肺动脉高压与铜死亡相关标记基因和免疫特征的分析研究

Bioinformatics Analysis of Cuproptosis-Related Marker Genes and Immunological Characterization for Pulmonary Arterial Hypertension

目的:肺动脉高压(PAH)是一种严重危害人类健康的心肺疾病,组织中铜浓度的异常会导致PAH进展。本研究旨在利用生物信息学分析PAH中铜死亡相关性基因表达及其免疫学特征。方法:首先利用GEO数据集获取表达差异基因,再通过加权相关网络分析(WGCNA)检索与铜死亡相关的PAH模块,并将其与差异基因取交集,进行最小绝对值选择与收缩算子LASSO分析,然后采用受试者工作特征(ROC)曲线和外部数据集验证模型的诊断价值。最后,使用单样本基因集富集分析量化PAH表达谱中免疫细胞的浸润水平及与特征基因的关系。结果:通过对3999个PAH表达差异性基因进行KEGG分析,发现差异基因主要在神经活性配体-受体相互作用、mTOR信号通路和cAMP信号通路中富集。将差异基因与19个铜死亡相关基因取交集,共获得3个基因LIAS、LIPT1和NFE2L2。WGCNA表明,绿黄色模块与PAH的3个铜死亡相关差异基因显著相关,其与PAH差异基因相交后共获得490个候选基因,之后通过LASSO分析获得了NRBP2、ATG3、NT5M、MED23、DNAL4、TRAPPC9和NEB这7个关键基因。GSE113439验证了7个关键基因的表达水平和诊断价值。免疫浸润结果揭示,活化的CD4^(+)T细胞、嗜酸性粒细胞、未成熟树突状细胞、滤泡辅助性T细胞、自然杀伤细胞、CD56dim NK细胞和辅助T细胞1与关键基因存在显著性关联。结论:铜死亡相关的PAH共有7个关键基因(NRBP2、ATG3、NT5M、MED23、DNAL4、TRAPPC9和NEB),均与免疫浸润密切相关。

Objective:Pulmonary arterial hypertension(PAH)is a serious cardio-respiratory disease.The development of PAH may be exacerbated by abnormal copper levels.In this study,copper-associated marker genes and the immunological profile of PAH were identified using bioinformatics analysis.Methods:Differentially expressed genes(DEGs)were identified using the GEO datasets.To identify candidate genes,we searched for cuproptosis-related PAH modules using weighted correlation network analysis(WGCNA),which were intersected with DEGs,and screened using least absolute shrinkage and selection operator(LASSO)analysis.Then the diagnostic value was clarified by receiver operating characteristic(ROC)curves and the external dataset.Finally,we used single sample gene set enrichment analysis(ssGSEA)to quantify the relative infiltration level of immune cells in the expression profile and their relationship with hub genes.Results:KEGG analysis of 3999 DEGs indicated that DEGs were significantly enriched mainly in neuroactive ligand-receptor interaction,the mTOR signalling pathway,and the cAMP signalling pathway.Three genes,including LIAS,LIPT1,and NFE2L2,were obtained by intersecting DEGs with 19 cuproptosis-related genes.WGCNA demonstrated that the green-yellow module was the most significantly associated with three cuproptosis-related DEGs in PAH.A total of 490 genes were overlapped by module genes and DEGs.As a result,there were 7 hub genes including NRBP2,ATG3,NT5M,MED23,DNAL4,TRAPPC9,and NEB obtained by LASSO analysis.Then,GSE113439 was used to validate the expression level of the 7 candidate genes and their diagnostic predictive performance.The immune infiltration results revealed that the hub genes were significant associated with activated CD4^(+)T cells,eosinophils,immature dendritic cells,follicular helper T cells,natural killer cells,CD56dim NK cells and Th1 cells.Conclusion:There are seven key genes,including NRBP2,ATG3,NT5M,MED23,DNAL4,TRAPPC9 and NEB,in PAH associated with cuproptosis,which are closely related to immune infiltration.