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

描述（由申请人提供）：我是一名年轻的神经科学家，毕业于田纳西大学，并在华盛顿大学接受博士后培训。我在神经科学的原理方面获得了扎实的知识，并以游离细胞和脑切片电生理学为中心获得了出色的技术专长。我的长期职业目标是建立一个研究实验室，作为一个独立的研究者，专注于了解和开发神经系统疾病的治疗方法。此外，我希望教授和培训卫生专业人员和科学家。为了实现这些目标，我打算在3-5年的奖学金期间致力于扩展我的科学知识和提高研究技能。本提案的科学活动将包括参加研讨会、期刊俱乐部、国家和国际会议;编写出版手稿;培训学生和初级博士后研究员。研究活动将包括为我学习新技术，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，以及进行基因突变。我将使用这些技术深入研究生酮饮食在婴儿期严重肌阵挛性癫痫（SMEI，也称为Dravet综合征）中的治疗特性，使用华盛顿大学（美国领先的研究机构之一）的小鼠模型。SMEI是儿童癫痫的一种非常恶性的形式，最近与导致1型钠（Nav 1.1）通道减少的突变有关。这些突变存在于编码Nav1.1通道的基因Scnla中;它们阻止通道表达或降低功能。在我们的实验室中，通过引入阻止Scnla基因表达的突变来创建SMEI的小鼠模型。这些突变小鼠具有减少的Nav 1.1钠电流，并再现人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癫痫发作。生酮饮食是一种热量限制的方案，其提供约4：1的脂肪与碳水化合物和蛋白质的组合比率。这种饮食通常更有效地管理难以控制的癫痫发作，如SMEI。然而，其抗癫痫疗效的机制尚不清楚。本研究的具体目的如下：1）确定生酮饮食在小鼠SMEI中的抗惊厥和抗癫痫作用; 2）评估生酮饮食治疗作用背后的神经元功能和钠通道表达的变化; 3）研究生酮饮食对“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对神经元兴奋性和癫痫易感性的影响，促进对生酮饮食作用机制的理解。此外，这项研究将提供新的见解，可能会建议未来的药物或非药物治疗SMEI。

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.