Multi-scale disease modeling of SCN2A-related epilepsy due to gain-of-function variants

由于功能获得性变异导致 SCN2A 相关癫痫的多尺度疾病模型

基本信息

批准号：
10525781
负责人：
SCOTT K. ADNEY
金额：
$ 15.75万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10525781
关键词：
Affect Animal Model Animals Area Award Benchmarking Biological Markers Biophysical Process Brain Cell Line Childhood Data Development Disease Disease model Electrodes Electroencephalography Electrophysiology (science)Epilepsy Equilibrium Exhibits Fostering Foundations Functional disorder Funding Future Genes Genetic Genetic Diseases Genetic Models Glutamates Hippocampus (Brain)Human Impairment Implant Induced pluripotent stem cell derived neurons Interneurons Intractable Epilepsy Ion Channel K-Series Research Career Programs Lead Link Mentors Missense Mutation Modeling Mus National Institute of Neurological Disorders and Stroke Neurodevelopmental Disorder Neurons Optics Output Pathogenicity Patients Pharmaceutical Preparations Phenotype Physicians Population Population Analysis Quality of life Recurrence Reporter Research Scientist Seizures Severities Signal Transduction Slice Sodium Channel Sodium Channel Blockers Sudden Death Synapses Teacher Professional Development Training United States Variant animal tissue autism spectrum disorder base career career development causal variant childhood epilepsy clinical phenotype collaborative environment design dravet syndrome early onset epileptic encephalopathies excitatory neuron gain of function genetic analysis genetic variant genome editing human stem cells human tissue in silico in vivo induced pluripotent stem cell infancy inhibitory neuron insight interest loss of function mortality mouse model multi-scale modeling network dysfunction neural circuit neurodevelopment neuronal circuitry neuronal excitability neurophysiology novel strategies programs research study spatiotemporal voltage

Affect Animal Model Animals Area Award Benchmarking Biological Markers Biophysical Process Brain Cell Line Childhood Data Development Disease Disease model Electrodes Electroencephalography Electrophysiology (science)Epilepsy Equilibrium Exhibits Fostering Foundations Functional disorder Funding Future Genes Genetic Genetic Diseases Genetic Models Glutamates Hippocampus (Brain)Human Impairment Implant Induced pluripotent stem cell derived neurons Interneurons Intractable Epilepsy Ion Channel K-Series Research Career Programs Lead Link Mentors Missense Mutation Modeling Mus National Institute of Neurological Disorders and Stroke Neurodevelopmental Disorder Neurons Optics Output Pathogenicity Patients Pharmaceutical Preparations Phenotype Physicians Population Population Analysis Quality of life Recurrence Reporter Research Scientist Seizures Severities Signal Transduction Slice Sodium Channel Sodium Channel Blockers Sudden Death Synapses Teacher Professional Development Training United States Variant animal tissue autism spectrum disorder base career career development causal variant childhood epilepsy clinical phenotype collaborative environment design dravet syndrome early onset epileptic encephalopathies excitatory neuron gain of function genetic analysis genetic variant genome editing human stem cells human tissue in silico in vivo induced pluripotent stem cell infancy inhibitory neuron insight interest loss of function mortality mouse model multi-scale modeling network dysfunction neural circuit neurodevelopment neuronal circuitry neuronal excitability neurophysiology novel strategies programs research study spatiotemporal voltage

