■中风是一种高发病率的疾病,残疾,和死亡率。免疫因素在缺血性卒中(IS)的发生中起着至关重要的作用,但其确切机制尚不清楚。本研究旨在通过识别免疫相关的生物标志物和评估免疫细胞的浸润模式来确定可能的免疫机制。
■我们从GEO下载了IS患者的数据集,应用R语言发现差异表达基因,并使用GO阐明了它们的生物学功能,KEGG分析,和GSEA分析。然后使用两种机器学习算法(最小绝对收缩和选择算子(LASSO)和支持向量机递归特征消除(SVM-RFE))获得集线器基因,并通过CIBERSORT揭示免疫细胞浸润模式。使用Cytoscape构建基因-药物靶网络和mRNA-miRNA-lncRNA调控网络。最后,我们使用RT-qPCR验证了hub基因,并应用逻辑回归方法构建了用ROC曲线验证的诊断模型.
■我们筛选了188个差异表达基因,其功能分析富集到多个免疫相关途径。六个hub基因(ANTXR2,BAZ2B,C5AR1,PDK4,PPIH,和STK3)使用LASSO和SVM-RFE进行鉴定。ANTXR2,BAZ2B,C5AR1、PDK4和STK3与中性粒细胞和γδT细胞呈正相关,与滤泡辅助性T细胞和CD8呈负相关,而PPIH则呈完全相反的趋势。免疫浸润显示单核细胞活性增加,巨噬细胞M0,中性粒细胞,和肥大细胞,IS组滤泡辅助性T细胞和CD8的浸润减少。ceRNA网络由306个miRNA-mRNA相互作用对和285个miRNA-lncRNA相互作用对组成。RT-qPCR结果表明,BAZ2B的表达水平,IS患者C5AR1、PDK4和STK3显著增高。最后,我们建立了基于这四个基因的诊断模型.模型的AUC值在训练集中被验证为0.999,在验证集中被验证为0.940。
■我们的研究探索了免疫相关的基因表达模块,为进一步研究IS的免疫调节治疗提供了具体依据。
UNASSIGNED: Stroke is a disease with high morbidity, disability, and mortality. Immune factors play a crucial role in the occurrence of ischemic stroke (IS), but their exact mechanism is not clear. This study aims to identify possible immunological mechanisms by recognizing immune-related biomarkers and evaluating the infiltration pattern of immune cells.
UNASSIGNED: We downloaded datasets of IS patients from GEO, applied R language to discover differentially expressed genes, and elucidated their biological functions using GO, KEGG analysis, and GSEA analysis. The hub genes were then obtained using two machine learning algorithms (least absolute shrinkage and selection operator (LASSO) and support vector machine-recursive feature elimination (SVM-RFE)) and the immune cell infiltration pattern was revealed by CIBERSORT. Gene-drug target networks and mRNA-miRNA-lncRNA regulatory networks were constructed using Cytoscape. Finally, we used RT-qPCR to validate the hub genes and applied logistic regression methods to build diagnostic models validated with ROC curves.
UNASSIGNED: We screened 188 differentially expressed genes whose functional analysis was enriched to multiple immune-related pathways. Six hub genes (ANTXR2, BAZ2B, C5AR1, PDK4, PPIH, and STK3) were identified using LASSO and SVM-RFE. ANTXR2, BAZ2B, C5AR1, PDK4, and STK3 were positively correlated with neutrophils and gamma delta T cells, and negatively correlated with T follicular helper cells and CD8, while PPIH showed the exact opposite trend. Immune infiltration indicated increased activity of monocytes, macrophages M0, neutrophils, and mast cells, and decreased infiltration of T follicular helper cells and CD8 in the IS group. The ceRNA network consisted of 306 miRNA-mRNA interacting pairs and 285 miRNA-lncRNA interacting pairs. RT-qPCR results indicated that the expression levels of BAZ2B, C5AR1, PDK4, and STK3 were significantly increased in patients with IS. Finally, we developed a diagnostic model based on these four genes. The AUC value of the model was verified to be 0.999 in the training set and 0.940 in the validation set.
UNASSIGNED: Our research explored the immune-related gene expression modules and provided a specific basis for further study of immunomodulatory therapy of IS.