3’ End Regulation in Nociceptor Plasticity

项目总结/摘要 管理不善的疼痛对我们的医疗保健系统造成了巨大的负担，并产生了巨大的人类健康风险。 痛苦受伤后，疼痛是由炎性细胞因子的产生引起的，炎性细胞因子诱导炎症反应的变化。 伤害感受器神经元的兴奋性。更好地理解促进伤害感受器的分子机制 可塑性对于改善疼痛治疗至关重要。转录后基因调控已成为一个主导主题 疼痛诱发的可塑性聚腺苷酸结合蛋白（Poly（A）binding proteins，PABP）是一个保守的3'端结合蛋白家族， 调节翻译起始并在发育和记忆中起重要作用的因子。我们将 使用无偏的下一个- 世代排序法这些信息将用于产生一类新的化学稳定的 RNA称为特异性衍生的竞争性抑制剂寡核苷酸（SPOT-ON）。与基因组编辑或 RNA干扰，SPOT-ON耐受性良好，并绕过对宿主因子的需要，因为它们的功能是 “诱饵”与靶向基因破坏相反，药理学抑制的主要优点是能够 在一个更大的基因家族中同时损害多个同源物。我们的初步调查结果表明，一个双- 特异性PABP SPOT-ON在体内提供了强有力的抗痛觉过敏作用。基于这一发现，我们假设 PABP是伤害感受器可塑性的重要介质。本文介绍了SPOT的初步发展， 减少伤害感受器中的蛋白质合成并特异性作用于poly（A）介导的mRNA翻译的ON (Aim 1）。我们通过检测PABP的局部翻译和全局翻译来探讨其作用机制。 识别伤害感受器中的靶点（目标2）。最后，我们确定PABP抑制如何影响伤害感受器 对损伤的兴奋性和行为反应（目标3）。

Project Summary/Abstract Poorly managed pain creates an enormous burden on our healthcare system and produces tremendous human suffering. Following an injury, pain is caused by the production of inflammatory cytokines that induce changes in the excitability of nociceptor neurons. A better understanding of molecular mechanisms that facilitate nociceptor plasticity is vital for improved pain treatment. Post-transcriptional gene control has emerged as a dominant theme in pain induced plasticity. We focus on Poly(A) binding proteins (PABPs), a conserved family of 3’ end associated factors that regulate translation initiation and play prominent roles in development and memory. We will determine binding specificities for PABPs present in dorsal root ganglion (DRG) neurons using an unbiased next- generation sequencing approach. This information will be used to generate a novel class of chemically stabilized RNAs called specificity-derived competitive inhibitor oligonucleotides (SPOT-ONs). Unlike genome editing or RNA interference, SPOT-ONs are well tolerated and bypass the need for host-factors as they function as “decoys”. A major advantage of pharmacological inhibition as opposed to targeted gene disruption is the ability to simultaneously impair multiple homologues in a larger gene family. Our preliminary findings indicate that a bi- specific PABP SPOT-ON provides robust anti-hyperalgesic effects in vivo. Based on this finding, we hypothesize that PABPs are crucial mediators of plasticity in nociceptors. We describe preliminary development of SPOT- ONs which reduce protein synthesis in nociceptors and act specifically on poly(A)-mediated mRNA translation (Aim 1). We probe PABP mechanism of action through examination of localized translation and global identification of targets in nociceptors (Aim 2). Finally, we determine how PABP inhibition impacts nociceptor excitability and behavioral responses to injury (Aim 3).