Epilepsy-on-a-Chip System for Antiepileptic Drug Discovery.
Epilepsy-on-a-Chip System for Antiepileptic Drug Discovery.
复制标题
DOI：
10.1109/tbme.2018.2871415
复制
发表时间：
2019-05
期刊：
影响因子：
--
通讯作者：
Berdichevsky Y
中科院分区：
文献类型：
--
作者：
Liu J;Sternberg AR;Ghiasvand S;Berdichevsky Y
海马切片培养物在切片后几天自发地发展慢性癫痫，并用作创伤后癫痫的体外模型。在这里，我们描述了一种混合微流体-微电极阵列（μflow-MEA）技术，该技术将微流体灌注网络和电极集成到小型化装置中，用于基于海马切片培养的抗癫痫药物发现。进行场电位模拟以帮助优化电极设计以检测类寄生虫群体活性。癫痫芯片模型通过慢性电记录、神经元存活定量和抗惊厥试验进行验证。为了展示μflow-MEA在药物发现中的应用，我们利用两阶段筛选平台来识别抗癫痫药物的潜在靶点。在第I阶段，进行乳酸盐和乳酸脱氢酶生物标志物测定以鉴定潜在的候选药物。在第II阶段，候选化合物用基于μflow-MEA的慢性电测定法重新测试，以提供生物标志物结果的电生理学确认。我们筛选了12种受体酪氨酸激酶抑制剂，其中EGFR/ErbB-2抑制剂和cFMS抑制剂被确定为新型抗癫痫化合物。这种癫痫芯片系统提供了快速解剖癫痫发生中复杂信号通路的方法，为高通量抗癫痫药物的发现铺平了道路。
Hippocampal slice cultures spontaneously develop chronic epilepsy several days after slicing and are used as an in vitro model of post-traumatic epilepsy. Here, we describe a hybrid microfluidic-microelectrode array (μflow-MEA) technology that incorporates microfluidic perfusion network and electrodes into a miniaturized device for hippocampal slice culture based antiepileptic drug discovery. Field potential simulation was conducted to help optimize the electrode design to detect seizure-like population activity. Epilepsy-on-a-chip model was validated by chronic electrical recording, neuronal survival quantification, and anticonvulsant test. To demonstrate the application of μflow-MEA in drug discovery, we utilized a two-stage screening platform to identify potential targets for antiepileptic drugs. In Stage I, lactate and lactate dehydrogenase biomarker assays were performed to identify potential drug candidates. In Stage II, candidate compounds were retested with μflow-MEA based chronic electrical assay to provide electrophysiological confirmation of biomarker results. We screened 12 receptor tyrosine kinases inhibitors, and EGFR/ErbB-2 inhibitor and cFMS inhibitor were identified as novel antiepileptic compounds. This epilepsy-on-a-chip system provides the means for rapid dissection of complex signaling pathways in epileptogenesis, paving the way for high-throughput antiepileptic drug discovery.