Multi-omics peripheral nerve atlas enables fine-mapping of pain molecular phenotypes

项目摘要-项目1 慢性疼痛困扰着1亿多美国人，是人们寻求治疗的主要原因。几乎没有新的东西 在过去的几十年里，已经提供了治疗方法，导致过度使用阿片类药物，尽管它们有可能 在许多慢性疼痛情况下，滥用和疗效有限。一个值得关注的问题是，使用 临床前模型，以产生有效的止痛药物。解决这个问题的一个关键障碍是 我们对人类周围神经系统的了解与研究之间的知识差距 在啮齿动物临床前疼痛模型中进行。在这个项目中，我们建议识别更多与 通过对一组特发性痛性神经病患者的基因组分析，研究人类的痛觉通路。 虽然伤害性神经元是外周疼痛传递最关键的组成部分，但在 周围神经--神经胶质细胞和免疫细胞，尤其是巨噬细胞--是疼痛的重要调节细胞。 这些神经元的反应。需要进一步的分析才能理解这些细胞之间的相互作用 神经成分和感觉神经轴突对疼痛起作用。因此，我们建议使用单核 RNAseq策略，我们最近利用制作的小鼠周围神经图谱来创建多层， 人类周围神经细胞成分表达谱的综合图谱。 在这项提案中，我们概述了一系列实验，以检测人类细胞中的基因表达 并确定与特发性痛性神经病相关的新的基因变异。整体 基因组测序将对一大批特征良好的疼痛患者的样本进行。 神经病。基因变异将与表型信息相关联，以识别参与 疼痛路径。使用多组学技术，包括SNRNA-seq和snatac-seq以及空间 转录组学和多重蛋白质组学，为了研究我们供体组织核心的神经，我们将设计一种 全面、无偏见的人类外周血细胞基因表达和细胞组成的单核图谱 太紧张了。将进行平行研究，以确定细胞成分和基因表达模式 手术切除了疼痛的神经瘤。通过将神经瘤转录本与正常来源的图谱进行比较 神经数据集，我们计划确定疼痛神经瘤的细胞类型特定决定因素。我们希望这些富有的和 补充数据集，为感觉神经功能障碍的分子和细胞分析提供基础 这将为疼痛的治疗选择提供见解。

Project Abstract – Project 1 Chronic pain afflicts over 100 million Americans and is a leading reason for seeking medical treatment. Few new therapies have been delivered in the past decades, resulting in overuse of opioids, despite their potential for abuse and limited efficacy in many chronic pain conditions. One glaring concern is the failure of studies using preclinical models to yield agents that are effective pain therapeutics. A key barrier in addressing this problem is the knowledge gap between our understanding of the human peripheral nervous system and the studies performed in rodent preclinical pain models. In this project, we propose to identify additional genes involved in human pain pathways via analysis of the genomes of a cohort of patients with idiopathic painful neuropathy. While nociceptive neurons are the components most critical for peripheral pain transmission, other cells in peripheral nerves—glia and immune cells, particularly macrophages—are important regulators of pain responses in these neurons. Additional analysis is required to understand how interactions among these cellular nerve components, and with sensory axons, contribute to pain. We therefore propose to use single-nuclei RNAseq strategies that we recently utilized to produce a mouse peripheral nerve atlas to create a multi-layered, comprehensive atlas of the expression profiles of human peripheral nerve cellular components. In this proposal, we outline a series of experiments to examine gene expression in cells of human peripheral nerve and to identify novel gene variants associated with idiopathic painful neuropathy. Whole genome sequencing will be performed on samples from a large cohort of well-characterized patients with painful neuropathy. Gene variants will be associated with phenotypic information to identify new genes involved in the pain pathway. Using multi-omic technologies, including snRNA-seq and snATAC-seq as well as spatial transcriptomics and multiplex proteomics, to study nerves from our donor tissue core, we will devise a comprehensive, unbiased single-nuclei atlas of gene expression and cellular composition of human peripheral nerves. Parallel studies will be performed to determine the cellular components and gene expression patterns of surgically resected painful neuromas. By comparing the neuroma transcriptomes to the atlas derived from normal nerve datasets, we plan to identify cell type-specific determinants of painful neuromas. We expect these rich and complementary datasets to provide a foundation for molecular and cellular analysis of sensory nerve dysfunction in pain syndromes that will provide insights into treatment options for pain.