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，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，为该应用奠定了坚实的基础。具体来说，Penn 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机构， 宾夕法尼亚州HPPC的行政实体。人体组织核心将作为唯一的实体， 以及储存人体组织和在研究项目中分配人体组织。数据核心将是 唯一负责存储、处理和分发来自HPPC项目的所有数据的实体。在项目1中，我们 采用三种互补的方法，激光捕获显微切割基于单神经元-索马深 RNA-seq（PI实验室开发的一种新方法，已成功用于人DRG neu-1基因的研究） rons）、10 x Visium（一种商业上可用的空间转录组学平台）和MERSCOPE（另一种COM- 商业上可获得的空间转录组学平台）进行神经元和非神经元的单胞体RNA-seq。 对照组和偏头痛患者的TG的ronal细胞。在项目2中，我们将执行两种类型的单- 对照供体和偏头痛供体TG的核多组学测序：染色质可及性 (ATAC加RNA，10 x Genomics多基因组测定）和DNA甲基化（甲基化加RNA，snmCAT-seq）。 在项目3中，我们将招募偏头痛患者和对照人类受试者进行疼痛问卷调查，索马- tosensory测试，眨眼反射，并在体内三叉神经和脊髓感觉传入的显微神经造影记录。 总之，我们提出的Penn HPPC将产生全面和多维的人类TG数据集 这将产生前所未有的分子，细胞和功能图谱， 了解正常三叉神经感觉和与偏头痛相关的异常感觉。我们的研究结果可能 还导致发现用于偏头痛诊断和/或鉴定新的潜在药物的新生物标志物 治疗偏头痛的目标。

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.