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Mechanisms and Models of SPTAN1 Epileptic Encephalopathy

SPTAN1癫痫性脑病的机制和模型

基本信息

批准号：
10053733
负责人：
Yu Wang
金额：
$ 19.01万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-12-01 至 2021-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10053733
关键词：
Actins Acute Adhesions Animals Axon Binding Brain C-terminal Cell Communication Cell Culture Techniques Cell model Cells Clustered Regularly Interspaced Short Palindromic Repeats Complex Cytoskeleton DNA DNA Sequence Alteration Data Development Development Plans Developmental Delay Disorders Disease Disease model Disinhibition Doctor of Medicine Doctor of Philosophy Dominant-Negative Mutation Dyskinetic syndrome Electrophysiology (science)Electroporation Elements Embryo Epilepsy Epileptogenesis Excitatory Synapse Fibroblasts Foundations Functional disorder Funding Future Genes Genetic Genetic study Goals Grant Heterodimerization Human Impaired cognition In Vitro Infant Infantile spasms Inhibitory Synapse Intellectual functioning disability Knowledge Laboratories Lead Light Link Manuscripts Mechanical Stress Mediating Medicine Membrane Methods Modeling Molecular Mus Mutate Mutation Nature Neurologist Neurology Neurons Neurosciences Paper Pathogenicity Patients Periodicity Physicians Play Process Prosencephalon Proteins Publishing Rattus Reporter Research Role Scientist Seizures Side Slice Spectrin Structure Study models System Testing Therapeutic Studies Training Training Activity Viral Viral Vector Work Writing base beta Spectrin career career development childhood epilepsy de novo mutation drug use screening early childhood epileptic encephalopathies excitatory neuron genetic manipulation genome editing hippocampal pyramidal neuron human model in utero in vitro Model in vivo in vivo Model induced pluripotent stem cell infancy inhibitory neuron innovation insight migration mutant nervous system disorder neurodevelopment neuron development novel overexpression postsynaptic skills stem cell biology stem cell model success synaptogenesis tool

项目摘要

Abstract/Project Summary Applicant: Dr. Wang holds M.D. and Ph.D. degrees, and has completed specialized training in both neurology and epilepsy. During his Ph.D. work, he published 9 manuscripts including three first-author papers in Neuroscience and one second-author paper in Nature Medicine. During his neurology training, he conducted a genetic study on familial paroxysmal kinesigenic dyskinesia and co-authored one paper (under review). Research Plan: Defining how genetic mutations that cause infantile epileptic encephalopthies (IEEs) disrupt neurodevelopment should provide conceptual insights broadly relevant to understanding brain development and epilepsy mechanisms. Using CRISPR genome editing methods, in utero electroporation (IUE) and human induced pluripotent stem cell (iPSC) cultures, I propose to study the function of SPTAN1, a recently identified gene (encoding alpha II-spectrin) mutated in patients presenting with IEE. My preliminary data show that SPTAN1 regulates critical aspects of neuronal development, including axonal initial segment (AIS) formation, process outgrowth and clustering of postsynaptic proteins. My central hypotheses are that SPTAN1 plays critical roles in neurodevelopment through regulating neuronal migration, process outgrowth and synapse formation, and that disinhibition of excitatory neurons leads to seizures in SPTAN1 IEE. I propose two specific aims to test my hypotheses: 1) To determine how deleting Sptan1 or overexpressing mutant SPTAN1 in cortical pyramidal neurons alters brain development and network excitability; and 2) To establish how mutations in SPTAN1 influence excitatory and inhibitory cortical neuron development and excitability using patient-derived iPSCs. This innovative proposal combines a rat IUE in vivo model (Aim 1) and an in vitro model with patient-derived neurons (Aim 2). These models will be invaluable tools to study brain development, and will also serve as an entry point for future drug screening using patient-derived neurons. Immediate and long-term career goals: My career goal is to become an independent physician-scientist focusing on brain development and genetic epilepsies. My long-term goal is to understand how genetic mutations alter neurodevelopment and ultimately lead to seizures and intellectual disability. My short-term goal is to obtain knowledge (e.g., stem cell biology and electrophysiology) and skills (e.g., iPSC cultures, viral- based gene manipulation) in neuroscience to fill the gaps of my previous training in order to augment my chances of success as an independently funded neuroscientist. The key elements of this research career development plan are 5 key training goals. They are stem cell biology, electrophysiology principles, viral vector based genetic manipulation, grant writing and laboratory management skills.

摘要/项目摘要申请人：王博士拥有医学博士学位。和博士学位，并完成了神经病学和神经病学的专业培训，和癫痫。在他的PhD他发表了9篇手稿，其中包括三篇第一作者论文，神经科学和一个第二作者在自然医学论文。在他的神经病学培训期间，他进行了一次家族性阵发性运动诱发性运动障碍的遗传学研究，并与人合著了一篇论文（正在审查中）。研究计划：确定导致婴儿癫痫性脑病（IEE）的基因突变如何破坏神经发育应该提供与理解大脑发育广泛相关的概念性见解和癫痫机制。使用CRISPR基因组编辑方法，在子宫内电穿孔（IUE）和人诱导多能干细胞（iPSC）培养，我建议研究SPTAN 1的功能，SPTAN 1是最近发现的基因（编码α II-血影蛋白）在IEE患者中发生突变。初步数据显示 SPTAN 1调节神经元发育的关键方面，包括轴突起始段（AIS）形成，突触后蛋白质的生长和聚集。我的主要假设是SPTAN 1扮演着通过调节神经元迁移、突起生长和突触在神经发育中起关键作用在SPTAN 1 IEE中，兴奋性神经元的去抑制导致癫痫发作。我建议两个具体的目的是验证我的假设：1）为了确定缺失Sptan 1或过表达突变体SPTAN 1在皮质锥体神经元改变大脑发育和网络兴奋性;和2）为了建立如何 SPTAN 1突变影响兴奋性和抑制性皮质神经元发育和兴奋性使用患者来源的iPSC。这种创新的建议结合了大鼠IUE体内模型（Aim 1）和在体外的IUE模型（Aim 1）。体外模型与患者来源的神经元（目的2）。这些模型将是研究大脑的宝贵工具开发，也将作为未来使用患者来源的神经元进行药物筛选的切入点。近期和长期的职业目标：我的职业目标是成为一名独立的医生-科学家专注于大脑发育和遗传性癫痫我的长期目标是了解基因突变会改变神经发育，最终导致癫痫发作和智力残疾。我的短期目标是为了获得知识（例如，干细胞生物学和电生理学）和技能（例如，iPSC培养物，病毒- 基于基因操作）在神经科学中，以填补我以前的训练空白，以增加我的作为一个独立资助的神经科学家，成功的机会。这个研究生涯的关键要素发展计划是5个主要的培训目标。它们是干细胞生物学、电生理学原理、病毒载体基于遗传操作，赠款写作和实验室管理技能。