Table2_Identification of RNA N6-methyladenosine regulation in epilepsy: Significance of the cell death mode, glycometabolism, and drug reactivity.XLSX

Epilepsy, a functional disease caused by abnormal discharge of neurons, has attracted the attention of neurologists due to its complex characteristics. N6-methyladenosine (m6A) is a reversible mRNA modification that plays essential role in various biological processes. Nevertheless, no previous study has systematically evaluated the role of m6A regulators in epilepsy. Here, using gene expression screening in the Gene Expression Omnibus GSE143272, we identified seven significant m6A regulator genes in epileptic and non-epileptic patients. The random forest (RF) model was applied to the screening, and seven m6A regulators (HNRNPC, WATP, RBM15, YTHDC1, YTHDC2, CBLL1, and RBMX) were selected as the candidate genes for predicting the risk of epilepsy. A nomogram model was then established based on the seven-candidate m6A regulators. Decision curve analysis preliminarily showed that patients with epilepsy could benefit from the nomogram model. The consensus clustering method was performed to divide patients with epilepsy into two m6A patterns (clusterA and clusterB) based on the selected significant m6A regulators. Principal component analysis algorithms were constructed to calculate the m6A score for each sample to quantify the m6A patterns. Patients in clusterB had higher m6A scores than those in clusterA. Furthermore, the patients in each cluster had unique immune cell components and different cell death patterns. Meanwhile, based on the M6A classification, a correlation between epilepsy and glucose metabolism was laterally verified. In conclusion, the m6A regulation pattern plays a vital role in the pathogenesis of epilepsy. The research on m6A regulatory factors will play a key role in guiding the immune-related treatment, drug selection, and identification of metabolism conditions and mechanisms of epilepsy in the future.

癫痫是一类由神经元异常放电引发的功能性疾病，因其复杂的病理特征而受到神经科医师的广泛关注。N6-甲基腺苷酸（N6-methyladenosine, m6A）是一种可逆性mRNA修饰，在各类生物过程中发挥核心作用。然而，此前尚无研究系统评估m6A调控因子在癫痫中的作用。本研究通过对基因表达综合数据库（Gene Expression Omnibus, GEO）中数据集GSE143272进行基因表达筛选，在癫痫患者与非癫痫患者中鉴定出7个具有显著差异的m6A调控基因。本研究采用随机森林（random forest, RF）模型进行筛选，最终选取HNRNPC、WATP、RBM15、YTHDC1、YTHDC2、CBLL1及RBMX这7个m6A调控因子作为预测癫痫发病风险的候选基因。随后基于这7个候选m6A调控因子构建了列线图模型。决策曲线分析初步证实，癫痫患者可从该列线图模型中获益。基于筛选得到的显著m6A调控因子，本研究采用一致性聚类法将癫痫患者划分为两种m6A模式（clusterA与clusterB）。通过构建主成分分析算法计算每个样本的m6A评分，以量化不同的m6A模式。clusterB组患者的m6A评分显著高于clusterA组。此外，两个聚类组的患者分别具有独特的免疫细胞组分与不同的细胞死亡模式。同时，基于m6A分类，本研究还验证了癫痫与糖代谢之间的相关性。综上，m6A调控模式在癫痫的发病机制中发挥关键作用。未来针对m6A调控因子的研究将为指导癫痫的免疫相关治疗、药物选择以及代谢状态与机制的鉴定提供重要依据。