The Penn Human Precision Pain Center (HPPC): Discovery and Functional Evaluation of Human Primary Somatosensory Neuron Types at Normal and Chronic Pain Conditions

偏头痛是最常见的原发性头痛疾病之一，每四个美国家庭中就有一个受到影响。这个建筑群 神经功能障碍的部分原因是三叉神经躯体感觉的改变，表现为头/脸-头-面-头-面-头-头-面-面 三叉神经痛和/或三叉神经痛。偏头痛的有效治疗方法仍然有限，我们对 人类三叉神经系统在基线和偏头痛的情况下是稀疏的。响应RFA-NS-22-018，修复 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 成立宾夕法尼亚人类精确疼痛中心(Penn HPPC)，以阐明分子、细胞、表观遗传学和 人三叉神经节(TG)感觉神经元在基线和偏头痛状态下的生理特征。 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调查人员组成。这个 PI、两个MPI和两个协同IS目前正在合作研究人类背根的单胞体深度RNA序列 神经节(DRG)神经元项目，为这一应用奠定了坚实的基础。具体地说，宾夕法尼亚大学的HPPC 将包含三个核心，并执行三个项目。行政核心将作为唯一的组织 和宾夕法尼亚HPPC的行政实体。人体组织核心将作为唯一的实体进行采购 以及存储人体组织并在研究项目中分配人体组织。数据核心将是 存储、处理和分发来自HPPC项目的所有数据的唯一实体。在项目1中，我们将 采用三种互补的方法：基于激光捕获显微切割的单个神经元胞体深度 RNA-seq(由PI实验室开发的一种新方法，已成功地用于人DRG-Neu. Rons)、10x Viimum(一种商用的空间转录学平台)和MERSCOPE(另一种COM- 市场上可用的空间转录平台)，以进行神经元和非神经元的单体体RNA序列 对照供者和偏头痛患者的TGS集落细胞。在项目2中，我们将执行两种类型的单人- 用对照供体和偏头痛患者的TGS进行核多组学测序：染色质的可及性 (ATAC+RNA，10倍基因组多组分析)和DNA甲基化(甲基化+RNA，snmCAT-seq)。 在项目3中，我们将招募偏头痛患者和对照受试者进行疼痛问卷调查，SOMA- 对三叉神经和脊髓感觉传入进行感觉测试、瞬目反射和体内显微神经学记录。 总之，我们建议的宾夕法尼亚大学HPPC将产生人类TGS的全面和多维数据集 在基线和偏头痛条件下，这将产生前所未有的分子、细胞和功能图谱 了解与偏头痛相关的正常三叉神经感觉和异常感觉。我们的结果可能 也导致发现用于偏头痛诊断和/或识别新的潜在药物的新的生物标志物 偏头痛治疗的靶点。

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. Specifically, the Penn HPPC will contain three cores and perform three projects. The administrative core will serve as a sole organizational and administrative entity for the Penn HPPC. The human tissue core will function as the sole entity for procuring and storing human tissues and distributing human tissues among research projects. The data core will be the sole entity for storage, processing, and distribution of all data from the HPPC projects. In project 1, we will employ three complementary approaches, a laser capture microdissection based single-neuron-soma deep RNA-seq (a novel method developed by the PI’s lab, which has been successfully used with human DRG neu- rons), 10 x Visium (a commercially available spatial transcriptomics platform), and MERSCOPE (another com- mercially available spatial transcriptomics platform) to conduct single-soma RNA-seq of neurons and non-neu- ronal cells of TGs from control donors and those with migraine. In project 2, we will perform two types of single- nucleus multi-omics sequencing with TGs from control donors and those with migraine: chromatin accessibility (ATAC plus RNA, 10x Genomics multiome assay) and DNA methylation (methylation plus RNA, snmCAT-seq). In project 3, we will recruit migraine patients and control human subjects to conduct pain questionnaires, soma- tosensory tests, blink reflex, and in vivo microneurography recordings of trigeminal and spinal sensory afferents. Together, our proposed Penn HPPC will produce comprehensive and multi-dimensional datasets of human TGs at baseline and migraine conditions, which will generate unprecedent molecular, cellular, and functional atlas 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 treatment.