Using Zebrafish to Advance our Understanding and Treatment of Epilepsy

描述（由申请人提供）：传统的癫痫药物发现计划以抗惊厥作用为目标，几乎完全依赖于成年啮齿动物的诱导癫痫发作模型。然而，现在已经描述了许多模拟人类癫痫的许多特征的遗传模型。这些模型提供了重要的信息，但不容易适应药物发现计划。作为一种简单的脊椎动物物种，适合快速遗传操作和高通量药物筛选，我们提出了一种替代方法，使用具有自发复发性癫痫发作表型的突变斑马鱼（Danio rerio）（即，癫痫）作为一个平台，以确定新的治疗药物难治性癫痫。我们最近开始探索斑马鱼自发单基因突变的可能性-特别是那些模仿儿童常见的灾难性癫痫形式-导致癫痫表型。具有功能丧失钠通道（Nav1.1/SCN 1A）突变的斑马鱼突变体（例如，在患有婴儿严重肌阵挛性癫痫和Dravet综合征的儿童中鉴定的基因家族）最近被我们的实验室鉴定为具有与人类状况相似的表型的癫痫斑马鱼。使用大规模转录组分析，自动化行为跟踪，体内电生理学和药理学方法，我们描述了一种新的方法，以进一步了解和潜在的治疗与Nav1.1突变相关的衰弱性癫痫疾病。在尤里卡的提案中，我们将使用这些突变斑马鱼（i）鉴定DS/SMEI治疗的分子靶点，（ii）鉴定DS/SMEI治疗的候选药物。我们的研究结果有望建立一种基于斑马鱼的替代方法，用于高通量小分子药物发现，主要用于儿童单基因癫痫疾病。

DESCRIPTION (provided by applicant): Traditional drug discovery programs for epilepsy target anticonvulsant effects and rely, almost exclusively, on induced seizure models in adult rodents. However, numerous genetic models that mimic many features of human epilepsies have now been described. These models provide important information but are not easily adapted to drug discovery programs. As a simple vertebrate species amenable to rapid genetic manipulation and high-throughput drug screening, we propose an alternative approach using mutant zebrafish (Danio rerio) with spontaneous recurrent seizure phenotypes (i.e., epilepsy) as a platform to identify new treatments for medically refractory epilepsy. We recently began to explore the possibility that spontaneous single-gene mutations in zebrafish - especially those mimicking catastrophic forms of epilepsy often seen in children - result in epileptic phenotypes. Zebrafish mutants featuring a loss-of-function sodium channel (Nav1.1/SCN1A) mutation (e.g., a gene family identified in children with Severe Myoclonic Epilepsy of Infancy and Dravet syndrome) were recently identified by our laboratory as epileptic zebrafish with phenotypes similar to the human condition. Using large-scale transcriptome analysis, automated behavioral tracking, in vivo electrophysiology and pharmacological approaches we describe a novel approach to further our understanding and potential treatment of debilitating epilepsy disorders associated with Nav1.1 mutation. In this EUREKA proposal we will use these mutant zebrafish in our efforts to (i) identify molecular targets for therapeutic treatment of DS/SMEI and (ii) identify drug candidates for therapeutic treatment of DS/SMEI. Our results promise to establish an alternative, zebrafish-based, approach for high-throughput small-molecule drug discovery targeted to monogenic epilepsy disorders seen primarily in children.