Table_2_Circulating MicroRNAs From Plasma Small Extracellular Vesicles as Potential Diagnostic Biomarkers in Pediatric Epilepsy and Drug-Resistant Epilepsy.DOCX

Pediatric epilepsy is a neurological condition that causes repeated and unprovoked seizures and is more common in 1–5-year-old children. Drug resistance has been indicated as a key challenge in improving the clinical outcomes of patients with pediatric epilepsy. In the present study, we aimed to identify plasma small extracellular vesicles (sEVs) derived microRNAs (miRNAs) from the plasma samples of children for predicting the prognosis in patients with epilepsy and drug-resistant epilepsy. A total of 90 children clinically diagnosed with epilepsy [46 antiepileptic drug (AED)-responsive epilepsy and 44 drug-resistant epilepsy] and 37 healthy controls (HCs) were enrolled in this study. RNA sequencing was performed to identify plasma sEVs derived miRNAs isolated from the children’s plasma samples. Differentially expressed plasma sEVs derived miRNAs were identified using bioinformatics tools and were further validated by reverse transcription-polymerase chain reaction and receiver operator characteristic (ROC) curve analysis. In the present study, 6 miRNAs (hsa-miR-125b-5p, hsa-miR-150-3p, hsa-miR-199a-3p, hsa-miR-584-5p hsa-miR-199a-5p, and hsa-miR-342-5p) were selected for further validation. hsa-miR-584-5p, hsa-miR-342-5p, and hsa-miR-150-5p with area under curve (AUC) values of 0.846, 0.835, and 0.826, respectively, were identified as promising biomarkers of epilepsy. A logistic model combining three miRNAs (hsa-miR-584-5p, hsa-miR-342-5p, and hsa-miR-199a-3p) could achieve an AUC of 0.883 and a six miRNAs model (hsa-miR-342-5p, hsa-miR-584-5p, hsa-miR-150-5p, hsa-miR-125b-5p, hsa-miR-199a-3p, and hsa-miR-199a-5p) could attain an AUC of 0.888. The predicted probability of multiple miRNA panels was evaluated for differentiating between drug-resistant children and drug-responsive children. The AUC of a six-miRNA panel (hsa-miR-342-5p, hsa-miR-584-5p, hsa-miR-150-5p, hsa-miR-125b-5p, hsa-miR-199a-3p, and hsa-miR-199a-5p) reached 0.823. We identified and confirmed plasma sEVs derived miRNA biomarkers that could be considered as potential therapeutic targets for pediatric epilepsy and drug-resistant epilepsy.

小儿癫痫（Pediatric epilepsy）是一种以反复无诱因发作为特征的神经系统疾病，在1~5岁儿童中发病率更高。耐药性是改善小儿癫痫患者临床结局的关键挑战。本研究旨在从儿童血浆样本中筛选血浆源性细胞外小囊泡（small extracellular vesicles, sEVs）携带的微小核糖核酸（microRNAs, miRNAs），以预测癫痫及耐药性癫痫患者的预后。本研究共纳入90例经临床确诊的癫痫患儿[46例抗癫痫药物（antiepileptic drug, AED）应答型癫痫及44例耐药性癫痫]以及37例健康对照（healthy controls, HCs）。采用RNA测序技术对分离自儿童血浆样本的血浆sEVs源性miRNAs进行筛查；通过生物信息学工具鉴定差异表达的血浆sEVs源性miRNAs，并借助逆转录聚合酶链反应及受试者工作特征（receiver operator characteristic, ROC）曲线分析完成验证。本研究最终选取6个miRNAs（hsa-miR-125b-5p、hsa-miR-150-3p、hsa-miR-199a-3p、hsa-miR-584-5p、hsa-miR-199a-5p及hsa-miR-342-5p）开展后续验证。其中，hsa-miR-584-5p、hsa-miR-342-5p与hsa-miR-150-5p的曲线下面积（area under curve, AUC）分别为0.846、0.835及0.826，被鉴定为具有应用前景的癫痫生物标志物。联合3个miRNAs（hsa-miR-584-5p、hsa-miR-342-5p及hsa-miR-199a-3p）的逻辑回归模型AUC可达0.883；联合6个miRNAs（hsa-miR-342-5p、hsa-miR-584-5p、hsa-miR-150-5p、hsa-miR-125b-5p、hsa-miR-199a-3p及hsa-miR-199a-5p）的模型AUC可达0.888。本研究评估了多种miRNA标志物组合区分耐药性癫痫患儿与抗癫痫药物应答型癫痫患儿的诊断效能，其中上述6-miRNA组合的AUC达0.823。本研究鉴定并验证了血浆sEVs源性miRNA生物标志物，其可作为小儿癫痫及耐药性癫痫潜在的治疗靶点。