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.

项目摘要 慢性疼痛状况通过降低质量的质量，给患者，家人和社会带来巨大负担 生活和产生巨大的财务后果，曼联每年总计超过6,300亿美元 仅美国国家。神经性疼痛是一种令人衰弱的慢性疼痛类型，是由病变或疾病引起的 影响体感系统。神经性疼痛影响7-8％的普通人群，但很差 对包括阿片类药物在内的镇痛药的反应，因此，治疗的替代治疗方法是拼命的 需要。但是，神经性疼痛的发展和维持的基本机制是 理解不佳。假设神经性疼痛是由于脊髓背角抑制的丧失而引起的 和/或背喇叭激发的增益，允许低阈值无害输入的传播 被认为很痛苦。确切的神经损伤如何破坏这种平衡以产生净引起感人的音调， 但是，尚不清楚。特定目标1描述了我们实验室中有希望的初步数据 暗示在两个中表达​​神经肽Y（NPY）Y1受体的谷氨酸能背角中神经元 神经性疼痛的发展和维持。首先，神经肽Y1受体的选择性消融 用鞘内NPY-糖果表达中间神经元（Y1-INS）减少了行为迹象的发展 神经性疼痛。其次，鞘内药理学和脊髓内化学发生技术表明Y1- INS对于神经性疼痛的行为表现既需要又足够。最后，都单身 细胞RNA的测序和荧光原位杂交数据表明，y1-Ins分为三种不同 背喇叭中间神经元亚群。这些观察结果在一起是我中央的前提 假设神经损伤会增加Y1-INS的兴奋性，这使得 Y1-INS对于神经性疼痛的行为症状所必需的。具体目标2将探讨这一假设 通过脊柱内药理学，行为测试，体内无线光遗传学，交叉Cre-lox 转基因和斑块夹电生理学。这些方法共同测试哪种Y1-IN亚群（S） 对于神经性疼痛的行为迹象是必要的。此外，这些方法将评估预审前的变化 神经损伤后Y1-IN的突触后兴奋性和抑制活性，以发现机械变化 在可能导致神经性疼痛发展的电路中。特定目标3详细识别计划 并在完成论文工作后追求以神经科学为重点的博士后研究金 在特定目标2中描述的。这项研究的总体目标是提高我们对神经的理解 损伤增加了Y1-IN亚群的兴奋性，并为将脊柱Y1 INS作为A提供了理由 治疗神经性疼痛的新方法。