Targeting Spinal Neuropeptide Y1 Receptor-expressing Neurons for the Control of Neuropathic Pain

靶向表达脊髓神经肽 Y1 受体的神经元来控制神经性疼痛

• 批准号: 10687806
• 负责人: Tyler Scott Nelson
• 金额: $ 4.78万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10687806
• 关键词: Ablation; Absence of pain sensation; Address; Affect; Afferent Neurons; Agonist; Americas; Amygdaloid structure; Analgesics; Behavior; Behavioral; Behavioral Symptoms; Brain; Brain region; Cells; Complex; Coupled; Data; Data Set; Development; Disease; Educational process of instructing; Electrophysiology (science); Equilibrium; Family; Fellowship; Fluorescent in Situ Hybridization; Frequencies; General Population; Genetic; Glutamates; Goals; Immediate-Early Genes; Injury; Institution; Interneurons; Knock-out; Label; Laboratories; Lead; Learning; Lesion; Maintenance; Measures; Mediating; Mentors; Mentorship; Messenger RNA; Methods; Mus; National Research Service Awards; Neurobiology; Neurons; Neuropathy; Neuropeptide Receptor; Neuropeptides; Neurosciences; Nociception; Nociceptors; Nucleosomes; Opioid; Output; Pain; Pathologic; Patients; Peripheral; Peripheral nerve injury; Pharmacology; Phase; Physiological; Population; Positioning Attribute; Principal Investigator; Proteins; Quality of life; Reporting; Research; Research Personnel; Signal Transduction; Slice; Societies; Spinal; Spinal Cord; Spinal cord posterior horn; Stimulus; System; Techniques; Testing; Therapeutic; Tissues; Training; Transgenic Organisms; United States; Universities; Work; Writing; behavior test; behavioral pharmacology; chronic pain; chronic painful condition; dorsal horn; experience; healing; improved; in vivo; in vivo calcium imaging; inflammatory pain; mouse model; nerve injury; neuropeptide Y; neuropeptide Y-Y1 receptor; novel strategies; optogenetics; pain perception; painful neuropathy; parabrachial nucleus; patch clamp; postsynaptic; receptor; relating to nervous system; sham surgery; single-cell RNA sequencing; skills; somatosensory; temporal measurement; tenure track; transcriptome sequencing; transcriptomics; voltage clamp; wireless

Project Summary Chronic pain conditions place significant burdens on patients, their families, and society by reducing quality of life and creating enormous financial consequences that total more than 630 billion USD annually for the United States of America alone. Neuropathic pain is a debilitating type of chronic pain that arises from a lesion or disease affecting the somatosensory system. Neuropathic pain affects 7-8% of the general population yet is poorly responsive to analgesic drugs, including opioids, thus, alternative therapeutics for treatment are desperately needed. However, the underlying mechanisms of the development and maintenance of neuropathic pain are poorly understood. It is hypothesized that neuropathic pain results from a loss of spinal cord dorsal horn inhibition and/or a gain in dorsal horn excitation that allows the propagation of low threshold innocuous inputs to be perceived as painful. Exactly how nerve injury disrupts this balance to generate a net pronociceptive tone, however, remains unclear. Specific Aim 1 describes promising preliminary data within our laboratory that implicates glutamatergic dorsal horn interneurons expressing the neuropeptide Y (NPY) Y1 receptor in both the development and maintenance of neuropathic pain. First, selective ablation of neuropeptide Y1 receptor- expressing interneurons (Y1-INs) with intrathecal NPY-saporin reduced the development of behavioral signs of neuropathic pain. Second, intrathecal pharmacology and intraspinal chemogenetic techniques indicate that Y1- INs are both necessary and sufficient for the behavioral manifestations of neuropathic pain. Lastly, both single cell RNA-sequencing and fluorescence in situ hybridization data indicate that Y1-INs segregate into three distinct dorsal horn interneuron subpopulations. Together, these observations form the premise for my central hypothesis that nerve injury increases the excitability of Y1-INs, and this makes one or more subpopulations of Y1-INs necessary for the behavioral symptoms of neuropathic pain. Specific Aim 2 will explore this hypothesis via intraspinal pharmacology, behavioral testing, in vivo wireless optogenetics, intersectional Cre-lox transgenics, and patch clamp electrophysiology. Together these methods will test which Y1-IN subpopulation(s) is/are necessary for the behavioral signs of neuropathic pain. Further, these methods will assess changes in pre- or postsynaptic excitatory and inhibitory activity to Y1-INs following nerve injury to uncover mechanistic changes in the circuit that might lead to the development of neuropathic pain. Specific Aim 3 details a plan to identify and pursue a neuroscience focused postdoctoral fellowship following the completion of the dissertation work described in Specific Aim 2. The overarching goals of this study are to increase our understanding of how nerve injury increases the excitability of Y1-IN subpopulations, and provide rationale for targeting spinal Y1-INs as a novel approach to treat neuropathic pain.

项目概要 慢性疼痛会降低患者的治疗质量，给患者及其家人和社会带来沉重负担。 生命并造成巨大的财务后果，每年给美国造成的损失总计超过 6300 亿美元 仅美利坚合众国。神经性疼痛是一种由病变或疾病引起的使人衰弱的慢性疼痛 影响体感系统。神经病理性疼痛影响着 7-8% 的普通人群，但症状较轻 对镇痛药物（包括阿片类药物）有反应，因此迫切需要替代疗法 需要。然而，神经性疼痛发生和维持的根本机制是 不太了解。据推测，神经性疼痛是由于脊髓背角抑制丧失所致 和/或背角激励增益，允许低阈值无害输入的传播 被认为是痛苦的。神经损伤到底是如何破坏这种平衡从而产生净伤害性音调的， 然而，目前尚不清楚。具体目标 1 描述了我们实验室内有希望的初步数据， 暗示谷氨酸能背角中间神经元在两个细胞中表达神经肽Y（NPY）Y1受体 神经性疼痛的发展和维持。一、选择性消融神经肽Y1受体—— 鞘内注射 NPY-皂草素可减少表达中间神经元 (Y1-IN) 的行为体征的发展 神经性疼痛。其次，鞘内药理学和椎管内化学遗传学技术表明，Y1- IN 对于神经性疼痛的行为表现来说既是必要的也是充分的。最后，两人都是单身 细胞 RNA 测序和荧光原位杂交数据表明 Y1-IN 分为三种不同的 背角中间神经元亚群。这些观察结果共同构成了我的中心思想的前提 假设神经损伤增加了 Y1-IN 的兴奋性，这使得一个或多个亚群 Y1-IN 是神经性疼痛行为症状所必需的。具体目标 2 将探讨这一假设 通过椎管内药理学、行为测试、体内无线光遗传学、交叉 Cre-lox 转基因学和膜片钳电生理学。这些方法将一起测试哪些 Y1-IN 亚群 对于神经性疼痛的行为体征来说是必要的。此外，这些方法将评估预 或神经损伤后 Y1-IN 的突触后兴奋和抑制活性，以揭示机制变化 在可能导致神经性疼痛发生的回路中。具体目标 3 详细说明了确定的计划 并在完成论文工作后寻求以神经科学为重点的博士后奖学金 具体目标 2 中描述。这项研究的总体目标是加深我们对神经如何 损伤增加了 Y1-IN 亚群的兴奋性，并为将脊髓 Y1-IN 作为靶向药物提供了理论基础。 治疗神经性疼痛的新方法。