Amygdala pain mechanisms

杏仁核疼痛机制

基本信息

批准号：
10390809
负责人：
Volker Neugebauer
金额：
$ 55.16万
依托单位：
TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-15 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10390809
关键词：
Acute Acute Pain Address Affective Amygdaloid structure Anxiety Area Astrocytes Basic Science Behavior Behavioral Behavioral Assay Binding Bioinformatics Brain Cell Nucleus Cells Chronic Clinical Complex Corticotropin-Releasing Hormone Development Disease Electrophysiology (science)Elements Emotional Emotions Female Funding Genes Healthcare Immunohistochemistry Individual Interdisciplinary Study Knowledge Label Lateral Ligation Measures Membrane Mental Depression Microglia Modeling Molecular Molecular Biology Molecular Profiling Molecular Target Neuraxis Neuroglia Neuroimmune Neuronal Plasticity Neurons Nociception Outcome Measure Output Pain Pain management Pathway interactions Persistent pain Pharmaceutical Preparations Pharmacology Phase Physiology Play Pontine structure Property Rattus Regulation Research Project Grants Resolution Role Sensory Sensory Thresholds Signal Transduction Slice Spinal Spinal nerve structure Spine pain Sucrose Synapses Synaptic Transmission System Testing Therapeutic Time Transgenic Organisms United States National Institutes of Health Viral Viral Vector Work base cell type chronic neuropathic pain chronic pain forced swim test improved innovation insight interdisciplinary approach male molecular drug target molecular marker novel optogenetics pain behavior pain chronification pain model pain relief painful neuropathy patch clamp peripheral pain preference protein expression public health relevance response sexual dimorphism single-cell RNA sequencing spontaneous pain tool transcriptomics vocalization

项目摘要

Project Summary Chronic pain, a complex multidimensional disorder, remains a major health care issue and a therapeutic challenge. The development of new and improved therapeutic strategies requires the full understanding of mechanisms of chronic pain at all levels of the neuraxis and for all cell types, including non-neuronal elements. Neuroimmune signaling has emerged as a peripheral and spinal pain mechanism, but little is known about the role, regulation and therapeutic potential of molecular crosstalk between neuronal and glial cell types in the brain in the context of pain. To address this important knowledge gap we will build on our NIH-funded work (since 1999) that impacted the field by identifying neuroplasticity in the amygdala, a brain center for emotions, as a critical mechanism for emotional-affective aspects of pain and pain modulation. The proposed project will test the novel hypothesis that chronification of amygdala plasticity and neuropathic pain behaviors depends on a cascade of neuroimmune signaling that can be targeted to mitigate neuropathic pain. Specifically, enhanced synaptic drive of certain amygdala neuron types at the acute pain stage activates different types of glia, and glia-derived factors generate hyperexcitability in distinct amygdala neuron types at the chronic stage to maintain neuropathic pain. A comprehensive multidisciplinary approach will be used that integrates state-of-the-art transcriptomics, bioinformatics, behavioral assays, brain slice electrophysiology, pharmacology, chemogenetics, optogenetics, viral vector strategies, immunohistochemistry and molecular biology for the analysis of neuroimmune interactions within and between different types of neuronal and glial cells in the amygdala output region (central nucleus, CeA) in the well-established spinal nerve ligation (SNL) rat model of neuropathic pain. Male and female rats will be studied. A transgenic Crh-Cre rat model will be used to study the CeA corticotropin releasing factor (CRF) system, an important player in amygdala plasticity, and its role in neuroimmune signaling. Aim 1 will use a combination of cell-specific bulk and single-cell RNA sequencing (scRNA-Seq) to identify individual genes, molecular pathways and functional cellular states associated with pain chronification. A novel computational scRNA-Seq- based interactome analysis will determine neuron-glia interactions at the acute and chronic stages of neuropathic pain and identify drug-targetable molecular factors. Aim 2 will determine the behavioral significance of neuron-glia-neuron signaling in the CeA at different stages of neuropathic pain, using chemogenetic activation and inhibition of different cell types and pharmacological (or viral based) tools to modulate molecular factors. Sensory thresholds, emotional responses, non-evoked ongoing pain, and anxiety- and depression-like behaviors will be measured. Aim 3 will determine electrophysiological mechanisms of neuron-glia-neuron signaling in the CeA at different stages of neuropathic pain, using patch-clamp recordings of different cell types in brain slices and chemogenetic and pharmacological (or viral based) manipulations. Successful completion of these conceptually innovative studies will significantly advance our knowledge of neuro-immune signaling in brain plasticity and in the transition from acute to chronic pain and provide novel targets to mitigate chronic neuropathic pain.

项目概要慢性疼痛是一种复杂的多维疾病，仍然是一个主要的卫生保健问题和治疗挑战。这开发新的和改进的治疗策略需要充分了解慢性疼痛的机制在神经轴的各个层面和所有细胞类型，包括非神经元元件。神经免疫信号已经出现作为一种外周和脊髓疼痛机制，但人们对它的作用、调节和治疗潜力知之甚少。在疼痛的情况下，大脑中神经元和神经胶质细胞类型之间的分子串扰。为了解决这个重要的我们将在 NIH 资助的工作（自 1999 年以来）的基础上弥补知识差距，该工作通过识别神经可塑性影响了该领域在杏仁核中，大脑的情绪中心，作为疼痛和疼痛的情绪情感方面的关键机制调制。拟议的项目将测试新的假设，即杏仁核可塑性的时间化和神经性疼痛行为取决于一系列神经免疫信号传导，可以有针对性地缓解神经性疼痛。具体来说，在急性疼痛阶段某些杏仁核神经元类型的突触驱动增强激活不同类型的神经胶质细胞，神经胶质细胞衍生因子在不同的杏仁核神经元类型中产生过度兴奋性慢性阶段维持神经性疼痛。将采用全面的多学科方法整合了最先进的转录组学、生物信息学、行为分析、脑切片电生理学、药理学、化学遗传学、光遗传学、病毒载体策略、免疫组织化学和分子生物学分析杏仁核中不同类型神经元和神经胶质细胞内部和之间的神经免疫相互作用在成熟的脊髓神经结扎（SNL）大鼠神经性疼痛模型中，输出区域（中央核，CeA）。将研究雄性和雌性大鼠。转基因 Crh-Cre 大鼠模型将用于研究 CeA 促肾上腺皮质激素释放因子（CRF）系统，杏仁核可塑性的重要参与者，及其在神经免疫信号传导中的作用。目标1 将结合使用细胞特异性批量测序和单细胞 RNA 测序 (scRNA-Seq) 来识别单个基因，与疼痛慢性相关的分子途径和功能细胞状态。一种新颖的计算 scRNA-Seq- 基于相互作用组分析将确定神经病理性疼痛急性和慢性阶段的神经元-胶质细胞相互作用并确定药物靶向分子因素。目标 2 将确定神经元-神经胶质-神经元的行为意义利用不同细胞的化学遗传学激活和抑制，在神经病理性疼痛的不同阶段在 CeA 中发出信号类型和药理学（或基于病毒的）工具来调节分子因素。感觉阈值、情绪将测量反应、非诱发性持续疼痛以及焦虑和抑郁样行为。目标 3 将决定神经病理性疼痛不同阶段CeA中神经元-胶质细胞-神经元信号传导的电生理机制，使用脑切片中不同细胞类型的膜片钳记录以及化学遗传学和药理学（或病毒）基于）的操纵。成功完成这些概念性创新研究将显着推进我们的研究了解大脑可塑性和从急性到慢性疼痛转变中的神经免疫信号传导，并提供减轻慢性神经性疼痛的新目标。