Temporal lobe epilepsy is the most common drug-resistant form of epilepsy in adults. The reorganization of neural networks and the gene expression landscape underlying pathophysiologic network behavior in brain structures such as the hippocampus has been suggested to be controlled, in part, by microRNAs. To systematically assess their significance, we sequenced Argonaute-loaded microRNAs to define functionally engaged microRNAs in the hippocampus of three different animal models in two species and at six time points between the initial precipitating insult through to the establishment of chronic epilepsy. We then selected commonly up-regulated microRNAs for a functional in vivo therapeutic screen using oligonucleotide inhibitors. Argonaute sequencing generated 1.44 billion small RNA reads of which up to 82% were microRNAs, with over 400 unique microRNAs detected per model. Approximately half of the detected microRNAs were dysregulated in each epilepsy model. We prioritized commonly up-regulated microRNAs that were fully conserved in humans and designed custom antisense oligonucleotides for these candidate targets. Antiseizure phenotypes were observed upon knockdown of miR-10a-5p, miR-21a-5p, and miR-142a-5p and electrophysiological analyses indicated broad safety of this approach. Combined inhibition of these three microRNAs reduced spontaneous seizures in epileptic mice. Proteomic data, RNA sequencing, and pathway analysis on predicted and validated targets of these microRNAs implicated derepressed TGF-β signaling as a shared seizure-modifying mechanism. Correspondingly, inhibition of TGF-β signaling occluded the antiseizure effects of the antagomirs. Together, these results identify shared, dysregulated, and functionally active microRNAs during the pathogenesis of epilepsy which represent therapeutic antiseizure targets.

颞叶癫痫是成人中最常见的耐药性癫痫形式。有人建议，神经网络的重组和基因表达在脑结构（如海马体）中的病理生理网络行为的基础，部分地受microRNA的控制。为了系统地评估它们的重要性，我们对加载了Argonaute的microRNA进行了测序，以定义两种物种中三种不同动物模型的海马中功能性参与的microRNA，并且在从最初的发作到慢性癫痫发作之间的六个时间点进行了定义。然后，我们使用寡核苷酸抑制剂为功能性体内治疗筛选选择了通常上调的microRNA。Argonaute测序产生了14.4亿个小RNA读数，其中多达82％是microRNA，每个模型检测到400多个独特的microRNA。在每个癫痫模型中，大约一半的检测到的microRNA失调。我们优先考虑了在人类中完全保守的通常上调的microRNA，并针对这些候选靶标设计了定制的反义寡核苷酸。敲低miR-10a-5p，miR-21a-5p和miR-142a-5p时观察到抗癫痫表型，电生理分析表明该方法具有广泛的安全性。联合抑制这三种microRNA可减少癫痫小鼠的自发性癫痫发作。这些微RNA的预测和验证目标的蛋白质组学数据，RNA测序和途径分析表明，解除抑制的TGF-β信号传导是一种共有的癫痫发作改变机制。相应地，TGF-β信号转导的抑制阻断了拮抗剂的抗癫痫发作作用。一起，