基于单细胞组学预测肝内胆管癌根治性切除患者预后的研究

Prediction of prognosis of patients with radical resection of intrahepatic cholangiocarcinoma based on single cell omics

目的:基于单细胞组学分析肝内胆管癌(ICC)根治性切除术后患者的生存获益。方法:本研究为回顾性病例系列研究。整合基因表达综合数据库中4个数据集的ICC单细胞测序结果,共获得46例患者根治性切除样本,探索ICC微环境的特征。同时分析附有生存数据的EMBI数据库共100例ICC组织芯片数据,通过ssGSEA算法计算芯片数据每个样本中各个上皮细胞亚群的浸润丰度,探索与患者较差预后相关的关键上皮细胞亚群,并研究该亚群关键标志(marker)基因的临床价值。通过单因素与多因素 Cox 比例风险模型筛选预后相关marker基因,使用目标基因构建 Cox比例风险模型并绘制列线图,使用Kaplan-Meier 生存分析验证评分与患者预后的关系,通过受试者工作特征(ROC)曲线、校准曲线及决策曲线(DCA)对模型预测效能进行评估。结果:上皮细胞亚群几乎仅在肿瘤组织中浸润。表达 MT2A的上皮细胞亚群与患者更差的预后相关且浸润丰度很高,以该亚群浸润为特征的患者被定义为抗氧化型。经单因素与多因素Cox比例风险模型筛选该亚群的marker基因,其中 FILPIL、 NAKBIA、 PEG10、 SERPINB5是患者的独立预后因素。用这4个基因表达量构建Cox比例风险模型,根据该模型所区分的高风险组患者的术后1、3、5年总体生存率均低于低风险组患者( P值均<0.05);所构建的列线图具有较好的区分度及有效性。ROC曲线显示,模型1、3、5年预测的曲线下面积分别为0.779、0.844、0.845;经校准曲线与DCA检验,对比临床指标,该模型具有较好的预测效能。 结论:MT2A阳性上皮细胞亚群与ICC患者根治性切除术后的生存获益相关,可将ICC组织分为抗氧化型与非抗氧化型,抗氧化型预示患者预后较差,可考虑通过术后辅助治疗等手段改善生存。

Objectives To analyze the survival benefit of intrahepatic cholangiocarcinoma(ICC)radical resection based on single cell omics.Methods This is a retrospective case-series study.ICC single-cell sequencing was integrated from four data sets in the Gene Expression Omnibus Database,with a total of 46 patients undergoing radical resection,to explore the characteristics of the microenvironment.Microarray data of 100 ICC cases was analyzed in the EMBI database with survival data.The infiltration abundance of each epithelial cell cluster was calculated in each microarray data sample using the ssGSEA algorithm.The key epithelial cell cluster associated with poor patient outcomes was explored.The clinical value of key marker genes in this subgroup was studied.Prognostic marker genes were selected using the univariate and multivariate Cox proportional hazards(CoxPH)model.The The CoxPH model was constructed by the target genes and a nomogram was drawn.Kaplan-Meier survival analysis was used to verify the relationship between score and prognosis of patients.The predictive power of the model was evaluated by receiver operating characteristic(ROC)curves,calibration curves,and decision curve analysis(DCA).Results Epithelial cell clusters infiltrated almost exclusively in tumor tissue.The MT2A+epithelial cell subset was associated with a poorer prognosis for patients with a high invasion abundance and patients characterized by infiltration of this group were defined as antioxidant.After screening marker genes in this cluster using a univariate and multivariate CoxPH model,the following genes were found to be independent prognostic factors:FILPIL,NFKBIA,PEG10,SERPINB5.The CoxPH model was constructed using the four gene expression levels,and the survival rate of patients in the high-risk group was significantly lower than those in the low-risk group(all P<0.05).The constructed nomogram had good discrimination and validity.The ROC curve showed that the predicted area under the curve was 0.779,0.844 and 0.845 at 1,3 and 5 years,respectively.Compared to clinical indicators,the model had better predictive power using the calibration curve and the DCA test.Conclusions The MT2A^(+) epithelial cell group may be associated with the prognosis of patients with ICC,and the concept of ICC tissue typing of antioxidant and non-antioxidant types is proposed.The type of antioxidant may predict the poor prognosis of the patients,and postoperative adjuvant therapy and other means could be considered to improve the survival of the patients.