[Single-cell RNA sequencing deciphers transcriptional profiles of hepatocytes in mouse with hepatic alveolar echinococcosis].

• DOI: 10.16250/j.32.1374.2022275
• Publication Date: 2023-06-29
• Authors: Quan Yang; Jia Wang; Xian Wang; Qingqing Cai; Xin Ge; Weimin Wang; Xiuping Han
• Keywords: Echinococcus multilocularis; Hepatic stellate cell

To investigate the cell composition and transcriptional characteristics in microenvironments of hepatic tissues mice at late stage Echinococcus multilocularis infection a single-cell level.Peri-lesion paired distal specimens were collected from two BALB/c (6 to 8 weeks old) infected with E. for RNA sequencing. The Seurat package R software was employed quality control data, multi-sample integration correction batch effects, uniform manifold approximation projection (UMAP) algorithm used clustering. Cell types annotated using classical marker genes. Differentially expressed genes screened each type through differential gene expression analysis, biological roles cells predicted Gene Ontology (GO) Kyoto Encyclopedia Genes Genomes (KEGG) enrichment analyses.A total 43 710 peri-lesion multilocularis-infected analyzed, classified into 11 types, including neutrophils, T cells, macrophages, granulocyte-monocyte progenitor B plasma basophils, stellate endothelial hepatocytes, platelets. largest population immune microenvironment tissues, five CD4+ subsets, CD8+ subsets phosphoantigen-reactive γδT cells. proportions helper cytotoxic decreased proportion 2 (Th2) increased relative tissues. In addition, differentially Th2 associated negative regulation system, highly correlated activation system.Single-cell sequencing deciphers distribution multilocularis, may be formation immunosuppressive microenvironments.［摘要］目的 从单细胞水平探究多房棘球蚴感染小鼠晚期阶段肝脏组织微环境细胞组成及其转录谱特征。方法 收 集2只多房棘球蚴感染BALB/c小鼠 (6~8周龄) 肝脏病灶旁组织和配对远端肝组织进行单细胞转录组测序。利用R软 件Seurat包对获得的数据进行质量控制、多样本整合和批次效应校正, 应用统一流形逼近与投影 (uniform projection, UMAP) 算法进行细胞聚类, 根据经典标记基因注释细胞类型。通过差异基因表达分析筛选各 细胞类型的差异表达基因, 进行基因本体论 (Gene Ontology, GO) 和京都基因与基因组百科全书 (Kyoto Genomes, KEGG) 功能富集分析, 预测细胞生物学作用。结果 对来自多房棘球蚴感染小鼠肝脏病灶旁和 远端肝组织的43 710个细胞进行了分析, 归类为11种细胞类型：中性粒细胞、T细胞、巨噬细胞、粒细胞-单核细胞祖细 胞、B细胞、浆细胞、嗜碱性粒细胞、肝星状细胞、内皮细胞、肝细胞、血小板。T细胞是组织微环境中占比最高的免疫细 胞, 包括5种CD4+ T细胞、2种CD8+ T细胞和磷酸抗原反应γδ T细胞。与病灶远端肝组织相比, 病灶旁肝组织中的CD4+ 辅助性T细胞和CD4+ 细胞毒性T细胞比例降低、辅助性T细胞2 (Th2细胞) 比例明显增高。Th2细胞差异表达基因主要 与免疫系统负调控过程相关, CD4+细胞毒性T细胞高表达基因与免疫系统激活相关。结论 通过单细胞转录组测序揭 示了多房棘球蚴感染小鼠肝组织微环境中的细胞组成和分布差异, 病灶旁肝组织中Th2细胞升高可能与免疫抑制性微 环境形成有关。.