Functional evaluation of catastrophic childhood epilepsy genes in zebrafish

斑马鱼灾难性儿童癫痫基因的功能评估

基本信息

批准号：
9905567
负责人：
Scott C Baraban
金额：
$ 34.67万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9905567
关键词：
3-Dimensional Animal Model Antiepileptic Agents Basic Science Behavior Behavior monitoring Behavioral Benchmarking Biological Assay Brain Brain imaging CRISPR/Cas technology Calcium Cardiovascular Diseases Cell Culture Techniques Child Clinical Clustered Regularly Interspaced Short Palindromic Repeats Communities Complex Consumption DNA Sequence Alteration Databases Diabetes Mellitus Diagnosis Disease Early Infantile Epileptic Encephalopathy Electrophysiology (science)Epilepsy Etiology Evaluation Exhibits Functional disorder Funding Gene Mutation Generations Genes Genetic Genetic Diseases Goals Home environment Human Human Genetics Image International Laboratories Lead Life Locomotion Malignant Neoplasms Methodology Methods Mission Modeling Modification Mutant Strains Mice National Institute of Neurological Disorders and Stroke Neurons Oregon Patients Phenotype Pre-Clinical Model Research Resistance Resistance profile Resources Seizures Speed System Techniques Time United States National Institutes of Health Universities Vertebrates Vision Zebrafish autism spectrum disorder childhood epilepsy design disability drug discovery effective therapy gene function genome wide association study improved in vivo induced pluripotent stem cell insight loss of function loss of function mutation multidisciplinary mutant neural network next generation sequencing open source patient population pre-clinical precision medicine preclinical development relating to nervous system side effect stem cell technology tool

项目摘要

Project Summary/Abstract Children with catastrophic epilepsies develop intractable forms of epilepsy with severe intellectual and behavioral disabilities, including autism. Many of these children are diagnosed, early in life, with genetic forms of epilepsy. With recent advances in gene sequencing approximately 60 loss-of-function (LOF) single-gene mutations have been identified in this patient population. Zebrafish LOF mutants designed to represent these human genetic conditions would provide valuable tools for elucidating basic disease mechanisms and drug discovery. Methods such as the CRISPR/Cas9 system (clustered regularly interspaced short palindromic repeat/Cas9) now allow for rapid and efficient modification of endogenous genes in a range of animal models, including zebrafish (Danio rerio). Using this technique, we recently developed and characterized a stxbp1 mutant zebrafish line representing children with early infantile epileptic encephalopathy with burst suppression. In this proposal, we will generate pre-clinical mutant zebrafish lines for all known human LOF mutations using CRISPR/Cas9 gene editing techniques (Aim I). To determine whether these mutants exhibit functional phenotypes (including epilepsy), we will carefully monitor behavior and neural activity using unique assays and methodologies developed in our laboratory (Aim II). Our strategy includes (i) detailed analysis of behavior using automated locomotion tracking and high-speed imaging, (ii) recording brain activity using in vivo electrophysiology techniques and (iii) brain-wide calcium imaging of neural networks. Zebrafish lines with epileptic phenotypes will also be screened against a small panel of existing antiepileptic drugs. All zebrafish mutant lines generated and carefully characterized here will be made available to the scientific community via the NINDS-supported Zebrafish International Resource Center in Oregon. Both as a resource and offering insight into functional consequences of these single gene mutations, our proposal establishes a comprehensive zebrafish-based precision medicine approach targeted to monogenic epilepsy disorders seen primarily in children.

项目概要/摘要患有灾难性癫痫的儿童会发展为顽固性癫痫，伴有严重的智力和智力障碍。行为障碍，包括自闭症。这些儿童中的许多人在生命早期就被诊断出患有遗传形式癫痫症。随着基因测序的最新进展，大约有 60 个功能丧失 (LOF) 单基因在该患者群体中已发现突变。斑马鱼 LOF 突变体旨在代表这些人类遗传条件将为阐明基本疾病机制和药物提供有价值的工具发现。诸如 CRISPR/Cas9 系统（聚类规则间隔短回文）等方法重复/Cas9）现在允许在一系列动物模型中快速有效地修饰内源基因，包括斑马鱼（Danio rerio）。使用这种技术，我们最近开发并表征了 stxbp1 代表具有爆发抑制的早期婴儿癫痫性脑病儿童的突变斑马鱼系。在本提案中，我们将使用以下方法针对所有已知的人类 LOF 突变生成临床前突变斑马鱼系 CRISPR/Cas9 基因编辑技术（目标 I）。为了确定这些突变体是否表现出功能表型（包括癫痫），我们将使用独特的分析方法仔细监测行为和神经活动我们实验室开发的方法（目标 II）。我们的策略包括 (i) 详细的行为分析使用自动运动跟踪和高速成像，（ii）使用体内记录大脑活动电生理学技术和（iii）神经网络的全脑钙成像。斑马鱼线与还将针对一小部分现有抗癫痫药物筛选癫痫表型。所有斑马鱼此处生成并仔细表征的突变系将通过以下方式提供给科学界 NINDS 支持的位于俄勒冈州的斑马鱼国际资源中心。既作为资源又作为产品深入了解这些单基因突变的功能后果，我们的建议建立了针对单基因癫痫病的基于斑马鱼的综合精准医学方法主要发生于儿童。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Phenotypic analysis of catastrophic childhood epilepsy genes.

DOI：
10.1038/s42003-021-02221-y
发表时间：
2021-06-03
期刊：
Communications biology
影响因子：
5.9
作者：
Griffin A;Carpenter C;Liu J;Paterno R;Grone B;Hamling K;Moog M;Dinday MT;Figueroa F;Anvar M;Ononuju C;Qu T;Baraban SC
通讯作者：
Baraban SC