a key role in pain plasticity, in particular the plasticity of peripheral nociceptors. Messenger RNA (mRNA) is subject to dynamic regulation by signaling pathways implicated in plasticity such as the integrated stress response (ISR) and mTOR. Yet, the identity of the mRNAs in nociceptors subjected to regulated translation during plasticity events is virtually unknown. Here, we use an exciting new genomics method termed ribosome profiling to gain genome-wide insight into translation control events in DRG neurons. Our preliminary studies reveal a rapid induction of translation of a specific subset of mRNAs that are known to be involved in neuronal plasticity in response to cytokine treatment of DRG neurons. Among them is the immediate early gene Arc. We identify S6K as required for activity dependent translation of Arc. A chemical inhibitor of S6K attenuates mechanical hypersensitivity in a model of inflammatory pain. Additionally, our preliminary data also indicate evidence for translation of a specific 5’ untranslated region (UTR) reading frame encoded by Calca/CGRP. The resulting peptide is able to promote pain amplification in vivo. In the first aim of this proposal, we will comprehensively characterize translation regulation in DRG neurons in response to cytokines with a focus on underlying mechanisms and targets of interest. We examine the function of a specific uORF with electrophysiology and pharmacology. In the second aim, we examine the effects of methylglyoxal (MGO) on translation. MGO is associated with diabetic neuropathy as well as discogenic neuropathies and our preliminary data indicate that its pain promoting effects depend on induction of the ISR. Interestingly, the ISR induces translation of uORFs suggesting this non-canonical form of translation as a new theme in neuropathic pain. We will probe the effects of MGO on translational control and determine if blocking the ISR is a viable option for neuropathic pain. Our experiments demonstrate the tremendous potential of a transformative genomics tool that enables new views on pain plasticity with astonishing molecular clarity.

项目概要/摘要 慢性疼痛是一种普遍且具有破坏性的疾病。翻译控制是突触的主导主题 可塑性，在疼痛可塑性，特别是外周伤害感受器的可塑性中起着关键作用。信使 RNA (mRNA) 受到与可塑性相关的信号通路的动态调节，例如 综合应激反应 (ISR) 和 mTOR。然而，伤害感受器中 mRNA 的身份受到调节 可塑性事件期间的翻译实际上是未知的。在这里，我们使用了一种令人兴奋的新基因组学方法，称为 核糖体分析，以获得对 DRG 神经元翻译控制事件的全基因组洞察。我们的初步 研究揭示了已知参与的特定 mRNA 子集的翻译的快速诱导 神经元可塑性响应 DRG 神经元的细胞因子治疗。其中有即早基因 弧。我们将 S6K 确定为 Arc 的活动相关翻译所需的。 S6K 的化学抑制剂会减弱 炎性疼痛模型中的机械超敏反应。此外，我们的初步数据还表明 由 Calca/CGRP 编码的特定 5' 非翻译区 (UTR) 阅读框翻译的证据。这 所得肽能够促进体内疼痛放大。在本提案的第一个目标中，我们将 全面表征 DRG 神经元响应细胞因子的翻译调节，重点关注 基本机制和利益目标。我们检查特定 uORF 的功能 电生理学和药理学。在第二个目标中，我们研究了甲基乙二醛（MGO）对 翻译。 MGO 与糖尿病性神经病以及椎间盘源性神经病有关，我们初步认为 数据表明，其疼痛促进作用取决于 ISR 的诱导。有趣的是，ISR 诱导 uORF 的翻译表明这种非规范的翻译形式是神经性疼痛的一个新主题。我们 将探讨 MGO 对翻译控制的影响，并确定阻断 ISR 是否是一个可行的选择 神经性疼痛。我们的实验证明了变革性基因组学工具的巨大潜力 以惊人的分子清晰度提供关于疼痛可塑性的新观点。