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，影响着四分之一的美国家庭。 神经系统疾病部分是由三叉神经躯体感觉的改变介导的，表现为头/fa- 偏头痛的有效治疗方法仍然有限，而且我们对偏头痛的了解仍然有限。 基线和偏头痛情况下的人类三叉神经系统稀疏 根据 RFA-NS-22-018，HEAL。 倡议：人类疼痛相关基因和细胞的发现和功能评估，我们建议 成立宾夕法尼亚州人类精准疼痛中心 (Penn HPPC)，以阐明分子、细胞、表观遗传和 人类三叉神经节 (TG) 感觉神经元在基线和偏头痛条件下的生理特征。 宾夕法尼亚大学 HPPC 将由宾夕法尼亚大学和具有多学科专业知识的国际研究人员组成。 PI、两个 MPI 和两个 co-Is 目前正在合作开展人类背根的单体细胞深层 RNA 测序 神经节 (DRG) 神经元项目，该项目为该应用奠定了坚实的基础。 项目 1 将由 PI 罗博士领导，并与 3 个项目合作。 Co-I，Ernfors 博士，并得到数据核心 PI 李博士和项目 2 PI 吴博士的支持。 项目1将对来自对照供体的人类TG的神经元和非神经元细胞进行单体RNAseq 以及偏头痛患者的三种互补方法，一种是基于激光捕获显微切割的单神经元治疗方法。 ron-soma Deep RNA-seq（PI实验室开发的一种新方法，已成功应用于 人类 DRG 神经元）、10 x Visium（商用空间转录组学平台）和 MERSCOPE （另一个市售的空间转录组学平台）将用于此目的。 重点关注对照人类 TG 组织，而 Aim 2 将重点关注来自“偏头痛”供体 We 的人类 TG 组织。 将通过集成单体细胞和 10x Visium 数据集来执行细胞类型反卷积以生成人类 TG 正常和非神经元细胞的空间定位单体转录组图谱 我们还将比较对照和偏头痛捐赠者之间的数据集。 预期结果将生成前所未有的人类 TG 分子和细胞图谱，以了解正常情况 我们的研究结果也可能会导致与偏头痛相关的三叉神经感觉和异常感觉的发现。 用于偏头痛诊断的新生物标志物和/或鉴定用于偏头痛治疗的新型潜在药物靶标 ment。