Project #2 Integrated single-nucleus multi-omics (ATAC-seq+RNA-seq or chromatin accessibility + RNA-seq) of human TGs

偏头痛是一种流行的原发性头痛疾病，在美国每四个家庭中就有一个受到影响。 美国。这种复杂的神经障碍在一定程度上是由三叉神经躯体感觉的改变所介导的， 表现为头部和面部疼痛和/或三叉神经痛。尽管它很流行，但目前有 偏头痛的有效治疗方法有限，以及我们对人类三叉神经系统的了解 基线和偏头痛的情况很稀少。响应RFA-NS-22-018，修复倡议：发现和 人类疼痛相关基因和细胞的功能评估，我们建议形成宾夕法尼亚人 精确疼痛中心(Penn HPPC)专注于阐明分子、细胞、表观遗传学和生理学- 人三叉神经节(TG)的形态特征，它含有支配大脑皮质的初级体感神经元。 头部感觉，在基线和偏头痛条件下。宾夕法尼亚大学的HPPC将由宾夕法尼亚大学和国际米兰大学组成。 拥有多学科专业知识的国家调查人员。PI、两个协同PI和两个协同IS目前正在合作- 在人背根神经节(DRG)神经元的单个胞体深RNA序列上，形成了一个强大的 此应用程序的基础。宾夕法尼亚大学的氟氯烃淘汰管理方案将包含三个核心，并执行三个项目。郝伟博士 吴是一位神经表观遗传学和单细胞基因组学专家，他将在明耀博士的支持下领导项目2 LI，数据核心的PI。该项目旨在使用两种单核多组学分析方法，包括： 高度平行的10倍基因组多组分析(3‘偏向的mRNA和开放染色质)和SnmCAT-seq(全 长度mRNA、开放染色质和DNA甲基化)，以全面定位转录和表观基因组 (染色质可及性和/或DNA甲基化)在对照供体和受试者中的单核分辨率 偏头痛。在项目2中，目标1将专注于控制三叉神经节(TG)组织，而目标2将专注于 偏头痛捐献者的TG组织。预期的结果将产生全面的表观遗传学和 转录图谱，以了解与偏头痛相关的正常和异常三叉神经感觉。这 丰富的资源也可能导致发现偏头痛诊断的新生物标志物和新的潜在药物 偏头痛治疗的靶点。

Migraine is a prevalent primary headache disorder that impacts one out of every four households in the United States. This complex neurologic disorder is mediated in part by alterations in trigeminal somatosensation, which manifests as head and facial pain and/or trigeminal allodynia. Despite its prevalence, there are currently limited effective treatments available for migraine, and our knowledge about human trigeminal system under 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 focus on elucidating molecular, cellular, epigenetic, and physiolog- ical profiles of human trigeminal ganglion (TG), which contains primary somatosensory neurons governing the head sensation, under baseline and migraine conditions. The Penn HPPC will be composed of Penn and inter- national investigators with multidisciplinary expertise. The PI, two co-PIs, and two co-Is are currently collaborat- ing 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. Dr. Hao Wu, an expert in neuroepigenetics and single-cell genomics, will lead project 2 with support from Dr. Mingyao Li, the PI of the data core. This project aims to use two single-nucleus multi-omics assays, including the mas- sively parallel 10x Genomics multiome assay (3’ biased mRNA and open chromatin) and snmCAT-seq (full- length mRNA, open chromatin, and DNA methylation), to comprehensively map transcriptomic and epigenomic (chromatin accessibility and/or DNA methylation) profiles at single-nucleus resolution in control donors and those with migraine. In project 2, Aim 1 will focus on control trigeminal ganglion (TG) tissues, while Aim 2 will focus on TG tissues from "migraine" donors. The anticipated results will produce a comprehensive epigenetic and transcriptomic atlas to understand normal and abnormal trigeminal sensations associated with migraine. This rich resource may also lead to the discovery of new biomarkers for migraine diagnosis and novel potential drug targets for migraine treatment.