Identification of novel analgesic targets in ascending spinal projection neurons

上行脊髓投射神经元中新型镇痛靶点的鉴定

• 批准号: 9486008
• 负责人: Mark L Baccei
• 金额: $ 23.99万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2020-05-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9486008
• 关键词: Absence of pain sensation; Accounting; Action Potentials; Adult; Adverse effects; Afferent Neurons; Affinity Chromatography; American; Analgesics; Automobile Driving; Axon; Bioinformatics; Brain; Cells; Chimeric Proteins; Clinical; Complement; Data; Development; Electrophysiology (science); Exhibits; G-Protein-Coupled Receptors; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Heterogeneity; Genetic Recombination; Goals; Human; Immunohistochemistry; In Situ Hybridization; In Vitro; Inflammation; Injury; Interneurons; Investigation; Ion Channel; Knowledge; Lesion; Mediating; Membrane; Messenger RNA; Modality; Molecular; Molecular Profiling; Molecular Target; Motor; Motor Neurons; Mus; Nerve; Nerve Tissue; Neurons; Nociception; Operative Surgical Procedures; Outcome; Output; Pain; Pain Research; Pathologic; Pathway interactions; Peripheral; Pharmacology; Phenotype; Physiological; Population; Posterior Horn Cells; Proprioception; Protein Kinase; Proteins; Public Health; Quality of life; Research; Research Personnel; Ribosomes; Role; Sensory; Spinal; Spinal Cord; Spinal cord posterior horn; Techniques; Testing; Time; Tissues; Touch sensation; Translating; Universities; Work; base; cell type; chronic pain; chronic painful condition; collaborative environment; cost; dorsal horn; economic cost; enhanced green fluorescent protein; evidence base; genetic profiling; in vivo; inflammatory pain; innovation; insight; molecular phenotype; nerve injury; neuronal excitability; next generation; novel; novel therapeutic intervention; pain perception; pain sensation; pain signal; painful neuropathy; patch clamp; response; sciatic nerve; sciatic nerve damage; sensory input; transcriptome sequencing; transmission process; Absence of pain sensation; Accounting; Action Potentials; Adult; Adverse effects; Afferent Neurons; Affinity Chromatography; American; Analgesics; Automobile Driving; Axon; Bioinformatics; Brain; Cells; Chimeric Proteins; Clinical; Complement; Data; Development; Electrophysiology (science); Exhibits; G-Protein-Coupled Receptors; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Heterogeneity; Genetic Recombination; Goals; Human; Immunohistochemistry; In Situ Hybridization; In Vitro; Inflammation; Injury; Interneurons; Investigation; Ion Channel; Knowledge; Lesion; Mediating; Membrane; Messenger RNA; Modality; Molecular; Molecular Profiling; Molecular Target; Motor; Motor Neurons; Mus; Nerve; Nerve Tissue; Neurons; Nociception; Operative Surgical Procedures; Outcome; Output; Pain; Pain Research; Pathologic; Pathway interactions; Peripheral; Pharmacology; Phenotype; Physiological; Population; Posterior Horn Cells; Proprioception; Protein Kinase; Proteins; Public Health; Quality of life; Research; Research Personnel; Ribosomes; Role; Sensory; Spinal; Spinal Cord; Spinal cord posterior horn; Techniques; Testing; Time; Tissues; Touch sensation; Translating; Universities; Work; base; cell type; chronic pain; chronic painful condition; collaborative environment; cost; dorsal horn; economic cost; enhanced green fluorescent protein; evidence base; genetic profiling; in vivo; inflammatory pain; innovation; insight; molecular phenotype; nerve injury; neuronal excitability; next generation; novel; novel therapeutic intervention; pain perception; pain sensation; pain signal; painful neuropathy; patch clamp; response; sciatic nerve; sciatic nerve damage; sensory input; transcriptome sequencing; transmission process

