基于单细胞转录组测序探索患者来源类器官-胰腺癌原发性肿瘤来源免疫细胞的作用机制  

Exploring the acting mechanism of patient-derived organoids-pancreatic cancer primary tumor-derived immune cells based on single-cell RNA sequencing

作　　者：徐应翠 陆合明[2] 曹婷婷 XU Yingcui;LU Heming;CAO Tingting(Graduate School of Guangxi University of Chinese Medicine,Nanning 530000,China;Department of Radiotherapy,the People′s Hospital of Guangxi Zhuang Autonomous Region,Nanning 530021,China)

机构地区：[1]广西中医药大学研究生院,南宁530000 [2]广西壮族自治区人民医院放疗科,南宁530021

出　　处：《中国临床新医学》2024年第12期1398-1405,共8页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

基　　金：广西重点研发计划项目(编号:桂科AB17195005);广西医疗卫生适宜技术开发与推广应用项目(编号:S2017082)。

摘　　要：目的基于单细胞转录组测序探索患者来源类器官(PDO)-胰腺癌原发性肿瘤来源免疫细胞的作用机制。方法从基因表达综合数据库(GEO)获取胰腺癌单细胞转录组测序数据集,从癌症基因组图谱(TCGA)数据库下载185例胰腺癌患者的临床相关数据。经质控、降维、聚类和细胞注释后,分析免疫细胞在胰腺癌原发性肿瘤和PDO上的构成和分布差异,筛选免疫细胞的显著差异基因(DEGs),CellChat分析免疫细胞之间的分子相互作用,识别免疫细胞通讯模式。随后针对中性粒细胞DEGs进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。最后分析FFAR2、RASGEF1C基因表达水平对患者生存预后的影响。结果该研究成功识别了22个细胞群,并进一步将其分成11类细胞,其中包括中性粒细胞在内的4类免疫细胞。提取免疫细胞单细胞转录组测序数据得到30141个细胞,其中包括PDO模型来源的2282个细胞和原发性肿瘤来源的7319个细胞,在不同来源的样本中,免疫细胞构成和数量存在显著差异。通过CellChat软件分析免疫细胞之间的分子相互作用关系以及免疫细胞的通讯模式,识别出转化生长因子-β(TGF-β)信号通路和中性粒细胞在胰腺癌发展中占有重要地位。另外研究发现,中性粒细胞可以同时激活TGF-β等多种信号模式和途径。中性粒细胞的DEGs主要参与了免疫反应调节、激活免疫反应运输、激活先天免疫应答等生物学过程,并富集于原发性免疫缺陷、哮喘等信号通路。选择中性粒细胞DEGs中研究较少的基因FFAR2、RASGEF1C进行生存分析,结果提示FFAR2、RASGEF1C高表达有助于改善胰腺癌患者的生存预后。结论该研究确定了胰腺癌原发性肿瘤和PDO之间的免疫细胞差异性,为胰腺癌的类器官研究奠定了基础。Objective To explore the acting mechanism of patient-derived organoids(PDO)-pancreatic cancer primary tumor-derived immune cells based on single-cell ribonucleic acid(RNA)sequencing.Methods Single-cell RNA sequencing datasets of pancreatic cancer was obtained from the Genome Expression Omnibus(GEO)database,and the relevant clinical data of 185 cases of pancreatic cancer patients were downloaded from the Cancer Genome Atlas(TCGA)database.After quality control,dimensionality reduction,clustering and cellular annotation,the differences in the composition and distribution of immune cells on pancreatic cancer primary tumors and PDO were analyzed,and the differentially expressed genes(DEGs)of immune cells were screened.Molecular interactions between immune cells were analyzed using CellChat,and the immune cell communication patterns were recognized.Subsequently,Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed for neutrophils DEGs.Finally,the effects of expression levels of FFAR2 and RASGEF1C genes on survival prognosis of the patients were analyzed.Results In this study,22 cell populations were successfully identified and further categorized into 11 types of different cells,among which there were 4 types of immune cells,including neutrophils.The single-cell RNA sequencing data of immune cells were extracted to obtain 30141 cells,including 2282 cells from PDO model and 7319 cells from primary tumors.There were significant differences in the composition and number of immune cells in the samples from different sources.The molecular interactions between immune cells and the communication patterns of immune cells were analyzed through CellChat,and the transforming growth factor-beta(TGF-β)signaling pathway and neutrophils were identified as playing an important role in the development of pancreatic cancer.In addition,the study found that neutrophils could activate TGF-βand other signaling patterns and pathways at the same time.DEGs of neutroph

关 键 词：单细胞转录组测序 胰腺癌 原发性肿瘤 患者来源类器官 

分 类 号：R735.9[医药卫生—肿瘤]