基于单细胞核转录组测序分析面肩肱型肌营养不良症中细胞间通讯模式

Analysis of intercellular communication patterns in facioscapulohumeral muscular dystrophy based on single-cell nuclear transcriptome sequencing

目的:通过单细胞核转录组测序分析面肩肱型肌营养不良症(FSHD)中细胞间通讯模式。方法:选取2例FSHD患者的双侧不对称病变的口轮匝肌组织和2例正常口轮匝肌组织,共6例样本,分为对照组、轻度组和重度组。对照组为2例健康人的正常肌肉组织,轻度组和重度组分别为FSHD患者相对正常和损伤较重的一侧肌肉组织。对3组样本的全部细胞进行单细胞核转录组测序,鉴定差异表达基因和富集通路,并通过细胞通讯分析主要细胞类型间的细胞间通讯模式以及关键信号通路。结果:FSHD患者双侧肌肉样本的差异基因表达分析共鉴定了不同细胞类型中与疾病相关的46个功能性差异表达基因,与细胞凋亡、氧化应激、免疫炎症和肌肉功能等相关。FSHD重度组的细胞间通讯普遍增加。纤维/脂肪祖细胞(FAPs)和巨噬细胞是FSHD异常肌肉微环境中的重要信号来源,与疾病进展密切相关。FSHD组中存在6条独有信号通路:骨形态发生蛋白(BMP)、转化生长因子-β(TGF-β)、CXC基序趋化因子配体(CXCL)、黏附G蛋白偶联受体E5(ADGRE5)、白细胞介素-16(IL-16)和无翅型MMTV整合位点家族(WNT)信号通路,这些信号通路主要涉及巨噬细胞、FAPs和脂肪细胞间的相互作用,可能参与调控病变肌肉的脂肪沉积和纤维化改变。结论:单细胞核转录组测序提供了FSHD关键细胞类型间较为全面的细胞间通讯模式,为理解FSHD肌肉微环境的细胞间调控机制提供参考。

ObjectiveTo analyze the patterns of intercellular communication in facioscapulohumeral muscular dystrophy(FSHD)by single-cell nuclear transcriptome sequencing.MethodsBilateral asymmetrical lesions mouth orbicular muscle of two patients with FSHD and mouth orbicular muscle of two healthy patients were selected.Six samples were obtained,and were divided into control group,mild group and severe group.The normal orbicularis muscle sample was collected from 2 healthy individuals(the control group).The muscle samples in the mild group were from two patients with relatively normal muscle sides,and the samples in the severe group were from two patients with more severe muscle damage sides.Single-cell nuclear transcriptome sequencing was performed on all cells of the three groups.Reduced dimension clustering and cell definition were performed to identify differentially expressed genes and enrichment pathways.Intercellular communication patterns among major cell types and key signaling pathways were explored by cellular communication analysis.ResultsDifferential gene expression analysis of FSHD bilateral muscle samples identified 46 functionally differentially expressed genes associated with the disease in different cell types,related to apoptosis,oxidative stress,immune inflammation,and muscle function.Intercellular communication was generally increased in the severe group.Fibro-adipogenic progenitors(FAPs)and macrophages are important signaling sources in the abnormal muscle microenvironment of FSHD and are closely associated with disease progression.There are six unique signaling pathways in the mild group,including bone morphogenetic proteins(BMP),transforming growth factor-β(TGF-β),CXC motif chemokine ligand(CXCL),adhesion G protein-coupled receptor E5(ADGRE5),interleukin-16(IL-16),and wingless-type MMTV integration site family(WNT)signaling pathways.These signaling pathways are mainly involved in the interaction between macrophages,FAPs,and adipocytes and may be involved in the regulation of fat deposition and fibrosis changes in the diseased muscle.ConclusionsSingle-cell nuclear transcriptome sequencing provides a relatively comprehensive pattern of intercellular communication between key cell types in FSHD,providing an appropriate reference for understanding the intercellular regulatory mechanisms of the FSHD muscle microenvironment.