Project 1: Multi-omic characterization of human nociceptors

项目概要（项目1） 小鼠和人类疼痛治疗之间缺乏翻译，突出了动物和人类疼痛治疗的局限性。 疼痛模型和用来描述它们的分子工具。这促使人们做出重大努力， 这些分子特征可能为疼痛治疗提供新的思路 设计单细胞基因组学的最新进展现在使得表征基因表达谱成为可能， 空间位置，以及复杂组织中单个细胞的生理学，分辨率前所未有。我们 因此开发了用于表征人类伤害感受器的这些多维特征的方案， 并建议在此建立这些协议，并与其他U19 PRECISION人类疼痛中心。 分子 然后，我们建议利用我们的单细胞基因组和表观基因组方案来构建人类伤害感受器细胞 图谱来自不同的供体样本组。这些数据将有助于描述人类的多样性 伤害感受器亚型，建立这些亚型的表观基因组元件，以及常见遗传因素的影响， 这些临床上重要的细胞类型内基因表达的变异。这些数据集将提供一个 丰富的资源，用于挖掘新的疼痛治疗靶点，并解释常见的遗传变异如何影响 人类伤害感受器的基因表达。后一点对于解释病例对照研究尤其重要 疼痛研究界的研究。 结构 我们将生成并分析人类神经节的大规模单细胞空间转录组数据，以研究 伤害感受器亚型在感觉神经节中的位置以及在感觉神经节中是否存在独特的基因表达谱。 与其他神经元细胞相比，非神经元细胞（例如卫星神经胶质细胞）与伤害感受器密切相关 亚型 生理 我们将培养来自每个供体的新鲜感觉神经元，并测量它们的自发和辣椒素诱发的神经元。 活动然后，我们将使用单细胞空间转录组学来关联每个神经元中的基因表达谱 与其相关的自发和辣椒素诱发的活动。这些数据将生成一个资源， 整合分子和功能维度的伤害感受器。

PROJECT SUMMARY (Project 1) The lack of translation between mouse and human pain treatments has highlighted limitations of both the animal models of pain and molecular tools used to characterize them. This has prompted major efforts to characterize molecular features that are expressed in human nociceptors which may provide new ideas for pain therapeutic design. Recent advances in single-cell genomics now make it possible to characterize gene expression profiles, location in space, and the physiology of individual cells within complex tissues at unprecedented resolution. We have thus developed protocols for characterizing these multi-dimensional features of human nociceptors at single-cell resolution and propose to build upon these protocols here and in coordination with other U19 PRECISION Human Pain Centers. Molecular We then propose to leverage our single-cell genomic and epigenomic protocols to build a human nociceptor cell atlas derived from a diverse group of donor samples. These data will help characterize the diversity of human nociceptor subtypes, the epigenomic elements that establish these subtypes, and the effect of common genetic variation on gene expression within these clinically important cell types. Together, these datasets will provide a rich resource for mining novel pain therapeutic targets and for interpreting how common genetic variation affects gene expression in human nociceptors. This latter point is especially important for interpreting case-control studies across the pain research community. Structural We will generate and analyze large-scale single-cell spatial transcriptomic data of human ganglia to study the location of nociceptor subtypes in sensory ganglia and whether there are unique gene expression profiles in the non-neuronal cells (e.g. satellite glia) that are closely associated nociceptors as compared to other neuronal subtypes. Physiological We will culture fresh sensory neurons from each donor and measure their spontaneous and capsaicin-evoked activity. We will then use single-cell spatial transcriptomics to correlate gene expression profiles in each neuron with their associated spontaneous and capsaicin-evoked activity. These data will generate a resource that integrates both molecular and functional dimensions nociceptors.