Project #1 Single-soma RNA-seq and spatial transcriptomics of human TGs

项目

• 批准号: 10806547
• 负责人: Wenqin Luo
• 金额: $ 275.49万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10806547
• 关键词: Affect; Afferent Neurons; Atlases; Awareness; Biological Markers; Biology; Cell Nucleus; Cells; Collaborations; Complex; Data; Data Analyses; Data Set; Diagnosis; Dissociation; Drug Targeting; Epigenetic Process; Esthesia; Evaluation; Facial Pain; Foundations; Frozen Sections; Genes; Head; Headache; Headache Disorders; Helping to End Addiction Long-term; Household; Human; Individual; International; Knowledge; Mediating; Messenger RNA; Methods; Migraine; Molecular; Molecular Profiling; Neuroglia; Neurons; Pain; Pain Clinics; Pathologic; Pattern; Persons; Physiological; Protocols documentation; Publishing; Reporting; Research Personnel; Resolution; Sampling; Sensory; Spatial Distribution; Spinal Ganglia; Structure of trigeminal ganglion; System; Techniques; Tissues; Transcript; Trigeminal System; allodynia; burden of illness; cell type; chronic painful condition; deep sequencing; effective therapy; ganglion cell; human RNA sequencing; in vivo; laser capture microdissection; migraine treatment; multidisciplinary; multiple omics; nervous system disorder; neuronal cell body; novel; novel strategies; response; single nucleus RNA-sequencing; single-cell RNA sequencing; somatosensory; transcriptome; transcriptome sequencing; transcriptomics

Migraine, one of the most common primary headache disorders, affects 1 in 4 US households. This complex neurologic disorder is mediated in part by alterations in trigeminal somatosensation, which manifests as head/fa- cial pain and/or trigeminal allodynia. Effective treatments for migraine are still limited, and our knowledge about human trigeminal system at baseline and migraine conditions are sparse. In response to RFA-NS-22-018, HEAL Initiative: Discovery and Functional Evaluation of Human Pain-associated Genes & Cells, we propose to form the Penn Human Precision Pain Center (Penn HPPC) to elucidate molecular, cellular, epigenetic, and physiological profiles of human trigeminal ganglion (TG) sensory neurons at baseline and migraine conditions. The Penn HPPC will be composed of Penn and international investigators with multidisciplinary expertise. The PI, two MPIs, and two co-Is are currently collaborating on a single-soma deep RNA-seq of human dorsal root ganglion (DRG) neuron project, which form a strong foundation for this application. The Penn HPPC will contain three cores and perform three projects. The project 1 will be led by the PI, Dr. Luo, in collaboration with the Co-I, Dr. Ernfors, and supported by Dr. Li, PI of the data core, and Dr. Wu, PI of the project 2. Specifically, the project 1 will conduct single-soma RNAseq of neurons and non-neuron cells of human TGs from control donors and those with migraine. Three complementary approaches, a laser capture microdissection based single-neu- ron-soma deep RNA-seq (a novel method developed by the PI’s lab, which has been successfully used with human DRG neurons), 10 x Visium (a commercially available spatial transcriptomics platform), and MERSCOPE (another commercially available spatial transcriptomics platform) will be employed for this purpose. Aim 1 will focus on control human TG tissues, whereas Aim 2 will focus on human TG tissues from “migraine” donors. We will perform cell type deconvolution by integrating single-soma and 10x Visium datasets to generate human TG atlases with spatially-located single-soma transcriptomes of neurons and non-neuronal cells under normal and migraine conditions. We will also compare datasets between control and migraine donors. Taken together, our anticipated results will generate unprecedent molecular and cellular atlas of human TGs to understand normal trigeminal sensations and abnormal sensations associated with migraine. Our results may also lead to discovery of new biomarkers for migraine diagnosis and/or identification of novel potential drug targets for migraine treat- ment.

偏头痛是最常见的原发性头痛疾病之一，影响四分之一的美国家庭。这个复杂 神经障碍部分由三叉神经躯体感觉的改变介导，三叉神经躯体感觉的改变表现为头/发- 坐骨神经痛和/或三叉神经异常性疼痛。偏头痛的有效治疗方法仍然有限，我们对偏头痛的认识 人三叉神经系统在基线和偏头痛条件下是稀疏的。响应RFA-NS-22-018，HEAL 计划：人类疼痛相关基因和细胞的发现和功能评价，我们建议 成立宾州人类精确疼痛中心（Penn HPPC），以阐明分子，细胞，表观遗传， 在基线和偏头痛条件下的人三叉神经节（TG）感觉神经元的生理概况。 Penn HPPC将由具有多学科专业知识的Penn和国际研究人员组成。的 PI，两个MPI和两个co-I目前正在合作进行人类背根的单细胞体深层RNA测序 神经节（DRG）神经元项目，这为该应用奠定了坚实的基础。Penn HPPC将包含 三个核心，开展三个项目。项目1将由主要研究者罗博士领导， 共同研究员，Dr. Ernfors，并得到Data Core PI Li博士和项目PI Wu博士的支持2.具体而言是 项目1将对来自对照供体的人TG的神经元和非神经元细胞进行单胞体RNAseq 和偏头痛患者。三种互补的方法，基于激光捕获显微切割的单神经元， ron-soma deep RNA-seq（一种由PI实验室开发的新方法，已成功用于 人DRG神经元）、10 x Visium（市售的空间转录组学平台）和MERSCOPE （另一种市售的空间转录组学平台）将用于该目的。目标1将 Aim 2集中于对照人TG组织，而Aim 2集中于来自“偏头痛”供体的人TG组织。我们 将通过整合单个体细胞和10 x Visium数据集来执行细胞类型去卷积，以生成人TG 在正常条件下，神经元和非神经元细胞的空间定位单胞体转录组图谱， 偏头痛症状我们还将比较对照组和偏头痛供体之间的数据集。总的来说，我们的 预期的结果将产生前所未有的人类TG的分子和细胞图谱，以了解正常的 三叉神经感觉和与偏头痛相关的异常感觉。我们的研究结果也可能导致发现 用于偏头痛诊断的新生物标志物和/或用于偏头痛治疗的新的潜在药物靶点的鉴定- 我是说。