目的:利用生物信息学分析法挖掘胚胎反复种植失败(recurrent implantation failure，RIF)患者的相关关键基因，鉴定人子宫内膜中细胞亚群及细胞间通讯，探讨其对子宫内膜容受性的影响。方法:从高通量基因表达(Gene Expression Omnibus，GEO)数据库中下载GSE103465数据集、GSE183837数据集和GSE223672数据集为研究样本，以RIF患者及对照组女性的子宫内膜组织数据为研究对象，使用R语言及Perl软件将原始数据经过质量分析后利用limma包筛选差异基因，使用筛选的差异基因进行基因本体论(gene ontology，GO)及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genome，KEGG)分析，构建蛋白互作网络(Protein-Protein Interaction Networks，PPI网络)，筛选关键基因，利用R语言相关包进行单细胞注释与分类，使用筛选的差异基因构建细胞间通讯图谱。结果:筛选出关键基因RPF2、DDX27、PWP2、CDC5L、NOP2、DCAF13、CEBPZ、FTSJ1、GNL3L和NSUN2，鉴定RIF患者子宫内膜的主要细胞类型分别为上皮细胞、自然杀伤细胞、粒单核祖细胞、内皮细胞和平滑肌细胞，差异基因富集的GRN、SPP1、PTN、MIF、MK信号通路通过受体-配体对参与细胞间通讯。结论:RIF患者子宫内膜容受性受损与RNA的修饰与加工异常，出现表观遗传修饰错误相关，还与炎症反应及代谢下降密切关联，细胞间通讯显示RIF患者子宫内膜上皮细胞骨桥蛋白-整合素介导的黏附与侵袭能力下降，可能是导致胚胎种植失败的原因。
Integration of Gene Expression Microarrays and Single-Cell Transcriptomics to Identify Intercellular Communication in the Endometrium of Recurrent Implantation Failure Patients
Objective:To explore the key genes related to patients with recurrent implantation failure(RIF)using bioinformatics analysis,to identify cell subpopulations and cell-cell communications in human endometrium,and to discuss the impact on endometrial receptivity.Methods:The GSE103465 dataset,GSE183837 dataset and GSE223672 dataset were downloaded from the Gene Expression Omnibus(GEO)database as research samples.Endometrial tissue data from RIF patients and control females were utilized.R language and Perl scripts were employed to conduct the quality analysis on the raw data,and the limma package was utilized for differential gene selection.The selected differentially expressed genes were subjected to gene ontology(GO)and Kyoto encyclopedia of genes and genome(KEGG)analysis,Protein-Protein Interaction Networks(PPI)construction and core gene selection.Using R language-related packages,single-cell annotation and classification were performed,followed by the construction of an intercellular communication map.Results:The identified key genes included RPF2,DDX27,PWP2,CDC5L,NOP2,DCAF13,CEBPZ,FTSJ1,GNL3L,and NSUN2.The main cell types in the endometrium of patients with RIF were identified as epithelial cells,NK cells,granulocyte-monocyte progenitors,endothelial cells,and smooth muscle cells.The enriched pathways involving GRN,SPP1,PTN,MIF,MK signaling pathways participate in the intercellular communication through receptor-ligand pairs.Conclusions:The impaired endometrial receptivity in RIF patients is associated with aberrant RNA modification and processing and epigenetic modification errors,as well as inflammatory response and decreased metabolism.Intercellular communication reveals that endometrial epithelial cells in RIF patients have a decreased capacity for osteoblastin-integrin-mediated adhesion and invasion,which may be the cause of embryo implantation failure.
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