Dissecting myogenic-endothelial-immune interactomes in human ME/CFS skeletal muscles

剖析人类 ME/CFS 骨骼肌中的肌源性内皮免疫相互作用组

• 批准号: 10627290
• 负责人: Benjamin David Cosgrove
• 金额: $ 36.5万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10627290
• 关键词: ATAC-seq; Architecture; Atlases; Attention; Automobile Driving; Awareness; Bioinformatics; Biological Assay; Biological Markers; Biological Specimen Banks; Biopsy; Biopsy Specimen; Blood; Blood Vessels; Blood specimen; Cell Communication; Cell Culture Techniques; Cell Nucleus; Cells; Cellular Assay; Chronic Fatigue Syndrome; Collection; Coupled; Custom; Data; Data Set; Databases; Diagnosis; Disease; Endothelial Cells; Endothelium; Engineering; Epigenetic Process; Etiology; Exertion; Exposure to; Fatigue; Functional disorder; Gene Expression; Genetic Transcription; Hand Strength; Health; Histopathology; Homeostasis; Human; Immune; Immune System Diseases; Immunophenotyping; Individual; Ligands; Link; Maps; Mediator; Metabolic; Metabolic dysfunction; Methods; MicroRNAs; Modeling; Molecular; Muscle; Muscle Cells; Muscle Fatigue; Muscle Fibers; Myeloid Cells; Needles; Pathway interactions; Patients; Peripheral; Physiological; Physiology; Plasma; Prevalence; RNA; Regulation; Regulator Genes; Research; Resolution; Role; Sampling; Severities; Signal Pathway; Signal Transduction; Skeletal Muscle; Small Nuclear RNA; Surveys; Symptoms; System; Testing; Untranslated RNA; Urine; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelium; antagonist; biobank; biological sex; cell type; cohort; endothelial dysfunction; epigenome; epigenomics; human model; innovation; insight; intercellular communication; monocyte; multiple omics; muscle metabolism; neutralizing antibody; novel; programs; receptor; recruit; repaired; single cell technology; single nucleus RNA-sequencing; systemic inflammatory response; therapeutic target; transcriptome; transcriptome sequencing; transcriptomics

PROJECT 1: ABSTRACT/SUMMARY Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a heterogeneous debilitating disorder characterized by severe disabling fatigue, especially post-exertion, in the absence of alternative diagnosis. The etiology of ME/CFS remains unknown and underexplored and is likely multifaceted. Given the prevalence of muscle fatigue symptoms, the pathophysiology of skeletal muscle has received attention in ME/CFS studies. Skeletal muscles of ME/CFS individuals do not exhibit overt changes in muscle fiber architecture, quality, or type, though studies have identified altered muscle metabolism, peripheral vascular dysfunction, and local and systemic inflammation as potential mediators of CE/MFS. This project will address the premise that skeletal muscle dysfunction in ME/CFS arises from a multifaceted and heterogeneous dysregulation of myogenic, vascular endothelial, and immune cell identities and interactions in skeletal muscles. Myogenic cells and resident immune and vascular endothelial cells are in close proximity and have varied interactions through cell-cell signaling pathways and coupled metabolic programs that are critical for muscle health and disease. Here, we will collect a new cohort of ME/CFS and control skeletal muscle biopsies to investigate the interactions between myogenic, endothelial, and muscle-resident immune cells using innovative single-cell technologies. First, we will collect a new cohort of skeletal muscle biopsies and blood samples from ME/CFS patients (n = 40) and healthy controls (n = 20), along with accompanying survey and physiological data (grip strength and orthostatic vascular function). Building on our prior track record of generating a multi-donor scRNAseq atlas in human skeletal muscle, we will analyze these new biopsies by single-nucleus paired RNA/ATAC-sequencing (snRNA/ATACseq) and a novel method of spatial total RNA-sequencing (STRS). snRNA/ATACseq will reveal changes in cellular identities and compositions, as well as alterations in metabolic and transcriptional programs through various bioinformatic analyses. STRS will reveal changes in the spatial mapping of muscle cell types and their noncoding (e.g., regulatory miRNA and lncRNA) RNA profiles. Together these data will be integrated to identify changes in cell-cell communication (ligand-receptor) pathways involving myogenic, endothelial, and immune cells and their spatial organization between control and ME/CFS muscles. Various metrics from these single-cell datasets will be tested through association studies as possible biomarkers that may explain the heterogeneous manifestations of ME/CFS in this patient cohort. Third, hypotheses related to signaling interactions driving endothelial dysfunction in ME/CFS skeletal muscles will be evaluated using cell culture models involving skeletal muscle- specific human endothelial cells exposed to plasma and monocytes isolated from ME/CFS or control samples. Together, these aims may shed light on the cellular and molecular etiologies of ME/CFS in human skeletal muscles.

项目1：摘要/总结 肌痛性脑脊髓炎/慢性疲劳综合征（ME/CFS）是一种异质性的衰弱性疾病 在没有其他诊断的情况下，以严重的致残性疲劳为特征，尤其是劳累后。的 ME/CFS的病因学仍然未知，研究不足，可能是多方面的。鉴于普遍存在的 肌肉疲劳症状，骨骼肌的病理生理学在ME/CFS研究中受到关注。 ME/CFS个体的骨骼肌在肌纤维结构、质量或功能方面没有明显的变化。 型，虽然研究已经确定改变肌肉代谢，外周血管功能障碍，局部和 全身炎症是CE/MFS的潜在介质。这个项目将解决的前提是，骨骼 ME/CFS中的肌肉功能障碍源于肌源性的多方面和异质性的失调， 血管内皮细胞和免疫细胞的身份和骨骼肌中的相互作用。生肌细胞和常驻 免疫和血管内皮细胞非常接近，并通过细胞-细胞间相互作用而变化 信号通路和耦合的代谢程序，对肌肉健康和疾病至关重要。这里我们 将收集一组新的ME/CFS和对照骨骼肌活检样本，以研究ME/CFS和对照骨骼肌活检之间的相互作用。 肌源性、内皮和肌肉驻留免疫细胞使用创新的单细胞技术。一是 收集来自ME/CFS患者（n = 40）和健康人（n = 40）的骨骼肌活检和血液样品的新队列。 对照组（n = 20），沿着，并伴随调查和生理数据（握力和立位血管 函数）。基于我们先前在人类骨骼中生成多供体scRNAseq图谱的记录， 肌肉，我们将通过单核配对RNA/ATAC测序（snRNA/ATACseq）分析这些新的活检组织 以及一种新的空间全RNA测序方法（STRS）。snRNA/ATACseq将揭示细胞内的变化， 身份和组成，以及通过各种代谢和转录程序的改变， 生物信息学分析STRS将揭示肌肉细胞类型及其非编码空间映射的变化 (e.g.,调节miRNA和lncRNA）RNA谱。这些数据将被整合在一起，以确定 涉及肌原细胞、内皮细胞和免疫细胞的细胞-细胞通讯（配体-受体）途径及其 控制和ME/CFS肌肉之间的空间组织。这些单细胞数据集的各种指标将 通过相关研究进行测试，作为可能的生物标志物，可以解释异质性表现 ME/CFS的发病率。第三，与驱动内皮细胞的信号相互作用相关的假设， ME/CFS骨骼肌的功能障碍将使用涉及骨骼肌的细胞培养模型进行评估， 暴露于从ME/CFS或对照样品分离的血浆和单核细胞的特异性人内皮细胞。 总之，这些目标可能有助于阐明人类骨骼肌ME/CFS的细胞和分子病因学。 肌肉.