Project 3 - Development and investigation of murine models of channelopathy-associated epilepsy

项目 3 - 通道病相关癫痫小鼠模型的开发和研究

基本信息

批准号：
10247560
负责人：
Jennifer A Kearney
金额：
$ 54.99万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10247560
关键词：
Action Potentials Acute Address Advanced Development Affect American Animals Anticonvulsants Brain Brain region Cell model Cells Clinical Collaborations Communities Data Detection Development Disease Electrophysiology (science)Epilepsy Equilibrium Etiology Evaluation Functional disorder Genes Genetic Goals Hippocampus (Brain)Impairment In Vitro Induced pluripotent stem cell derived neurons Interneurons Investigation Ion Channel Ion Channel Gating Modeling Mus Mutation Neocortex Neonatal Neurologic Neuronal Dysfunction Neurons Pathogenesis Patients Pattern Pharmaceutical Preparations Pharmacology Phenotype Population Potassium Precision therapeutics Property Regulatory Element Research Seizures Series Slice Sodium Standardization Syndrome System Testing Therapeutic Transgenic Mice Translations Variant base biophysical properties cell type clinically actionable early onset effectiveness evaluation epileptic encephalopathies genetic architecture in vivo in vivo Model in vivo evaluation infancy insight mouse model nervous system disorder precision medicine predictive modeling prevent response synergism voltage

项目摘要

PROJECT SUMMARY – PROJECT 3 Epilepsy is a common neurological disorder that affects over 3 million Americans and has a substantial genetic contribution to its etiology. Mutation of voltage-gated ion channel genes (‘Channelopathies’), particularly voltage-gated sodium (NaV) and potassium (KV) channel genes, have emerged as a major cause of early onset epileptic encephalopathies. These severe epilepsy syndromes are often difficult to treat with existing therapies and are associated with adverse neurodevelopmental sequelae, making them a high priority for better treatment approaches like precision medicine. Functional characterization of a small number of epilepsy- associated voltage-gated ion channel mutations in heterologous expression systems have demonstrated a range of dysfunction, but it is presently difficult to extrapolate these results to in vivo effects. A major goal of our Center is to determine how well in vitro cellular models predict neuronal dysfunction and pharmacological responses in an intact brain. To accomplish this goal, Project 3 will focus on a series of representative mouse models with NaV and KV channel variants that cause prototypical patterns of dysfunction. We hypothesize that differences in the relative contribution of specific channels to excitability in various cell types within neuronal networks determine the net effect on excitation-inhibition balance and influence pharmacological response. Mouse models provide the opportunity to evaluate the effect of channel variants at the whole animal, cellular and network levels, as well as to investigate pharmacological responses. In Aim 1, we will develop mouse models to investigate NaV and KV channel variants associated with early onset epileptic encephalopathy. Mouse lines will be evaluated for neurological phenotypes and pharmacological response in vivo. In Aim 2, we will determine the impact of NaV and KV channel variants on channel properties and intrinsic cell excitability in acutely dissociated neurons isolated from mouse models, and then determine the effectiveness of pharmacological agents at normalizing channel activity and/or cell excitability in these neurons. These results will be compared with similar recordings from heterologous expression systems (Project 1) and patient-specific iPSC-derived neurons (Project 2) to establish important correlations between in vitro and in vivo models. In Aim 3, we will determine the impact of NaV and KV channel variants on intrinsic properties of neurons and consequent effects on network activity in brain slices, and then determine the effectiveness of pharmacological agents at normalizing aberrant cellular and network excitability. Results from Project 3 will provide mechanistic insight into the effects of channel dysfunction in intact brains, and determine therapeutic strategies that normalize excitation-inhibition balance and prevent/reduce seizures. Synergy between this project and Projects 1 and 2 include cross-platform comparisons of the same channelopathy-associated epilepsy variants, which will facilitate translation of results into valuable information for implementation of precision medicine in this common neurological disorder.

项目摘要 - 项目3 癫痫是一种常见的神经系统疾病，影响300万美国人，并且具有实质性的遗传对其病因的贡献。电压门控离子通道基因的突变（“通道病”），特别是电压门控钠（NAV）和钾（KV）通道基因已成为早期发作的主要原因癫痫性脑病。这些严重的癫痫综合征通常很难治疗现有疗法并与不良神经发育后遗症有关，使其成为更好的优先级治疗方法如精密医学。少量癫痫的功能表征异源表达系统中相关的电压门控离子通道突变已证明功能障碍范围，但很难将这些结果推断为体内效应。一个主要目标我们的中心是确定体外细胞模型如何预测神经元功能障碍和药理完整大脑中的反应。为了实现这一目标，项目3将重点放在一系列代表性的鼠标上具有导致功能障碍的原型模式的NAV和KV通道变体的模型。我们假设这一点特定渠道对神经元内各种细胞类型的刺激性的相对贡献的差异网络确定净影响刺激抑制平衡并影响药物反应。鼠标模型提供了评估通道变体对整个动物的效果的机会和网络水平以及研究药物反应。在AIM 1中，我们将开发鼠标研究与早期发作癫痫性脑病相关的NAV和KV通道变体的模型。将在体内评估小鼠系的神经表型和药物反应。在AIM 2中，我们将确定NAV和KV通道变体对通道性质的影响以及在从小鼠模型中分离出的急性解离神经元，然后确定这些神经元中的通道活性和/或细胞兴奋性的药理剂。这些结果将与异源表达系统（项目1）和患者特异性的类似记录进行比较 IPSC衍生的神经元（项目2）建立体外和体内模型之间的重要相关性。在 AIM 3，我们将确定NAV和KV通道变体对神经元内固有特性的影响因此对大脑切片中网络活动的影响，然后确定药理的有效性使异常的细胞和网络令人兴奋的代理。项目3的结果将提供机理深入了解通道功能障碍在完整大脑中的影响，并确定治疗策略将兴奋的抑制平衡归一化，并防止/减少癫痫发作。这个项目与项目之间的协同作用 1和2包括对同一通道疗法相关癫痫变体的跨平台比较，将有助于将结果转化为价值信息，以实施精确医学常见的神经系统疾病。