Modeling genetic risk for epilepsy using IPSC and animal models.

项目摘要 本建议书描述了一个有指导的职业发展计划和研究建议，旨在 帮助我过渡到成为一名独立的临床科学家，专门从事遗传学研究。 癫痫的决定因素和癫痫发生的潜在分子机制。癫痫影响1%的 并导致严重的发病率。尽管越来越多的遗传病因学的发现， 癫痫易感性，这一信息还没有导致疾病修饰治疗。更好地了解 遗传变化导致的癫痫发生机制可能使我们更接近疾病的发展， 修改治疗。内在细胞兴奋性、神经元组织和突触功能的改变是 导致癫痫中网络兴奋性增加的潜在机制。15q11.2基因座缺失 在1.4%的遗传性癫痫中发现，使其成为迄今为止确定的最常见的易感性位点之一 并导致CYFIP1基因的单倍不足。CYFIP1基因调节活性依赖性 兴奋性突触的翻译以及正常神经发生所需的动态肌动蛋白重排 和突触发育。我假设癫痫易感性和神经精神障碍的增加， 15q11.2 CNV患者是由CYFIP1表达改变引起的。在本申请中，我提出了一个 研究计划，将评估CYFIP1介导的功能后果和潜在机制， 使用体外人iPSC和体内小鼠模型的组合来观察变化。我的导师团队包括 约翰霍普金斯大学的尼古拉斯·马拉加基斯、德怀特·伯格斯和卡尔·斯塔夫斯特罗姆以及 宾夕法尼亚大学.我为这笔资金确定了4个短期培训目标和2个长期目标 期我的短期目标是（1）开发体外人源化模型，以验证 癫痫和兴奋/抑制（E/I）不平衡在人类遗传背景的背景下;（2）发展 并操作动物模型以探测癫痫易感性和E/I失衡（3）接受以下训练： 先进的电生理技术，（4）提高我在统计学上合理的实验设计的技能 和分析我的长期目标是（1）发展独立的研究事业， 在神经发育障碍的更广泛的背景下癫痫的机制和（2）进一步发展一个 临床小生境在照顾病人的遗传决定癫痫。我的职业发展计划和 我的导师团队将帮助我实现这些目标。该研究计划将在体外建立互补的 和体内模型系统，以研究潜在的机制，筛选治疗干预措施， 开发一个可推广到癫痫其他遗传风险因素研究的平台。这些 研究将为我的R01提交提供基础，在R01中，我将检查基因型依赖性变化 在E/I平衡中筛选新的药物靶点并预测体内和体外的药物反应， 我接受的培训将有助于我在自己的实验室进行独立研究。

Project Abstract This proposal describes a mentored career development plan and research proposal that is designed to facilitate my transition to becoming an independent clinician scientist with specialization in the genetic determinants of epilepsy and underlying molecular mechanisms of epileptogenesis. Epilepsy affects 1% of the population and results in significant morbidity. Despite increasing discoveries of genetic etiologies that increase epilepsy susceptibility, this information has not led to disease modifying treatments. A better understanding of the mechanisms of epileptogenesis as a result of genetic changes may bring us closer to developing disease modifying treatments. Alterations in intrinsic cell excitability, neuronal organization and synaptic function are potential mechanisms leading to increased network excitability seen in epilepsy. Deletion of the 15q11.2 locus is found in 1.4% of genetic epilepsies, making it one of the most common susceptibility loci identified to date and it results in haplo-insufficiency of the CYFIP1 gene. The CYFIP1 gene regulates activity dependent translation at excitatory synapses as well as dynamic actin rearrangements required for normal neurogenesis and synaptic development. I hypothesize that increased seizure susceptibility and neuropsychiatric disorders in patients with 15q11.2 CNV results from alterations in CYFIP1 expression. In this application, I propose a research plan that will evaluate the functional consequences and underlying mechanisms of CYFIP1-mediated changes using a combination of in vitro human iPSC and in vivo mouse models. My mentorship team includes Nicholas Maragakis, Dwight Bergles, and Carl Stafstrom at Johns Hopkins University and Hongjun Song at the University of Pennsylvania. I have identified 4 short term training goals and 2 long term goals for this funding period. My short term goals are to (1) develop in vitro humanized models to validate molecular mechanisms of epilepsy and excitation / inhibition (E/I) imbalance in the context of human genetic backgrounds; (2) develop and manipulate animal models to probe epilepsy susceptibility and E/I imbalance (3) receive training in advanced electrophysiological techniques, and (4) improve my skills in statistically sound experimental design and analysis. My long term goals are to (1) develop an independent research career investigating the mechanisms of epilepsy in the wider context of neurodevelopmental disabilities and (2) further develop a clinical niche in the care of patients with genetically determined epilepsy. My career development plan and mentorship team will allow me to achieve these goals. The research plan will establish complementary in vitro and in vivo model systems to investigate underlying mechanisms, screen for therapeutic interventions and develop a platform that is generalizable to the investigation of other genetic risk factors for epilepsy. These studies will provide a foundation for my R01 submission in which I will examine genotype dependent changes in E/I balance to screen for new drug targets and predict drug response in vivo and in vitro and the mentorship and training that I receive will facilitate my transition to independent research in my own laboratory.