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

宾夕法尼亚大学人类精准疼痛中心 (HPPC)：正常和慢性疼痛条件下人类初级体感神经元类型的发现和功能评估

• 批准号: 10806545
• 负责人: Mingyao Li
• 金额: $ 675.15万
• 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/10806545
• 关键词: Affect; Afferent Neurons; Atlases; Biological Assay; Biological Markers; Biology; Cell Nucleus; Cells; Chromatin; Collaborations; Complex; Cryoultramicrotomy; DNA Methylation; Data; Data Analyses; Data Set; Diagnosis; Drug Targeting; Epigenetic Process; Esthesia; Evaluation; Facial Pain; Foundations; Frozen Sections; Gene Expression Profiling; Genes; Genomics; Harvest; Head; Headache Disorders; Helping to End Addiction Long-term; Household; Human; Individual; International; Knowledge; Link; Mediating; Messenger RNA; Methods; Methylation; Migraine; Molecular; Molecular Profiling; Nerve; Neuroglia; Neurons; Neurosurgeon; Nuclear; Pain; Pain Clinics; Patients; Persons; Phenotype; Physiological; Protocols documentation; Questionnaires; RNA; Research Personnel; Research Project Grants; Resolution; Sensory; Sorting; Spinal; Spinal Ganglia; Spinal nerve structure; Stains; Structure of trigeminal ganglion; System; Techniques; Testing; Transcript; Trigeminal System; Universities; Vertebral column; XCL1 gene; allodynia; blink reflexes; cell type; chronic painful condition; deep sequencing; effective therapy; epigenomics; human RNA sequencing; human data; human subject; human tissue; in vivo; laser capture microdissection; migraine treatment; multidimensional data; multidisciplinary; multiple omics; nervous system disorder; neuronal cell body; novel; novel strategies; recruit; response; 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. 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.

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，为该应用奠定了坚实的基础，特别是 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。 宾夕法尼亚州 HPPC 的管理实体将作为唯一的采购实体。 存储人体组织以及在研究项目之间分配人体组织的数据核心将是。 存储、处理和分发 HPPC 项目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，一种基于激光捕获显微切割的单神经元体深部 RNA-seq（由 PI 实验室开发的一种新方法，已成功用于人类 DRG neu- rons）、10 x Visium（一种市售的空间转录组学平台）和 MERSCOPE（另一个 com- 商业上可用的空间转录组学平台）进行神经元和非神经元的单体RNA测序 在项目 2 中，我们将进行两种类型的单细胞实验。 使用对照供体和偏头痛供体的 TG 进行细胞核多组学测序：染色质可及性 （ATAC 加 RNA，10x Genomics 多组）和 DNA 甲基化（甲基测定加 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、躯体- 感觉测试、眨眼反射以及三叉神经和脊髓感觉传入的体内显微神经造影记录。 我们提议的 Penn HPPC 将共同生成全面、多维的人类 TG 数据集 在基线和偏头痛条件下，这将生成前所未有的分子、细胞和功能图谱 了解正常的三叉神经感觉和与偏头痛相关的异常感觉。 还导致发现用于偏头痛诊断的新生物标志物和/或鉴定新型潜在药物 偏头痛治疗的目标。