单细胞转录组测序溃疡性结肠炎脾虚湿蕴证患者的免疫细胞分子特征  

Molecular characteristic of immune cells in ulcerative colitis patients with spleen deficiency and dampness syndrome based on single-cell RNA sequencing

作　　者：张诗静 黄卓健 陶屈源 侯江涛[2] 彭彬 罗泳欣 李燕舞[4] 李辉标[2] 钟嘉敏 陈新林[1] Zhang Shijing;Huang Zhuojian;Tao Quyuan;Hou Jiangtao;Peng Bin;Luo Yongxin;Li Yanwu;Li Huibiao;Zhong Jiamin;Chen Xinlin(School of Basic Medical Science,Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong Province,China;First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510400,Guangdong Province,China;First Clinical Medical College,Guangzhou University of Chinese Medicine,Guangzhou 510400,Guangdong Province,China;Science and Technology Innovation Center,Pi-Wei Institute,Guangzhou University of Chinese Medicine,Guangzhou 510400,Guangdong Province,China)

机构地区：[1]广州中医药大学基础医学院,广东省广州市510006 [2]广州中医药大学第一附属医院,广东省广州市510400 [3]广州中医药大学第一临床医学院,广东省广州市510400 [4]广州中医药大学科技创新中心,脾胃研究所,广东省广州市510400

出　　处：《中国组织工程研究》2023年第33期5357-5362,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(81774451),项目负责人:陈新林。

摘　　要：背景:免疫细胞介导的组织破坏对溃疡性结肠炎的发生和发展至关重要,但其机制尚未阐明,同时溃疡性结肠炎脾虚湿蕴证患者缺乏客观的生物标志物。目的:基于单细胞转录组测序技术探讨溃疡性结肠炎脾虚湿蕴证患者的免疫细胞分子特征,寻找溃疡性结肠炎脾虚湿蕴证患者的生物标志物,为其临床诊断及治疗提供参考依据。方法:临床获取2例溃疡性结肠炎患者和2例健康对照者的结肠活检组织,建库后进行单细胞转录组测序,并对单细胞转录组测序数据进行聚类和降维分析、细胞类型注释、差异表达分析和细胞通讯分析等。结果与结论:(1)单细胞转录组测序数据聚类鉴定出29个细胞亚群,组成9种细胞类型;(2)对比健康对照组,溃疡性结肠炎组CD4+细胞毒性T细胞、初始CD8+T细胞、生发中心、S100AhighB细胞、活化B细胞、NK_C2细胞和单核细胞的表达增高(P<0.05);溃疡性结肠炎患者的Tfh细胞、NKT细胞和NK细胞的表达减少(P<0.05);(3)差异表达基因的功能富集显示,溃疡性结肠炎患者的免疫调控增加,NK细胞、浆细胞和Th17细胞存在代谢重编程;(4)细胞通讯分析共鉴定了14种细胞之间的相互作用,以单核细胞和内皮细胞为主;(5)两组人群的免疫细胞分子特征存在较大差异,这些细胞亚群可能与溃疡性结肠炎的发生和发展有关,可能是溃疡性结肠炎脾虚湿蕴证患者的生物标志。BACKGROUND:Immune-mediated tissue destruction is essential for the development and deterioration of inflammation in ulcerative colitis,but their roles remain unclear.Simultaneously,ulcerative colitis patients with spleen deficiency and dampness syndrome lack objective biomarkers.OBJECTIVE:To investigate the molecular characteristics of immune cells in ulcerative colitis patients with spleen deficiency and dampness based on single-cell RNA sequencing so as to provide a basis for its clinical diagnosis and treatment.METHODS:Colon tissues of two ulcerative colitis patients and two healthy controls were obtained.Single-cell RNA sequencing was performed.The following analyses were carried out for the single-cell RNA sequencing data:cluster analysis,dimensionality reduction analysis,cell type annotation,differential expression analysis and cell communication analysis.RESULTS AND CONCLUSION:(1)A total of 29 cell sub-types were identified according to clustering analysis in single cell transcriptome,which consisted of 9 cell types.(2)Compared with healthy controls,the ulcerative colitis patients with spleen deficiency and dampness syndrome had higher expression levels of CD4+cytotoxic T cells,naive CD8+T cells,germinal center,activated B cells,S100AhighB cells,NK_C2 cells and monocytes(P<0.05).Compared with healthy controls,Tfh cells,NKT cells and NK cells decreased in patients with ulcerative colitis(P<0.05).(3)Functional enrichment of differentially expressed genes showed that there were hyperinflammatory activation in immune cells and higher metabolic reprogramming of NK cells,plasma cells and helper T cells for ulcerative colitis patients.(4)Cell communication analysis identified 14 interactions among different cell types,mainly from monocytes and endothelial cells.(5)There are significant differences in molecular characteristics of immune cells between ulcerative colitis patients and healthy controls.These cell subsets may be related to the occurrence and development of ulcerative colitis,which may be a biomarker fo

关 键 词：溃疡性结肠炎 单细胞转录组测序 免疫细胞 脾虚湿蕴 分子特征 生物标记 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R574