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

项目摘要-项目1 慢性疼痛困扰着超过1亿美国人，是寻求医疗的主要原因。一些新 在过去的几十年里，已经提供了治疗，导致阿片类药物的过度使用，尽管它们有可能 在许多慢性疼痛病症中的滥用和有限的功效。一个突出的问题是， 临床前模型，以产生有效的疼痛治疗剂。解决这一问题的一个关键障碍是 我们对人类周围神经系统的理解和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，我们建议确定参与的其他基因， 通过分析一组特发性疼痛性神经病患者的基因组，研究人类疼痛通路。 虽然伤害性神经元是外周疼痛传递最关键的成分，但外周神经元中的其他细胞也是如此。 周围神经--神经胶质细胞和免疫细胞，特别是巨噬细胞--是疼痛的重要调节器 这些神经元的反应。需要进一步分析，以了解这些细胞之间的相互作用， 神经成分和感觉轴突导致疼痛。因此，我们建议使用单核 我们最近利用RNAseq策略来产生小鼠外周神经图谱， 人类周围神经细胞成分表达谱的综合图谱。 在这个建议中，我们概述了一系列的实验，以检查基因表达在细胞中的人类 外周神经和鉴定与特发性疼痛性神经病相关的新基因变异。整个 基因组测序将对来自一个大的具有良好特征的患有疼痛的患者队列的样本进行。 神经病变基因变异将与表型信息相关联，以识别参与基因突变的新基因。 疼痛通路使用多组学技术，包括snRNA-seq和snATAC-seq以及空间 转录组学和多重蛋白质组学，从我们的供体组织核心研究神经，我们将设计一个 人外周血淋巴细胞基因表达和细胞组成的综合性、无偏倚单核图谱 神经将进行平行研究，以确定细胞成分和基因表达模式， 手术切除的疼痛性神经瘤通过比较神经瘤转录组和正常组织的图谱， 神经数据集，我们计划确定疼痛神经瘤的细胞类型特异性决定因素。我们期待这些富有的， 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 疼痛综合征的研究，这将为疼痛的治疗方案提供见解。

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.