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毛状结构。 脊椎动物的身体。在神经系统中，纤毛对于神经系统的发育和功能都至关重要 神经元，包括脊髓、大脑皮层、海马、黑质纹状体系统和 小脑。背根神经节 (DRG) 的感觉神经元详细阐述了单个初级 它们在胚胎发生过程中出生后不久，就会在体细胞中长出纤毛，并迁移到 DRG。 然而，对于初级纤毛在以下功能中可能具有的潜在功能一无所知 成熟的 DRG 神经元。最近对果蝇和大鼠的研究显示了 Hedgehog 的作用 伤害性敏化中的信号传导。在哺乳动物中，Hedgehog 通路表现出 信号转导依赖初级纤毛。本研究计划旨在测试 假设 DRG 伤害感受神经元表达的初级纤毛调节伤害感受 来自外围的信号，从而在正常情况下调节伤害性信号 生理条件和病理生理状态期间。在具体目标 1 中，我们将 研究初级纤毛在疼痛阈值和炎症模型中的重要性 大鼠和小鼠模型中的神经性疼痛。在具体目标 2 中，我们将测试以下角色： Hedghog 信号在 PIPN 阈值、炎症和慢性疼痛状态调节中的作用 我们将检查 Hedgehog 信号对 DRG 神经元初级纤毛的依赖性。 目标 2 检查 Hedgehog 信号传导介导两种疾病的痛觉过敏的假设 炎症和 PIPN 疼痛状态。在大鼠中，siRNA 靶向初级纤毛特异性鞭毛内 转运（IFT）基因将通过鞘内注射施用至腰椎DRG。 siRNA- 这些基因介导的敲低特别导致初级纤毛功能的丧失。在 小鼠，表达集成到 NaV1.8 位点的 Cre 重组酶的敲入系应 用作特异性消除伤害性神经元中的 IFT 基因 Ift88 的工具。 PIPN 和 Hedgehog 信号传导的药理学和基于 siRNA 的修饰 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 应采用初级纤毛和刺猬信号传导。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，用于修饰慢性 疼痛状态及其作用部位，即初级纤毛。纤毛和 Hedgehog 通路提供了多个可成药靶点，这些靶点可能被证明适合转化为 疼痛治疗的新疗法。