METHODS: We used the RNA sequencing data from the GEO database for KIRC GSE121636 and normal kidney tissue GSE131685, and performed single-cell analysis for cluster identification, pathway enrichment, and CD8+ T cell-associated gene identification. Subsequently, the significance of different CD8+ T-cell associated gene subtypes was elucidated by consensus clustering, pathway analysis, mutated gene analysis, and KIRC immune microenvironment analysis in the TCGA-KIRC disease cohort. Single gene analysis identified LAG3 as the most critical CD8+ T-cell-associated gene and its function was verified by cell phenotype and immunohistochemistry in KIRC.
RESULTS: In the present study, CD8+ T-cell associated genes in KIRC were screened, including GZMK, CD27, CCL4L2, FXYD2, LAG3, RGS1, CST7, DUSP4, CD8A, and TRBV20-1 and an immunological risk prognostic model was constructed (risk score = - 0.291858656434841*GZMK - 0.192758342489394*FXYD2 + 0.625023643446193*LAG3 + 0.161324477181591*RGS1 - 0.380169045328895*DUSP4 - 0.107221347575037*TRBV20-1). LAG3 was identified and proved as the most critical CD8+ T cell-associated gene in KIRC.
CONCLUSIONS: We proposed and constructed an immunological risk prognostic model for CD8+ T cell-associated genes and identified LAG3 as a pivotal gene for KIRC progression and CD8+ T-cell infiltration. The model comprehensively explained the immune microenvironment and provided novel immune-related therapeutic targets and biomarkers in KIRC.
方法:我们将来自GEO数据库的RNA测序数据用于KIRCGSE121636和正常肾组织GSE131685,并进行单细胞分析以进行簇识别,途径富集,和CD8+T细胞相关基因鉴定。随后,通过共识聚类阐明了不同CD8+T细胞相关基因亚型的意义,途径分析,突变基因分析,TCGA-KIRC疾病队列中的KIRC免疫微环境分析。单基因分析确定LAG3是最关键的CD8+T细胞相关基因,其功能在KIRC中通过细胞表型和免疫组织化学得到证实。
结果:在本研究中,筛选KIRC中CD8+T细胞相关基因,包括GZMK,CD27,CCL4L2,FXYD2,LAG3,RGS1,CST7,DUSP4,CD8A,和TRBV20-1并建立了免疫风险预后模型(风险评分=-0.291858656434841*GZMK-0.192758342489394*FXYD2+0.625023643446193*LAG3+0.161324477181591*RGS1-0.380169045328895*DUSP4-0.1072213475V737501LAG3被鉴定并证明是KIRC中最关键的CD8+T细胞相关基因。
结论:我们提出并构建了CD8+T细胞相关基因的免疫风险预测模型,并确定LAG3是KIRC进展和CD8+T细胞浸润的关键基因。该模型全面解释了免疫微环境,并在KIRC中提供了新的免疫相关治疗靶标和生物标志物。