Data_Sheet_8_N6-methyladenosine modulation classes and immune microenvironment regulation in ischemic stroke.XLSX

N6-methyladenosine (m6A) modifications play an important role in the differentiation and regulation of immune cells. However, research on m6A in ischemic stroke (IS) is still in its infancy, and their role of the immune microenvironment remains unknown. In this study, we systematically assessed the modification classes of m6A regulators in IS based on the GEO database (GSE16561 and GSE22255). We found that in IS patients, IGF2BP2, IGF2BP1, and YTHDF2 expression was significantly upregulated, and ELAVL1, LRPPRC, METTL3, ALKBH5, CBLL1, and METTL14 expression was significantly downregulated. Seven IS-related genes (ELAVL1, IGF2BP2, LRPPRC, YTHDF2, ALKBH5, METTL14, and YTHDC1) were finally screened by logistic and least absolute shrinkage and selection operator (LASSO) regressions, and the AUC of the riskScore was 0.942, which was a good classification. For immune infiltration, there were highly significant differences in memory B cells, CD8 T cells, monocytes, activated dendritic cells, and mast cells between IS and normal samples. The IS samples were grouped into three classes by consistent clustering, and 15 m6A genes were differentially expressed in the different classes. Multiple infiltrating immune cells, immune-associated genes, and HLA-associated genes differed significantly across m6A modification classes, indicating the diversity and complexity of m6A modifications in the immune microenvironment of IS. Finally, 487 genes associated with the m6A modification class were identified, and 227 potential drugs were found. Our findings demonstrated that m6A modification plays a crucial role in the immune regulation of IS.

N6-甲基腺嘌呤（N6-methyladenosine，m6A）修饰在免疫细胞的分化与调控过程中发挥重要作用。然而，针对缺血性脑卒中（ischemic stroke，IS）中m6A修饰的相关研究仍处于起步阶段，其在免疫微环境中的具体作用尚未明确。本研究基于基因表达综合数据库（Gene Expression Omnibus，简称GEO）的GSE16561与GSE22255数据集，系统分析了缺血性脑卒中样本中m6A调控因子的修饰谱。研究发现，缺血性脑卒中患者体内IGF2BP2、IGF2BP1及YTHDF2的表达水平显著上调，而ELAVL1、LRPPRC、METTL3、ALKBH5、CBLL1及METTL14的表达则显著下调。通过logistic回归与最小绝对收缩和选择算子（least absolute shrinkage and selection operator，LASSO）回归联合分析，最终筛选得到7个与缺血性脑卒中相关的核心基因（ELAVL1、IGF2BP2、LRPPRC、YTHDF2、ALKBH5、METTL14及YTHDC1），该风险评分模型的曲线下面积（Area Under the Curve，AUC）达0.942，展现出极佳的分类效能。在免疫浸润分析方面，缺血性脑卒中样本与正常对照样本在记忆B细胞、CD8阳性T细胞、单核细胞、活化树突状细胞及肥大细胞的浸润水平上存在极显著差异。通过一致性聚类将缺血性脑卒中样本划分为3个亚型，不同亚型间共有15个m6A基因呈现差异表达。不同m6A修饰亚型的浸润免疫细胞、免疫相关基因及人类白细胞抗原（Human Leukocyte Antigen，HLA）相关基因的表达均存在显著差异，这表明缺血性脑卒中免疫微环境中的m6A修饰具有高度的多样性与复杂性。本研究最终鉴定出487个与m6A修饰亚型相关的差异基因，并筛选得到227种潜在治疗药物。本研究结果证实，m6A修饰在缺血性脑卒中的免疫调控过程中发挥关键作用。