Role of Nociceptor Primary Cilia in Inflammatory and Neuropathic Pain

伤害感受器初级纤毛在炎症和神经性疼痛中的作用

• 批准号: 10575524
• 负责人: KERRY L TUCKER
• 金额: $ 39.05万
• 依托单位: UNIVERSITY OF NEW ENGLAND
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-23 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10575524
• 关键词: Acute; Address; Adult; Afferent Neurons; Animals; Axon; Binding; Brain; Carrier Proteins; Cell surface; Cells; Cerebellum; Cerebral cortex; Cilia; Complex; Corpus striatum structure; Cre driver; Data; Dependence; Development; Drosophila genus; Electrophysiology (science); Embryonic Development; Enterobacteria phage P1 Cre recombinase; Erinaceidae; Family; Genes; Genetic Transcription; Hair; Hippocampal Formation; Hippocampus (Brain); Hyperalgesia; Inflammatory; Intrathecal Injections; Knock-in; Ligands; Mammals; Mediating; Modeling; Modification; Mus; Mutant Strains Mice; Nervous system structure; Neurons; Nociception; Nociceptors; Organelles; Pain; Pain Threshold; Pain management; Pathway interactions; Pharmacology; Physiological; Play; Process; Proteins; Rattus; Research; Research Proposals; Rodent Model; Role; Signal Transduction; Signal Transduction Pathway; Small Interfering RNA; Spinal Cord; Spinal Ganglia; Structure; Substantia nigra structure; Surface; System; Testing; Translations; Work; anterograde transport; base; body system; chronic pain; conditional knockout; druggable target; gain of function; hedgehog signal transduction; inflammatory pain; information processing; innovation; knock-down; loss of function; mouse model; mutant; neuronal cell body; nigrostriatal system; novel therapeutics; pain signal; painful neuropathy; receptor; response; retrograde transport; smoothened signaling pathway; tool

Primary cilia are short hair-like structures protruding from the surface of almost all cells in the vertebrate body. In the nervous system, cilia are critical for both the development and the function of neurons, including in spinal cord, cerebral cortex, hippocampus, the nigrostriatal system, and the cerebellum. Sensory neurons of the dorsal root ganglia (DRG) elaborate a single primary cilium at their soma shortly after they are born in embryogenesis and migrate into the DRG. However, nothing is known about potential functions that primary cilia may have in the function of mature DRG neurons. Recent work in the fruit fly and in the rat have shown a role for Hedgehog signaling in nociceptive sensitization. In mammals, the Hedgehog pathway demonstrates a dependence upon primary cilia for signal transduction. This research proposal seeks to test the hypothesis that primary cilia expressed by nociceptive neurons of the DRG modulate nociceptive signals arriving from the periphery and thereby modulate nociceptive signaling under normal physiological conditions and during pathophysiological states. In Specific Aim 1 we shall investigate the importance of primary cilia in pain thresholds and in models of inflammatory and neuropathic pain in both rat and mouse models. In Specific Aim 2 we shall test the role of Hedghog signalling in the modulation of threshold, inflammatory, and chronic pain states in PIPN, and we shall examine the dependence of Hedgehog signalling upon primary cilia of DRG neurons. Aim 2 examines the hypothesis that Hedgehog signaling mediates hyperalgesia in both inflammatory and PIPN pain states. In the rat, siRNA targeting primary cilium-specific intraflagellar transport (IFT) genes will be administered to lumbar DRGs through intrathecal injection. siRNA- mediated knockdown of these genes specifically leads to a loss of the primary ciliary function. In the mouse, knock-in lines expressing a Cre recombinase integrated into the NaV1.8 locus shall be used as a tool to specifically eliminate the IFT gene Ift88 in nociceptive neurons. Pharmacological and siRNA-based modification of PIPN and Hedgehog signaling in both knockdown and wildtype rats, as well as loss-of-function and gain-of-function mouse mutants in primary cilia and in Hedgehog signaling, shall be employed. The significance of this project is that we shall elucidate Hedgehog as a new signal transduction pathway for the modification of chronic pain states, and also the locus at which it acts, the primary cilium. Both the cilium and the Hedgehog pathway offer multiple druggable targets that may prove amenable for translation into novel therapeutics for pain therapy.

初级纤毛是从几乎所有细胞表面突出的短发状结构， 脊椎动物的身体在神经系统中，纤毛对发育和功能都至关重要 神经元，包括脊髓，大脑皮层，海马，黑质纹状体系统， 小脑背根神经节（DRG）的感觉神经元阐述了一个单一的初级 纤毛在它们的索马体中在胚胎发生中出生后不久就迁移到DRG中。 然而，对于初级纤毛在细胞功能中可能具有的潜在功能， 成熟DRG神经元。最近在果蝇和大鼠中的研究表明，刺猬蛋白在 在伤害性敏化中的信号传导。在哺乳动物中，Hedgehog通路显示了一种 依赖初级纤毛进行信号转导。这项研究旨在测试 背根神经节伤害感受神经元表达的初级纤毛调节伤害感受的假设 来自外周的信号，从而在正常情况下调节伤害感受信号 生理条件和病理生理状态期间。在具体目标1中， 研究初级纤毛在疼痛阈值和炎症模型中的重要性， 在大鼠和小鼠模型中的神经性疼痛。在具体目标2中，我们将测试 Hedghog信号在PIPN中调节阈值、炎症和慢性疼痛状态， 我们将研究Hedgehog信号对DRG神经元初级纤毛的依赖性。 目的2检验Hedgehog信号通路介导两种神经元痛觉过敏的假说。 炎症和PIPN疼痛状态。在大鼠中，靶向初级纤毛特异性鞭毛内的siRNA 将通过鞘内注射向腰椎DRG施用IFT基因。siRNA- 介导的这些基因的敲除特异性地导致初级纤毛功能的丧失。在 表达整合到NaV1.8基因座中的Cre重组酶的小鼠敲入系应 可用作特异性消除伤害感受神经元中IFT基因Ift 88的工具。 PIPN和Hedgehog信号传导的药理学和基于siRNA的修饰在两种疾病中的应用 敲低和野生型大鼠，以及功能丧失和功能获得的小鼠突变体， 初级纤毛和Hedgehog信号传导中的纤毛。这个项目的意义在于， 我们将阐明Hedgehog作为一种新的信号转导途径，用于慢性炎症的调节， 疼痛状态，以及它作用的部位，初级纤毛。纤毛和 Hedgehog途径提供了多个可药物化的靶点，这些靶点可能被证明适合于翻译成 用于疼痛治疗的新疗法。