Developing a zebrafish model of Slc6a1/GAT1 hypofunction and an in vitro assay to identify novel treatments

开发 Slc6a1/GAT1 功能减退的斑马鱼模型和体外测定以确定新的治疗方法

基本信息

批准号：
10041423
负责人：
Christopher McGraw
金额：
$ 20.08万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-15 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10041423
关键词：
Absence Epilepsy Acute Address Affect Antiepileptic Agents Award Biological Assay Biology Boston Brain Brain imaging Cells Chronic Clinical Data Development Disease Drug Screening Epilepsy Equilibrium Fishes Fluorescence Foundations Functional disorder Future GABA Modulators GABA Transporter 1 GABA transporter General Hospitals Generalized Epilepsy Genes Genetic Goals Human Human Cell Line Impairment In Vitro Intractable Epilepsy Investigation Kinetics Knock-out Knowledge Larva Lead Light Massachusetts Measures Mediating Mentorship Modeling Molecular Mutation Myoclonic Astatic Epilepsies Nature Neuraxis Neurons Pathogenesis Patients Pediatric Hospitals Performance Pharmaceutical Preparations Pharmacological Treatment Pharmacology Phenotype Physicians Population Predisposition Property Reporter Reporting Research Research Project Grants Resistance Resources Rodent Role Science Scientist Seizures Signal Transduction Sum Syndrome System Testing Therapeutic Time Work Zebrafish base career career development extracellular gamma-Aminobutyric Acid high throughput screening in vitro Assay in vitro Model in vivo in vivo evaluation infancy inhibitor/antagonist innovation insight interest loss of function loss of function mutation medical schools nervous system development neurophysiology novel novel therapeutics precision medicine programs reuptake screening sensor tiagabine tool uptake

项目摘要

PROJECT SUMMARY/ABSTRACT Epilepsy is a highly prevalent disorder affecting 1% of the world’s population but our understanding of the molecular mechanisms that underlie epilepsy is still incomplete. Deficiency in the function of GABA reuptake mediated by GABA transporter type 1 (GAT1), has been implicated in absence epilepsy and heterozygous mutations in the gene SLC6A1 which encode GAT1 cause myoclonic astatic epilepsy, a generalized seizure disorder with onset in infancy. Several questions remain regarding the pathophysiology of GAT1 hypofunction, including whether the disease can be modeled in zebrafish as it can in rodents and whether the disorder is related to primary constitutive versus secondary developmental consequences of GAT1 hypofunction. Therapeutically, a major question is whether it is possible to identify disease-specific treatments for GABA reuptake deficiency. These questions represent major gaps in knowledge whose answers could inform the timing and nature of treatment for patients with MAE and other generalized epilepsy syndromes involving GAT1 hypofunction. The current proposal will close these gaps by establishing a novel zebrafish model of GAT1 hypofunction to address questions related to the pathogenesis of the disorder and by testing whether a novel in vitro fluorescence-mediated cell-based assay of GAT1 function can be used to identify positive modulators with therapeutic value.The proposed research will provide essential insights into the mechanisms of a generalized epilepsy syndrome related to slc6a1/GAT1 hypofunction and establish platforms for future drug screening and in vivo testing to reverse the pathophysiology of the disorder. The proposal under consideration combines an innovative research project with translational implications, excellent mentorship in science and career development, and extensive institutional resources at Boston Children’s Hospital, Massachusetts General Hospital, and Harvard Medical School, which should facilitate the transition into an independent physician-scientist by the end of the award period. In sum, the proposal provides a framework for a robust independent research program balancing mechanistic and translational investigations of epilepsy. It is well-integrated with the research and clinical interests of the applicant, whose career goal is to become an independent physician scientist with a focus on high-throughput biology and drug screening to understand the mechanisms of epilepsy and to identify corresponding treatments.

项目概要/摘要癫痫是一种非常普遍的疾病，影响着世界上 1% 的人口，但我们对癫痫的认识 GABA 再摄取功能缺陷导致癫痫的分子机制尚不完整。由 1 型 GABA 转运蛋白 (GAT1) 介导，与失神性癫痫和杂合子性癫痫有关编码 GAT1 的基因 SLC6A1 的突变会导致肌阵挛性癫痫，一种全身性癫痫发作婴儿期发病的疾病。关于 GAT1 功能减退的病理生理学仍存在几个问题，包括是否可以像啮齿动物一样在斑马鱼中建立疾病模型，以及该疾病是否与原发性相关 GAT1 功能低下的本质性与继发性发育后果在治疗上是一个主要因素。问题是是否有可能找到针对 GABA 再摄取缺陷的疾病特异性治疗方法。这些问题代表了知识上的重大差距，其答案可以告诉我们这些问题的时间和性质。治疗 MAE 和其他涉及 GAT1 功能减退的全身性癫痫综合征患者。目前的提案将通过建立一种新的 GAT1 功能减退斑马鱼模型来弥补这些差距解决与该疾病发病机制相关的问题，并通过测试是否有新的体外基于 GAT1 功能的荧光介导的细胞测定可用于鉴定正调节剂治疗价值。拟议的研究将为广义的机制提供重要的见解与 slc6a1/GAT1 功能低下相关的癫痫综合征，并为未来的药物筛选和建立平台体内测试以逆转该疾病的病理生理学。正在考虑的提案结合了具有转化意义的创新研究项目，波士顿在科学和职业发展方面提供出色的指导以及广泛的机构资源儿童医院、马萨诸塞州总医院和哈佛医学院，这应该会促进总之，该提案规定，在奖励期结束时转变为独立的医师科学家。平衡机械研究和转化研究的稳健独立研究计划的框架它与申请人的研究和临床兴趣很好地结合在一起，申请人的职业目标是成为一名独立的医师科学家，专注于高通量生物学和药物筛选了解癫痫的机制并确定相应的治疗方法。