Genetic Determinants of Epilepsy in Murine Systems

项目摘要 发育性和癫痫性脑病（DEE）是个别罕见，但集体大量 以使人衰弱的癫痫发作和持续的神经系统合并症为特征的神经发育障碍。 专门的外显子组测序工作已经在DEE基因发现方面取得了前所未有的成功，三分之一 或者更多的病例被带到遗传学诊所，导致明确的遗传学诊断。这一新的见解， 反过来，人们对精确或个性化医疗产生了更高的期望，为那些 否则就没有很多缓解疾病的选择，因为传统的药物治疗对 大多数症状在任何给定的DEE。原则上，基因治疗提供了治疗任何症状的可能性， 因为它是基于在原位替换或消除缺陷基因。在临床的障碍中， 本发明的应用包括递送的模式和时机，特别是对于神经发育疾病，以及 关注对不需要关注的细胞类型中基因表达的意外影响的副作用。 本更新提案的目的和设计是直接使用现成的可操作的 小鼠模型，以研究DEE发展和进展中的关键问题以及基因治疗的前景。 疗法首先，我们将应用明确设计的新方法来检测癫痫样活动和发育 小鼠幼仔中的里程碑，代表了研究不足但最适合建模儿童疾病的年龄。 然后，我们将使用小鼠遗传学工具来驱动不同神经元类型或发育年龄的基因表达 在这些模型中，以确定细胞病因学和疾病发展的关键窗口。最后， 我们将把基于RNA的基因治疗努力扩展到这些模型，测试有效减轻致病性 特征，同时监测全球基因表达的伴随变化。为了实现这些目标，我们 组建了一个由四个实验室组成的团队，每个实验室都提供互补的专业知识。通过这种协同作用， 通过共同努力，我们期望在理解疾病的基础方面取得重大进展， DEE模型，有可能在临床上推进新的治疗方法。

PROJECT SUMMARY Developmental and Epileptic Encephalopathies (DEE) are individually rare but collectively substantial neurodevelopmental disorders characterized by debilitating seizures and unremitting neurological comorbidities. Dedicated exome sequencing efforts have led to unprecedented success in DEE gene discovery, with one-third or more cases that are brought to the genetics clinic resulting in a clear genetic diagnosis. This new insight has, in turn, created high expectations for precision or personalized medicine to deliver new therapies to families who otherwise do not have many options for disease mitigation, since conventional drug therapy is ineffective for most symptoms in any given DEE. In principle, gene therapy offers potential for treating any symptom caused by a defective gene because it is based on replacing or eliminating it in situ. Among the barriers to clinical application include the mode and the timing of delivery, particularly for neurodevelopmental disease, and the concern about side effects from unintended impact on gene expression in cell types that do not require attention. The purpose and the design of this renewal proposal is to address these barriers directly using ready manipulable mouse models to examine key issues in the development and progression of DEE and the prospects for gene therapy. First, we will apply new methods explicitly designed to detect epileptiform activity and developmental milestones in mouse pups, representing an understudied but most appropriate age to model a childhood disease. We will then use mouse genetics tools to drive gene expression in different neuron types or developmental ages in these models, in order to determine the cellular etiology and critical window for disease development. Last, we will extend RNA-based gene therapy efforts to these models, testing for both effective mitigation of pathogenic features while monitoring accompanying changes in global gene expression. To accomplish these goals we have assembled a team of four laboratories each contributing complementary expertise. Through this synergistic, collaborative effort, we expect to make significant strides in understanding the basis of disease in three striking models of DEE with the potential to advance new treatments in the clinic.