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.

原发性纤毛是从几乎所有细胞表面伸出的短发状结构 脊椎动物的身体。在神经系统中，纤毛对发展和功能至关重要 神经元，包括脊髓，脑皮质，海马，黑质系统和 小脑。背根神经节（DRG）的感觉神经元详细阐述 纤毛出生在胚胎发生后不久，然后迁移到DRG中。 但是，关于主要纤毛在功能方面可能具有的潜在功能的知之甚少 成熟的DRG神经元。果蝇和大鼠的最新工作表现出刺猬的作用 伤害感受敏化中的信号传导。在哺乳动物中，刺猬途径证明了一个 依赖于原发性纤毛的信号转导。这项研究建议旨在测试 假设DRG的伤害性神经元表达的原发性纤毛调节伤害感受 从外围到达的信号，从而在正常下调节伤害感受信号传导 生理状况和病理生理状态。在特定目标1中，我们将 研究原发性纤毛在疼痛阈值以及炎症模型中的重要性 大鼠和小鼠模型的神经性疼痛。在特定目标2中，我们将测试 Hedghog信号传导在PIPN的阈值，炎症和慢性疼痛状态的调节中， 我们将检查刺猬信号传导对DRG神经元原发性纤毛的依赖性。 AIM 2检查了刺猬信号传导介导两者中的痛觉过敏的假设 炎症和PIPN疼痛状态。在大鼠中，靶向原代纤毛特异性flagellar的siRNA 运输（IFT）基因将通过鞘内注射给腰DRG。 siRNA- 这些基因的介导敲低特异性导致主要睫状功能的丧失。在 鼠标，表达集成到NAV1.8基因座的CRE重组酶的敲门线应 用作伤害性神经元中的IFT基因IFT88的工具。 两者中PIPN和刺猬信号的基于药理和siRNA的修饰 在 原发性纤毛和刺猬信号应采用。该项目的重要性是 我们将阐明刺猬作为慢性修饰的新信号转导途径 疼痛状态，以及其作用的基因座，是原发性纤毛。纤毛和 刺猬途径提供多种可药物的目标，可以证明可以转化为 用于疼痛疗法的新型治疗疗法。