Two-step therapeutic screen for mitochondrial epilepsies

描述(由申请人提供)： 这项转译研究提案的长期目标是开发治疗与线粒体功能障碍相关的癫痫的新型治疗剂。这项建议旨在解决一项重要的NINDS倡议，旨在开发筛查模型并确定毁灭性神经疾病的候选疗法。癫痫发作是遗传性线粒体疾病儿童最常见的特征。该实验室的工作表明，线粒体功能障碍在癫痫诱导的脑损伤以及癫痫易感性中的作用正在显现。基于这些研究，我们假设线粒体功能障碍是治疗干预的一个有吸引力的靶点。这项建议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剂，从而使灾难性的儿童线粒体癫痫受益。该模型是基于一种缺乏线粒体锰超氧化物歧化酶(MnSOD或Sod2)的突变小鼠，这些小鼠在出生后出现频繁的自发性癫痫发作。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。从这一筛选中选择的化合物将在第二个Sod2-/-小鼠体内模型中进行测试。第二个治疗性筛选(特定目标2)的目标是建立B6D2F1 Sod2-/-小鼠体内线粒体功能障碍的模型。模型开发将涉及视频监控、脑电记录、线粒体酶学、氧化应激和生存分析。该模型将通过一系列亲脂性金属卟啉催化抗氧化剂进行验证，这些抗氧化剂旨在穿越血脑屏障。总之，体外和体内的方法可以顺序地用于确定候选的治疗剂。这一筛选程序将使我们能够参与NINDS合作计划，该计划旨在专门为临床开发开发候选疗法。

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.