Xia Zhongbin, Cheng Ruoying, Liu Qi, Zu Yuxin, Liao Shilu
Health Management Medicine Department, The Second Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, China.
The Second Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, China.
Biomol Biomed. 2025 Mar 7;25(4):965-975. doi: 10.17305/bb.2024.10996.
Cell death has long been a focal point in life sciences research, and recently, scientists have discovered a novel form of cell death induced by copper, termed cuproptosis. This paper aimed to identify genes associated with cuproptosis in systemic lupus erythematosus (SLE) through machine learning, combined with single-cell RNA sequencing (scRNA-seq), to screen and validate related genes. The analytical results were then experimentally verified. Two published microarray gene expression datasets (GSE65391 and GSE61635) from SLE and control peripheral blood samples were downloaded from the GEO database. The GSE65391 dataset was used as the training group, while the GSE61635 dataset served as the validation group. Differentially expressed genes from GSE65391 identified 12 differential genes. Nine diagnostic genes, considered potential biomarkers, were selected using the least absolute shrinkage and selection operator and support vector machine recursive feature elimination analysis. The receiver operating characteristic (ROC) curves for both the training and validation groups were used to calculate the area under the curve to assess discriminatory properties. CIBERSORT was used to assess the relationship between these diagnostic genes and a reference set of infiltrating immune cells. scRNA-seq data (GSE162577) from SLE patients were also obtained from the GEO database and analyzed. Experimental validation of the most important SLE biomarkers was performed. Twelve significantly different cuproptosis-related genes were identified in the GSE65391 training set. Immune cell analysis revealed 12 immune cell types and identified nine signature genes, including PDHB, glutaminase (GLS), DLAT, LIAS, MTF1, DLST, DLD, LIPT1, and FDX1. In the GSE61635 validation set, seven genes were weakly expressed, and two genes were strongly expressed in the treatment group. According to the ROC curves, PDHB and GLS demonstrated significant diagnostic value. Additionally, correlation analysis was conducted on the nine characteristic genes in relation to immune infiltration. The distribution of key genes in immune cells was determined using scRNA-seq data. Finally, the mRNA expression of the nine diagnostic genes was validated using qPCR.
细胞死亡长期以来一直是生命科学研究的焦点,最近,科学家们发现了一种由铜诱导的新型细胞死亡形式,称为铜死亡。本文旨在通过机器学习结合单细胞RNA测序(scRNA-seq)来识别系统性红斑狼疮(SLE)中与铜死亡相关的基因,以筛选和验证相关基因。然后对分析结果进行实验验证。从GEO数据库下载了两个已发表的来自SLE和对照外周血样本的微阵列基因表达数据集(GSE65391和GSE61635)。GSE65391数据集用作训练组,而GSE61635数据集用作验证组。来自GSE65391的差异表达基因鉴定出12个差异基因。使用最小绝对收缩和选择算子以及支持向量机递归特征消除分析选择了9个诊断基因,这些基因被视为潜在的生物标志物。使用训练组和验证组的受试者工作特征(ROC)曲线来计算曲线下面积,以评估判别特性。使用CIBERSORT评估这些诊断基因与一组浸润免疫细胞参考之间的关系。还从GEO数据库获得并分析了SLE患者的scRNA-seq数据(GSE162577)。对最重要的SLE生物标志物进行了实验验证。在GSE65391训练集中鉴定出12个与铜死亡显著相关的基因。免疫细胞分析揭示了12种免疫细胞类型,并鉴定出9个特征基因,包括PDHB、谷氨酰胺酶(GLS)、DLAT、LIAS、MTF1、DLST、DLD、LIPT1和FDX。在GSE61635验证集中,7个基因表达较弱,2个基因在治疗组中表达较强。根据ROC曲线分析显示,PDHB和GLS具有显著的诊断价值。此外,对这9个特征基因进行了与免疫浸润相关的相关性分析。使用scRNA-seq数据确定关键基因在免疫细胞中的分布。最后,使用qPCR验证了这9个诊断基因的mRNA表达。