Therapeutic effects of ketogenic diet in a mouse model of severe myoclinic epilep

简介（由申请人提供）：我是一名年轻的神经科学家，在田纳西大学接受研究生培训，在华盛顿大学接受博士后培训。我有扎实的神经科学原理知识和出色的以游离细胞和脑切片电生理学为中心的技术专长。我的长期职业目标是建立一个研究实验室，作为一名独立的研究者，专注于了解和开发神经系统疾病的治疗方法。此外，我还希望教授和培训卫生专业人员和科学家。为了达到这些目标，我打算用3-5年的资助时间来扩展我的科学知识和提高我的研究技能。这项建议的科学活动将包括参加讨论会、期刊俱乐部、国家和国际会议；准备出版稿件；培养学生和初级博士后。研究活动将包括学习新的技术，包括通过脑电图（EEG）记录活体动物的大脑活动，培养细胞和表达离子通道，以及基因突变。我将使用这些技术，深入研究生酮饮食对婴儿期严重肌阵挛性癫痫（SMEI，即Dravet综合征）的治疗特性，使用华盛顿大学（美国领先的研究机构之一）的小鼠模型。SMEI是一种非常恶性的儿童癫痫，最近与1型钠（Nav 1.1）通道减少的突变有关。这些突变存在于编码Nav 1.1通道的scla基因中；它们阻止通道表达或降低功能。我们在实验室通过引入阻止scla基因表达的突变，建立了小鼠SMEI模型。这些突变小鼠降低了Nav 1.1钠电流，重现了人类SMEI的主要临床特征，包括严重癫痫发作和共济失调。大多数抗癫痫药物不能很好地控制人类SMEI发作。生酮饮食是一种限制卡路里的饮食方式，它提供的脂肪与碳水化合物和蛋白质的比例约为4:1。这种饮食通常更有效地管理难以控制的癫痫发作，如那些在SMEI。然而，其抗癫痫作用的机制尚不清楚。本研究的具体目的如下：1)确定生酮饮食对小鼠SMEI的抗惊厥和抗癫痫作用；2)探讨生酮饮食对神经功能和钠通道表达的影响；3)探讨生酮饮食对“脑”电压门控钠通道功能的影响。这些研究将通过揭示生酮饮食对Nav通道表达和功能以及神经元兴奋性和癫痫易感性的影响，进一步了解生酮饮食的作用机制。此外，本研究将为未来SMEI的药物或非药物治疗提供新的见解。

DESCRIPTION (provided by applicant): I am a young neuroscientist with a graduate training from the University of Tennessee and postdoctoral training from the University of Washington. I have acquired solid knowledge in the principles of neuroscience and excellent technical expertise centered on dissociated cell and brain slice electrophysiology. My long term career goals are to establish a research lab, as an independent investigator, focused on understanding and developing cures for neurological disorders. In addition, I expect to teach and train health professionals and scientists. To attain these goals, I intend to dedicate the 3-5 year award period of this grant toward expanding my scientific knowledge and enhancing research skills. The scientific activities of this proposal will include participating in seminars, journal clubs, national and international meetings; preparing manuscripts for publication; and training students and junior post-doctoral fellows. The research activities will include learning novel techniques for me including recording brain activity in live animals by electroencephalography (EEG), culturing cells and expressing ion channels, and making gene mutations. I will use these techniques to intensively investigate the therapeutic properties of ketogenic diet in severe myoclonic epilepsy in infancy (SMEI, as known as Dravet syndrome) using a mouse model of the disorder at the University of Washington, one of the leading research institutions in the nation. SMEI is a very malignant form of childhood epilepsy that has been associated recently with mutations causing reduced type 1 sodium (Nav 1.1) channels. These mutations are found in Scnla, the gene encoding Nav 1.1 channels; they prevent channel expression or reduce function. A mouse model of SMEI was created in our lab by introducing mutations that prevent Scnla gene expression. These mutant mice have reduced Nav 1.1 sodium current and reproduce the main clinical features of human SMEI including severe seizures and ataxia. Human SMEI seizures are not well-controlled with most anti epileptic drugs. Ketogenic diet is a calorie restricted regimen that provides a ratio of fat to carbohydrate and protein combined of about 4:1. This diet is often more efficacious in managing difficult-to-control seizures such as those in SMEI. However, the mechanisms responsible for its antiepileptic efficacy are not understood. The research proposed has the following specific aims: 1) To determine the anticonvulsive and antiepileptic effects of ketogenic diet in mouse SMEI; 2) to evaluate changes in neuronal function and sodium channel expression underlying the therapeutic effects of ketogenic diet; 3) to investigate the effects of ketogenic diet on the function of 'brain' voltage-gated sodium channels. These studies will advance the understanding of the mechanisms of action of ketogenic diet by revealing its impacts on Nav channel expression and function as well as on neuron excitability and seizure susceptibility. Furthermore, this study will give new insights that may suggest future pharmacological or non-pharmacological therapies for SMEI.