Profiling Translation in Nociceptor Plasticity

伤害感受器可塑性的分析翻译

基本信息

批准号：
10670777
负责人：
Zachary Campbell
金额：
$ 37.46万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
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/10670777
关键词：
5&apos Untranslated Regions Acute Address Animals Attenuated Behavior Binding Chemicals Chronic Codon Nucleotides Cues Data 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 Maps 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 Tissues Transcript Translating Translation Initiation Translational Regulation Translations United States Untranslated Regions biological adaptation to stress chronic pain chronic pain management cytokine detection method endoplasmic reticulum stress experimental study genome-wide genomic tools improved in vivo inflammatory pain inhibitor insight interest novel painful neuropathy pharmacologic programs response ribosome profiling targeted treatment therapeutically effective transcriptome translatome virtual

项目摘要

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。可塑性并在疼痛可塑性，特别是外周伤害感受器的可塑性中发挥关键作用。 RNA (mRNA) 受到与可塑性相关的信号通路的动态调节，例如然而，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“可塑性事件”。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。研究揭示了已知参与的特定 mRNA 子集的快速诱导翻译 DRG 神经元对细胞因子治疗的神经元可塑性就是其中之一。 Arc。我们确定 S6K 是 Arc 的活性依赖性翻译所必需的，S6K 的化学抑制剂会减弱。此外，我们的初步数据还表明，炎症性疼痛模型中存在机械过敏。由 Calca/CGRP 编码的特定 5' 非翻译区 (UTR) 阅读框翻译的证据。所得肽能够促进体内疼痛放大。在该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，重点关注我们通过以下方法检查特定 uORF 的功能。在第二个目标中，我们研究了甲基乙二醛（MGO）对电生理学和药理学的影响。 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。 uORF 的翻译表明这种非规范的翻译形式是神经性疼痛的新主题。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。