单细胞转录组在晚期肺腺癌患者个体化治疗中的应用

Application of single cell transcriptome in individual treatment of advanced lung adenocarcinoma

目的:分析晚期肺腺癌患者肿瘤细胞异质性,探索肿瘤细胞亚群转录组特性以寻找个体特异的治疗方案。方法:从人类肿瘤相关的基因表达汇编(GEO)数据库下载肺腺癌单细胞转录组测序数据芯片编号GSE123902,包含收集于2018年至2019年于纪念斯隆-凯特琳癌症中心手术切除的8例原发肺癌组织,3例脑转移样本,1例骨转移样本,1例肾上腺转移样本。过滤、鉴定并分离肿瘤细胞,对表达数据进行标准化、降维处理并根据表达模式对细胞进行聚类,利用基因差异表达分析,基因集变异分析(GSVA)探索不同肿瘤细胞亚群的表达特性。结果:共分离Ⅳ期肺腺癌原发灶肿瘤细胞211个,脑转移灶肿瘤细胞965个,骨转移灶肿瘤细胞659个,肾上腺转移灶肿瘤细胞14个。样本间比较涉及肿瘤血管生成、上皮间充质转化、侵袭转移相关的基因和基因集,例如骨转移组明显上调的波形蛋白(VIM,logFC=3.185,校正后的 P<0.01),差异有统计学意义;膜微囊蛋白-1(CAV-1,logFC=2.757,校正后的 P<0.01),差异有统计学意义。单样本分析中,脑转移和骨转移灶中均鉴定出耐药相关细胞亚群,存在多个耐药相关基因上调,包括脑转移细胞亚群中上调的载脂蛋白2(LCN2, logFC=1.822,校正后的 P<0.01),差异有统计学意义、骨转移细胞亚群中上调的趋化因子1(CXCL1,logFC=1.604,校正后的 P<0.01),差异有统计学意义。 结论:通过单细胞转录组分析肿瘤细胞异质性,可为晚期肺腺癌患者寻找个体化治疗方案提供线索。

Objective To explore transcriptome characteristics and heterogeneity of tumor cell subgroups for optimal individual treatment,this study analyze the heterogeneity of tumor cells in patients with advanced lung adenocarcinoma.Methods The lung adenocarcinoma single-cell transcriptome sequencing data chip number GSE123902 were downloaded from human tumor related gene expression omnibus(GEO)database.The samples were obtained from patients with lung adenocarcinoma(LUAD)undergoing resection surgery at Memorial Sloan Kettering Cancer Center(MSKCC),including eight primary tumor samples,three brain metastasis samples,one bone metastasis sample and one adrenal metastasis sample.Firstly,we infiltrated,annotated and isolated the tumor cells,and then normalized data,and performed dimensional reduction.Finally,the cells were clustered according to the expression pattern.The differential gene expression genes and gene set variant analysis(GSVA)were used to explore the expression characteristics of different tumor cell subsets.Results A total of 1849 tumor cells was isolated from stageⅣprimary tumor,adrenal,brain and bone metastases.The inter-patient heterogeneity was involved in tumor angiogenesis,epithelial mesenchymal transition(EMT),genes associated with tumor invasion and metastasis,such as Vimentin(VIM,logFC=3.185,Pcorrected<0.01)and Caveolin-1(CAV-1,logFC=2.757,Pcorrected<0.01),suggesting that the Vimentin and Caveolin-1 were significant increased in bone metastasis.Subgroups of tumor cells associated with chemo-resistance were identified both in brain and bone metastases,such as Lipocalin 2(LCN2,logFC=1.822,Pcorrected<0.01,C-X-C motif chemokine ligand 1(CXCL1,logFC=1.604,Pcorrected<0.01).Conclusion Single cell transcriptome analysis of tumor cell heterogeneity can provide clues to look for a new and accurate individual treatment.