Modeling genetic risk for epilepsy using IPSC and animal models.

使用 IPSC 和动物模型对癫痫遗传风险进行建模。

• 批准号: 10197233
• 负责人: Christa Habela
• 金额: $ 19.93万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10197233
• 关键词: Actins; Acute; Affect; Angelman Syndrome; Animal Model; Biological Models; Cell Line; Cells; Clinical; Control Animal; Data; Development; Development Plans; Disease; Electrophysiology (science); Epilepsy; Epileptogenesis; Equilibrium; Excitatory Synapse; Experimental Animal Model; Experimental Designs; FMRP; Foundations; Functional disorder; Funding; Future; Gene Deletion; Generalized Epilepsy; Genes; Genetic; Genetic Determinism; Genetic Models; Genetic Predisposition to Disease; Genetic Risk; Genotype; Glutamates; Goals; Human; Human Genetics; In Vitro; Induced pluripotent stem cell derived neurons; Inhibitory Synapse; Intellectual functioning disability; Interneurons; Investigation; Knock-out; Laboratories; Lead; Measures; Mediating; Mentors; Mentorship; Modeling; Molecular; Morbidity - disease rate; Morphology; Mus; Mutation; Neurodevelopmental Disability; Neurodevelopmental Disorder; Neurologic; Neurons; Neurotransmitters; Other Genetics; Parvalbumins; Pathologic; Patient Care; Patients; Pennsylvania; Pharmacology; Phenotype; Physiology; Population; Positioning Attribute; Predisposition; Property; Proteins; Regulation; Research; Research Proposals; Rhodopsin; Risk; Risk Factors; Rodent; Rodent Model; Role; Schizophrenia; Scientist; Seizures; Signal Transduction; Slice; Structure; Susceptibility Gene; Synapses; Techniques; Therapeutic Intervention; Training; Translations; Universities; career; career development; design; drug response prediction; excitatory neuron; expectation; experimental analysis; experimental study; gamma-Aminobutyric Acid; genetic risk factor; glutamatergic signaling; hippocampal pyramidal neuron; immunocytochemistry; improved; in vivo; in vivo Model; induced pluripotent stem cell; inhibitory neuron; insight; mouse model; multi-electrode arrays; nerve stem cell; neural network; neurodevelopment; neurogenesis; neuron development; neuropsychiatric disorder; new therapeutic target; novel; research and development; response; risk variant; skills; sound; synaptic function; synaptogenesis

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.

项目摘要 该提案描述了一个受指导的职业发展计划和研究提案，旨在 帮助我过渡成为一名独立的临床科学家，专门从事遗传研究 癫痫的决定因素和癫痫发生的潜在分子机制。癫痫影响 1% 人口并导致显着的发病率。尽管越来越多的遗传病因学发现增加了 癫痫易感性，这一信息并未导致疾病治疗。更好地理解 遗传变化导致的癫痫发生机制可能会让我们更接近患上疾病 修改治疗方法。内在细胞兴奋性、神经元组织和突触功能的改变 导致癫痫中网络兴奋性增加的潜在机制。 15q11.2 基因座的删除 存在于 1.4% 的遗传性癫痫中，使其成为迄今为止发现的最常见的易感位点之一 它导致 CYFIP1 基因的单倍体不足。 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。我假设癫痫发作易感性和神经精神疾病增加 15q11.2 CNV 患者的 CYFIP1 表达变化所致。在这个应用程序中，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 使用体外人类 iPSC 和体内小鼠模型的组合进行改变。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 (Nicholas Maragakis)、德怀特·伯格斯 (Dwight Bergles) 和卡尔·斯塔夫斯特罗姆 (Carl Stafstrom) 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 时期。我的短期目标是（1）开发体外人源化模型来验证分子机制 人类遗传背景下的癫痫和兴奋/抑制（E/I）失衡； (2) 开发 并操纵动物模型来探究癫痫易感性和 E/I 失衡 (3) 接受以下方面的培训 先进的电生理技术，以及（4）提高我在统计上合理的实验设计方面的技能 和分析。我的长期目标是（1）发展独立的研究生涯，调查 在更广泛的神经发育障碍背景下的癫痫机制，以及（2）进一步开发 治疗遗传性癫痫患者的临床利基。我的职业发展计划和 导师团队将使我实现这些目标。该研究计划将建立互补的体外 和体内模型系统，以研究潜在机制、筛选治疗干预措施和 开发一个可推广到癫痫其他遗传风险因素调查的平台。这些 研究将为我提交的 R01 奠定基础，我将在其中检查基因型依赖性变化 在 E/I 平衡中筛选新药物靶点并预测体内和体外药物反应以及指导 我接受的培训将有助于我过渡到在自己的实验室进行独立研究。