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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要虽然慢性疼痛代表了一个巨大的公共卫生问题，造成了惊人的经济损失仅在美国，驱动神经过度兴奋的分子机制每年就花费 560-6350 亿美元伤害感受途径的研究尚未完全明了。阐明初级感觉神经元的遗传异质性及其在神经后的可塑性然而，人们对这些神经元的全面分子特征知之甚少。尽管它们对疼痛很重要，但它们将伤害性信息从脊髓传递到大脑更好地理解脊髓投射神经元内基因表达的完整模式。可以揭示新的基于证据的策略来选择性抑制脊髓伤害性网络的输出作为缓解慢性疼痛的一种手段，长期目标是更好地了解神经和组织损伤的方式。改变中枢神经系统中伤害性回路的功能该应用的目的是识别损伤诱发的。脊髓投射神经元内基因表达的变化增强了慢性疼痛下的放电中心假设是上行脊髓投射神经元表现出独特的分子。受外周损伤显着调节以促进膜过度兴奋的表型。拟议工作的基本原理是鉴定在脊髓中优先表达的基因投射神经元将产生新的药理学方法来抑制伤害感受的上升流向大脑发送信息，最大限度地减少对大脑中整体感觉运动处理的不必要的干扰中心假设将通过追求以下具体目标来检验：（1）识别基因。成人脊髓内富含上行投射神经元；(2) 阐明在慢性疼痛条件下投射神经元的基因表达增加了膜的兴奋性。这些目标将通过使用翻译核糖体亲和纯化（TRAP）和下一代技术来实现 RNA测序技术与生物信息学、电生理学、免疫组织化学相结合和原位杂交方法是创新的，因为它将首次揭示，将脊髓伤害感受回路连接到小鼠大脑的那些神经元的遗传表型关键参与神经性疼痛和炎性疼痛的产生，并阐明如何该人群的分子特征在慢性疼痛状态下发生变化。研究将发现脊髓投射神经元的新的细胞类型特异性标记物以及识别潜在的遗传机制，通过这些机制外周损伤可以放大伤害感受的“增益” 因此，这项研究意义重大，因为它将揭示新的内容。分子靶标可被操纵以选择性地沉默上行脊髓投射神经元伤害，以引起安全有效的镇痛，同时最大限度地减少不良副作用。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Transcriptional profile of spinal dynorphin-lineage interneurons in the developing mouse.

发育中小鼠脊髓强啡肽谱系中间神经元的转录谱。

DOI：
10.1097/j.pain.0000000000001636
发表时间：
2019-06-03
期刊：
Pain
影响因子：
7.4
作者：
Elizabeth K. Serafin;Ale;er G Chamessian;er;Jie Li;Xiang Zhang;Am;a M. McGann;a;C. Brewer;T. Berta;M. Baccei
通讯作者：
M. Baccei

Single-nucleus characterization of adult mouse spinal dynorphin-lineage cells and identification of persistent transcriptional effects of neonatal hindpaw incision.

成年小鼠脊髓强啡肽谱系细胞的单核特征和新生儿后爪切口持续转录效应的鉴定。