展开

项目摘要

Project Summary Epilepsy affects up to 1% of the population worldwide, and 3 million in the United States alone. A growing proportion of pediatric epilepsies are tied to causative variants in ion channel genes, including the voltage-gated sodium channel gene SCN2A. The 2020 Epilepsy Research Benchmarks of NINDS prioritize identifying how genetic variants cause epilepsy and related neurodevelopmental disorders. SCN2A variants that manifest with loss-of-function are associated with severe neurodevelopmental disorders and late-onset epilepsy. On the other hand, gain-of-function SCN2A variants predominantly have a phenotype of early-onset epilepsy. The encoded sodium channel (NaV1.2) is highly expressed in excitatory glutamatergic neurons early in development, presenting a unique opportunity to examine how excitatory neuron dysfunction leads to early-onset epilepsy. Animal and human tissue-derived neuron models have brought mechanistic insight to how Dravet syndrome results in interneuron dysfunction and epilepsy. Among SCN2A-related diseases, animal models illuminate how loss-of-function leads to autism spectrum disorder with late-onset epilepsy. Due to lack of readily available disease models, there is sparse mechanistic understanding of how excitatory neuron dysfunction early in development leads to early-onset epilepsy. This proposal will exploit two early-onset epilepsy variants of SCN2A that have a convergent clinical phenotype yet divergent biophysical mechanisms. Patient-derived neuron models and mouse models provide the opportunity to define the point of mechanistic convergence at multiple scales: from single neurons to neural circuits influencing epilepsy phenotype. Aim 1 will determine how two gain- of-function SCN2A variants, encoding missense mutations M1879T and E430A, confer increased excitability by distinct mechanisms. Functional analysis of iPSC-derived neurons in isolation and in elementary circuits will define how the different variants impact excitability and thus converge toward an epileptic phenotype. Aim 2 will define hippocampal higher-level circuit perturbations in epileptic mice designed with genome editing to recapitulate the SCN2A-E430A human epileptic encephalopathy. Ex vivo analysis of changes in excitability, synaptic signaling, and network output in the hippocampus will lead to new understanding of how gain-of-function SCN2A variants affect neuronal networks. EEG and depth electrodes will provide spatiotemporal correlate to the in vivo epilepsy phenotype. This proposal will propel the awardee to independence as a physician-scientist by incorporating new expertise in multi-scale modeling of genetic epilepsy, focused relevant didactics, and a diverse career development team specializing in neurodevelopmental and genetic disorders, all in a highly collaborative environment fostering junior faculty development. This award will provide a platform to 1) define variant-specific contributions to epilepsy phenotype in self-limited and intractable epilepsies and 2) investigate how targeted epileptic circuit dysfunction influences circuit output and epilepsy phenotype in future R01-funded independent research.

项目摘要癫痫会影响全球多达1％的人口，在美国影响300万独自的。越来越多的儿科癫痫与离子通道基因的致病变异相关，包括电压门控钠通道基因SCN2A。 Ninds的2020年癫痫研究基准优先确定遗传变异如何引起癫痫和相关的神经发育障碍。 SCN2A变体功能丧失的表现与严重的神经发育障碍和晚期癫痫有关。另一方面，功能良好的SCN2A变体主要具有早发癫痫的表型。编码的钠通道（NAV1.2）早期在兴奋性谷氨酸能神经元中高度表达开发，提供了一个独特的机会，可以检查兴奋性神经元功能障碍如何导致早期发作癫痫。动物和人体组织衍生的神经元模型已为如何Dravet带来了机械洞察力综合征会导致内神经元功能障碍和癫痫。在SCN2A相关疾病中，动物模型阐明了功能丧失如何导致自闭症谱系障碍伴有晚发癫痫。由于缺乏容易可用的疾病模型，对兴奋性神经元功能障碍如何早期有稀疏的机械理解在发育中导致早期发作的癫痫。该建议将利用两个早期发作的癫痫变量具有收敛临床表型但生物物理机制不同的SCN2A。患者衍生的神经元模型和鼠标模型提供了定义多个机械收敛点的机会量表：从单个神经元到影响癫痫表型的神经回路。 AIM 1将决定两个增益 - 功能SCN2A变体，编码错义突变M1879T和E430A，会议提高了兴奋性。不同的机制。 IPSC衍生的神经元的功能分析将孤立和基本回路中的功能分析定义不同变体如何影响兴奋性，从而融合到癫痫表型。 AIM 2意志定义海马高级电路扰动的癫痫小鼠，设计为基因组编辑到概括SCN2A-E430A人类癫痫性脑病。兴奋性变化的体内分析，突触信号传导和海马中的网络输出将导致对功能收益的新了解 SCN2A变体会影响神经元网络。脑电图和深度电极将提供与时空相关的体内癫痫表型。该提议将推动获奖者作为医师科学家的独立性将新的专业知识纳入遗传癫痫的多尺度建模，专注的相关教学和多样的专业知识专门从事神经发育和遗传疾病的职业发展团队，都在高度协作中促进初级教师发展的环境。该奖项将为1）定义特定于变化的平台。在自限和顽固性癫痫中对癫痫表型的贡献，2）研究如何靶向癫痫电路功能障碍会影响未来R01资助的独立的电路输出和癫痫表型研究。