Project Summary/Abstract While chronic pain represents a massive public health problem with a staggering economic cost of $560-$635 billion each year in the U.S. alone, the molecular mechanisms driving neuronal hyperexcitability within nociceptive pathways remain incompletely understood. Significant progress has been made towards elucidating the genetic heterogeneity of primary sensory neurons and their plasticity in the aftermath of nerve or tissue damage. However, much less is known about the comprehensive molecular profile of those neurons that convey nociceptive information from the spinal cord to the brain, despite their clear importance for pain perception. A better understanding of the complete pattern of gene expression within spinal projection neurons could reveal new evidence-based strategies to selectively dampen the output of the spinal nociceptive network as a means to alleviate chronic pain. The long-term goal is to better understand how nerve and tissue damage alter the function of nociceptive circuits in the CNS. The objective of this application is to identify injury-evoked changes in gene expression within spinal projection neurons that enhance their firing under chronic pain conditions. The central hypothesis is that ascending spinal projection neurons exhibit a unique molecular phenotype that is significantly modulated by peripheral injury to promote membrane hyperexcitability. The rationale for the proposed work is that the identification of genes that are preferentially expressed in spinal projection neurons will yield new pharmacological approaches to suppress the ascending flow of nociceptive information to the brain, while minimizing unwanted disruptions to global sensorimotor processing within the spinal cord. The central hypothesis will be tested by pursuing the following specific aims: (1) Identify genes that are enriched in ascending projection neurons within the adult spinal cord; and (2) Elucidate changes in gene expression in projection neurons under chronic pain conditions that increase membrane excitability. These aims will be accomplished by using translating ribosome affinity purification (TRAP) and next generation RNA sequencing techniques in combination with bioinformatics, electrophysiological, immunohistochemical and in situ hybridization approaches. The proposed work is innovative because it will reveal, for the first time, the genetic phenotype of those neurons connecting the spinal nociceptive circuit to the mouse brain that are critically involved in the generation of neuropathic and inflammatory pain, as well as elucidate how the molecular signature of this population changes during the chronic pain state. The outcome of these investigations will be the discovery of new, cell type-specific markers of spinal projection neurons and the identification of potential genetic mechanisms by which peripheral injuries can amplify the “gain” of nociceptive transmission in the spinal cord. As a result, the proposed research is significant because it will reveal novel molecular targets which could be manipulated to selectively silence ascending spinal projection neurons after injury, in order to evoke safe and effective analgesia while minimizing undesirable side effects.

项目摘要/摘要 慢性疼痛代表了一个巨大的公共卫生问题，经济成本惊人 仅在美国，每年$ 560- $ 6350亿美元 在伤害性途径中仍然不完全理解。已经取得了重大进展 阐明主要感觉神经元的遗传异质性及其在神经后的可塑性 或组织损伤。但是，关于这些神经元的综合分子谱知之甚少 从脊髓到大脑传达伤害性信息 洞察力。更好地理解脊柱投射神经元中基因表达的完整模式 可以揭示新的基于证据的策略，以选择性地抑制脊柱伤害性网络的输出 作为减轻慢性疼痛的一种手段。长期目标是更好地了解神经和组织如何损害 改变中枢神经系统中伤害感受器的功能。该应用的目的是识别受伤诱发的 脊柱投射神经元中基因表达的变化，从而增强其在慢性疼痛下的发射 状况。中心假设是上升的脊柱投影神经元表现出独特的分子 外周损伤显着调节以促进膜过度刺激性的表型。这 拟议工作的理由是，鉴定在脊柱中最好表达的基因 投影神经元将产生新的药物方法来抑制伤害感受的上升流动 向大脑的信息，同时最大程度地减少对全球感觉运动处理的不良破坏 脊髓。中心假设将通过追求以下特定目的来检验：（1）识别基因 富含成年脊髓中升高投影神经元的人； （2）阐明变化 在慢性疼痛条件下投射神经元中的基因表达会增加膜的兴奋。 这些目标将通过使用翻译核糖体亲和力净化（TRAP）和下一代来实现 RNA测序技术与生物信息学，电生理，免疫组织化学结合 以及原位杂交方法。拟议的工作具有创新性，因为它将首次揭示 那些将脊柱伤害感受电路与小鼠脑连接的神经元的遗传表型 与神经性和炎症性疼痛的产生相关，并阐明 该人群的分子特征在慢性疼痛状态下变化。这些结果 调查将是发现脊柱投射神经元的新细胞类型特异性标记和 鉴定外围损伤可以扩大伤害感受的潜在遗传机制 在脊髓中的传播。结果，拟议的研究很重要，因为它将揭示新颖 可以操纵以选择性地沉默的分子靶标在脊柱投射神经元之后 受伤，以引起安全有效的镇痛，同时最大程度地减少不可避免的副作用。