Two-step therapeutic screen for mitochondrial epilepsies

描述（由申请人提供）： 这项转化研究提案的长期目标是开发新型治疗药物，用于治疗与线粒体功能障碍相关的癫痫。该提案旨在解决一个重要的NINDS倡议，旨在开发筛选模型和确定毁灭性神经系统疾病的候选疗法。癫痫发作是在患有遗传性线粒体疾病的儿童中观察到的最常见特征。本实验室的工作表明线粒体功能障碍在癫痫引起的脑损伤以及癫痫易感性中的作用。基于这些研究，假设线粒体功能障碍是治疗干预的有吸引力的靶点。该提案的目标是开发和验证一个两步筛选模型，以确定优先改善线粒体功能障碍的治疗药物，因此将有利于灾难性的儿童线粒体癫痫。该模型是基于一种缺乏线粒体锰超氧化物歧化酶（MnSOD或Sod 2）的突变小鼠品系，这些小鼠在出生后经常发生自发性癫痫发作。第一个筛选（特定目标1）将利用大鼠脑线粒体来选择降低线粒体氧化应激的化合物。将在Sod 2-/-小鼠的第二体内模型中测试来自该筛选的选定化合物。第二次治疗筛选（特定目的2）的目的是在B6 D2 F1 Sod 2-/-小鼠中开发线粒体功能障碍的体内模型。模型开发将涉及视频监测、EEG记录、线粒体酶学、氧化应激和生存分析。该模型将验证一系列的亲脂性金属卟啉催化抗氧化剂，旨在通过血脑屏障。体外和体内方法可以一起顺序地用于鉴定候选治疗剂。该筛选程序将使我们能够参与NINDS合作计划，该计划旨在专门开发用于临床开发的候选疗法。

DESCRIPTION (provided by applicant): The long-term goal of this translational research proposal is to develop novel therapeutic agents for the treatment of epilepsies associated with mitochondrial dysfunction. This proposal is intended to address an important NINDS intiative aimed at developing screening models and identifying candidate therapeutics for devastating neurological disorders. Epileptic seizures are the most common feature observed in children with inherited mitochondrial diseases. Work in this laboratory suggests an emerging role of mitochondrial dysfunction in seizure-induced brain injury as well as seizure susceptibility. Based on these studies, it is hypothesized that mitochondrial dysfunction is an attractive target for therapeutic intervention. The goal of this proposal is to develop and validate a two-step screening model to identify therapeutic agents that preferentially ameliorate mitochondrial dysfunction and would therefore benefit catastrophic childhood mitochondrial epilepsies. The model is based on a strain of mutant mice lacking mitochondrial manganese superoxide dismutase (MnSOD or Sod2), that develop frequent spontaneous seizures in postnatal life. The first screen (Specific Aim 1) will utilize rat brain mitochondria to select compounds that decrease mitochondrial oxidative stress. Selected compounds from this screen will be tested in a second in vivo model of Sod2-/- mice. The goal of the second therapeutic screen (Specific Aim 2) is to develop an in vivo model of mitochondrial dysfunction in B6D2F1 Sod2-/- mice. Model development will involve video monitoring, EEG recordings, mitochondrial enzymology, oxidative stress and survival analysis. The model will be validated with a series of lipophilic metalloporpyrin catalytic antioxidants designed to cross the blood-brain barrier. Together, the in vitro and in vivo approaches can be utilized sequentially to identify candidate therapeutic agents. This screening procedure will allow us to participate the the NINDS cooperative program designed to specifically develop candidate therapies for clinical development.