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

多组学周围神经图谱能够精细绘制疼痛分子表型

基本信息

批准号：
10707409
负责人：
JEFFREY D MILBRANDT
金额：
$ 72.78万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-30 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10707409
关键词：
Address American Atlases Axon Candidate Disease Gene Cell Nucleus Cells Chromatin Clinical Communities Cutaneous Data Data Set Disease Failure Femur Foundations Functional disorder Gene Combinations Gene Expression Gene Expression Profile Genes Genetic Transcription Genome Genomic approach Genomics Genotype Goals Grain Human Immune Knowledge Lateral Macrophage Maps Medical Methods Molecular Motor Mus Mutation Nerve Neuroglia Neurologic Neuroma Neurons Nociceptors Ontology Operative Surgical Procedures Organ Donor Pain Pain Map Pathway Analysis Pathway interactions Patients Pattern Peripheral Nerves Peripheral Nervous System Phenotype Pre-Clinical Model Proteomics RNA Research Resected Resolution Resources Rodent Sampling Schwann Cells Sensory Series Statistical Data Interpretation Syndrome Technology Therapeutic Tissue Donors Tissue Procurements Tissues Traumatic injury Variant XCL1 gene abuse liability afferent nerve axonal sprouting candidate validation cell type chronic pain chronic painful condition cohort data integration experimental study gene discovery genetic variant genome analysis genome sequencing insight molecular phenotype multiple omics neuron component novel novel therapeutics opioid overuse pain model painful neuropathy patient stratification peripheral pain pre-clinical response risk variant single nucleus RNA-sequencing transcriptome transcriptome sequencing transcriptomics transmission process whole genome

项目摘要

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.

项目摘要 - 项目1 慢性疼痛遭受了超过1亿美国人的困扰，是寻求医疗的主要原因。很少有新在过去的几十年中，疗法已经进行了，导致阿片类药物过度使用，dospite的潜力在许多慢性疼痛状况下滥用和有限的效率。一个明显的关注是研究的失败临床前模型，以产生有效的疼痛疗法的药物。解决此问题的关键障碍是我们对人类外周神经系统的理解与研究之间的知识差距在啮齿动物的临床前疼痛模型中进行。在这个项目中，我们建议确定涉及的其他基因通过分析特发性疼痛神经病患者的基因组的人类疼痛途径。伤害性神经元是外围疼痛传播最重要的组成部分，但其他细胞中的其他细胞周围神经 - glia和免疫细胞，尤其是巨噬细胞 - 是疼痛的重要调节剂这些神经元中的反应。需要进行其他分析以了解这些细胞之间的相互作用如何神经成分和感觉轴突有助于疼痛。因此，我们建议使用单核我们最近使用的RNASEQ策略来产生小鼠外周神经地图集，以创建多层，人周围神经细胞成分的表达谱的全面地图集。在此提案中，我们概述了一系列检查人类细胞中基因表达的实验周围神经并确定与特发性疼痛神经病相关的新型基因变异。所有的基因组测序将对来自大量疼痛的大量特征良好患者的样品进行神经病。基因变异将与表型信息相关，以识别涉及的新基因疼痛途径。使用包括SNRNA-SEQ和SNATAC-SEQ在内的多摩变技术以及空间转录组学和多重蛋白质组学，为了从我们的供体组织核心研究神经，我们将设计一个人类周围的基因表达和细胞组成的全面，公正的单核图谱神经。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神经瘤。通过将神经瘤转录组与源自正常的地图集进行比较神经数据集，我们计划识别疼痛神经瘤的细胞类型特异性决定剂。我们期望这些富人完全数据集为感觉神经功能障碍的分子和细胞分析提供基础在疼痛综合征中，可以洞悉疼痛的治疗选择。