Anatomic, Physiologic and Transcriptomic Mechanisms of Neuropathic Pain in Human DRG

人类背根神经节神经病理性疼痛的解剖学、生理学和转录组学机制

基本信息

批准号：
10268154
负责人：
Patrick M Dougherty
金额：
$ 14.95万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-15 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10268154
关键词：
Address Affect Agonist Anatomy Automobile Driving Bioinformatics Biological Biophysics Brain Cancer Center Cells Chest Code Data Data Set Dependence Electrophysiology (science)Epidemic Family Female G-Protein-Coupled Receptors Gene Expression Profile Generations Genes Genetic Transcription Goals Harvest Human Immune System Diseases Immunohistochemistry In Situ Hybridization Interleukin-6 Intervention Literature Modeling Molecular Neuroimmune Neuronal Dysfunction Neuronal Plasticity Neurons Neurosciences Neurosurgical Procedures Nociceptors Operative Surgical Procedures Pain Pain Research Pathway interactions Patients Pharmacology Phenotype Physiological Pre-Clinical Model Proliferating Publishing Purinergic P1 Receptors RNA Regulatory Pathway Sampling Signal Pathway Signal Transduction Societies Spinal Ganglia Sum Technology Testing Texas Therapeutic Time Tissues Translations Tumor-infiltrating immune cells Universities Untranslated RNA Up-Regulation Vertebral column Work base biomarker discovery case control cell type chronic pain clinical care clinical pain cohort dermatome experimental study follow-up human female insight jun Oncogene male member neuronal excitability neurophysiology novel oncostatin M pain model pain patient painful neuropathy patch clamp pre-clinical receptor relating to nervous system sex sexual dimorphism single-cell RNA sequencing therapeutic development transcription factor transcriptome transcriptome sequencing transcriptomics

项目摘要

We have launched a collaborative effort between MD Anderson Cancer Center and University of Texas at Dallas that uses dorsal root ganglion (DRG) removed from pain phenotyped patients during neurological surgery. DRGs are taken from thoracic levels during a spine stabilization surgery and then cut in thirds. One third is saved for immunohistochemistry (IHC) or in situ hybridization (ISH), one third goes for culturing and electrophysiology, and one third is used for RNA sequencing. We have developed an extensive dataset with patient pain phenotype information, DRG neuron electrophysiological characterization and RNA sequencing. In many cases we have pairs of DRGs from the same patient where the patient had pain in one dermatome and not in another, allowing for precise case-control analysis. Our electrophysiology results clearly demonstrate that chronic pain is associated with spontaneous activity (SA) in DRG nociceptors. This is the first time that this has been demonstrated. Our RNA sequencing results identify transcriptional changes associated with chronic pain and SA in the DRG that show indications of sexual dimorphism. In males we find clear signs of immune infiltration and neuro-immune interactions as well as an increase in expression for some members of the FOS/JUN transcription factor family. In females we see an upregulation of some G-protein coupled receptors (GPCRs) and other signs of intrinsic changes in neuronal excitability. These findings give unique insights into drivers of chronic pain in the DRG in a diverse cohort of patients with important implications for chronic pain therapeutic development, including the potential need for sex-specific treatment. Our overarching hypothesis is that SA in human DRG neurons, which is a critical factor for pain generation in patients, is driven by fundamentally different mechanisms in male and female patients. We will test this hypothesis using human DRG samples and a combination of electrophysiology (Aim 1) and RNA-seq (Aim 2). In Aim 3 IHC and ISH along with pharmacological interventions guided by preliminary findings and data generated during this project will be used to define new potential therapeutic avenues. In sum, the experiments in this project will give fundamental new insight into mechanisms of chronic pain that will enable therapeutic and biomarker discovery with the opportunity for an almost immediate impact on clinical care.

我们发起了 MD 安德森癌症中心和美国大学之间的合作项目德克萨斯州达拉斯市使用从疼痛表型患者身上切除的背根神经节 (DRG) 在神经外科手术期间。 DRG 在脊柱稳定过程中从胸部水平获取手术，然后切成三分之一。三分之一用于免疫组织化学 (IHC) 或原位检测杂交（ISH），三分之一用于培养和电生理学，三分之一用于 RNA测序。我们开发了包含患者疼痛表型的广泛数据集信息、DRG 神经元电生理特征和 RNA 测序。在许多在某些情况下，我们有来自同一患者的成对 DRG，其中患者的一侧出现疼痛皮刀而不是另一个，可以进行精确的病例对照分析。我们的电生理学结果清楚地表明，慢性疼痛与自发活动（SA）有关 DRG 伤害感受器。这是第一次得到证实。我们的RNA测序结果确定了与慢性疼痛和 DRG 中的 SA 相关的转录变化显示出性别二态性的迹象。在男性中，我们发现明显的免疫渗透迹象神经免疫相互作用以及某些成员的表达增加 FOS/JUN 转录因子家族。在女性中，我们发现某些 G 蛋白的上调耦合受体（GPCR）和神经元兴奋性内在变化的其他迹象。这些研究结果为不同群体中 DRG 慢性疼痛的驱动因素提供了独特的见解对慢性疼痛治疗发展具有重要影响的患者，包括潜在需要针对性别的治疗。我们的首要假设是人类 DRG 中的 SA 神经元是患者产生疼痛的关键因素，从根本上驱动男性和女性患者的机制不同。我们将用人类来检验这个假设 DRG 样本以及电生理学（目标 1）和 RNA-seq（目标 2）的组合。目标 3 IHC 和 ISH 以及以初步结果和数据为指导的药物干预该项目期间产生的成果将用于确定新的潜在治疗途径。总而言之，该项目的实验将为慢性病的机制提供基本的新见解疼痛将使治疗和生物标志物的发现成为可能对临床护理产生直接影响。