应用单细胞测序识别银屑病致病相关的潜在生物标志物以及细胞互作关系  

Identification of potential pathogenic biomarkers and cell interaction associated with psoriasis using single-cell sequencing

作　　者：陈东宇 胡敏 王朴 潘素跃 杨晓雨 黄巧 樊文龙 王红心 何玉清 Chen Dongyu;Hu Min;Wang Pu;Pan Suyue;Yang Xiaoyu;Huang Qiao;Fan Wenlong;Wang Hongxin;He Yuqing(Department of Epidemiology and Medical Statistics,Institute of Medical Systems Biology,Guangdong Medical University,Dongguan 523808,China;Dermatological Department,Liaobu Hospital,Dongguan 523400,China)

机构地区：[1]广东医科大学公共卫生学院流行病与卫生统计学系,广东医科大学医学系统生物学研究所,东莞523808 [2]东莞市寮步医院皮肤科,东莞523400

出　　处：《国际遗传学杂志》2023年第1期1-13,共13页International Journal of Genetics

基　　金：国家自然科学基金(81773312);广东省"扬帆计划引进紧缺拔尖"人才项目(201433005)。

摘　　要：目的在单细胞水平上探究与银屑病致病相关的潜在生物标志物和细胞间的异质性水平。方法对银屑病皮肤活检组织的单细胞测序数据进行质控、降维聚类及细胞亚群注释,以及对细胞类群进行差异基因(differentially expressed genes,DEGs)、细胞通讯及拟时序分析,并构建TFs-miRNA-hub基因调控网络。结果基于多算法共鉴定出6个共同交集的hub基因,分别为SPRR2A、SPRR2D、IL7R、IL1RN、IER3、LCN2。KEGG显示其主要涉及NF-κB信号通路、TNF信号通路、MAPK信号通路等。在内皮细胞、成纤维细胞以及免疫细胞如朗格汉斯细胞、CD8^(+)T细胞、M1巨噬细胞、静息T细胞里SPRRs基因表达量均显著上调,IL7R则主要在多种免疫细胞中过表达。成纤维细胞、树突状细胞、Treg细胞和CD8^(+)T细胞在银屑病皮损的含量显著增多(P=0.023、P=0.007、P=0.046、P=0.028)。皮损组内细胞间的相互作用数量、强度和通路富集程度的活跃性均高于对照组,Treg细胞、CD8^(+)T细胞以及静息T细胞仅在银屑病组内与其他细胞有交互作用。拟时序分析显示免疫细胞主要分布在轨迹早期,单核细胞全程参与并在轨迹后期表达增加。除IL7R的表达量随着拟时序变化而减少外,其余hub基因的表达量均一直增加。预测转录因子NFKB1同时参与hub基因SPRR2A、IL1RN、IL7R和IER3的调控。结论SPRRs基因和IL7R的过度表达以及多种免疫细胞如Treg细胞、CD8^(+)T细胞、静息T细胞以及单核细胞的交互作用可能通过MAPK和NF-κB途径参与银屑病的关键致病过程。本研究结果可为进一步研究银屑病的致病机制奠定理论依据。Objective To investigate the potential pathogenic biomarkers associated with psoriasis and cellular heterogeneity at the level of single cells.Methods Single cell sequencing data from psoriatic skin biopsies were subjected to quality control,dimensional-reduction clustering and cell annotation.Cell clusters were analyzed for differentially expressed genes(DEGs),cell communication and pseudotime analysis.And TFs-miRNA-hub genes regulatory networks were constructed.Results 6 hub genes were identified by multialgorithm,SPRR2A,SPRR2D,IL7R,IL1RN,IER3,and LCN2.NF-κB,TNF and MAPK signaling pathway were shown to be significantly enriched.The expression of SPRRs was significantly upregulated in endothelial cells,fibroblasts and immune cells such as Langerhans cells,CD8^(+)T cells,M1 macrophages,resting T cells,while IL7R was predominantly expressed in immune cells.A dramatic increase of the proportion of fibroblasts,dendritic cells,Treg cells and CD8^(+)T cells in psoriatic skin lesions(LS)(P=0.023,P=0.007,P=0.046,P=0.028).The activeness of interactions number,interaction strength and level of pathway enrichment of cell communication were higher in LS than in the healthy control group.Fibroblasts,dendritic cells and resting T cells only interact with other cells in LS.Pseudotime analysis showed that immune cells were mainly distributed in early trajectory,and monocytes involved in the whole trajectory and increased in later trajectory.The expression of hub genes(except IL7R)were all increased along with pseudotime.Transcription factors,NFKB1,was involved in the regulation of hub genes(SPRR2A,IL1RN,IL7R and IER3)transcription.Conclusion Overexpression of SPRRs and IL7R,and interactive effects of immune cells may be involved in the pathogenic processes of psoriasis via the NF-κB and MAPK pathways,which lays the theoretical basis for further in-depth study of the pathogenic mechanism of psoriasis.

关 键 词：银屑病皮损 单细胞测序 富含脯氨酸小蛋白 免疫细胞 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]