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PCDH19-related Epilepsy-Human Genotype-Phenotype Studies Lead to Zebrafish Studies

PCDH19相关癫痫-人类基因型-表型研究引发斑马鱼研究

基本信息

批准号：
10308052
负责人：
Annapurna Poduri
金额：
$ 38.72万
依托单位：
BOSTON CHILDREN'S HOSPITAL
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：
2017-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10308052
关键词：
Address Adhesions Affect Alleles Animal Model Antiepileptic Agents Behavioral Biological Models Biology Brain CRISPR/Cas technology Cadherin Domain Cell Adhesion Cell-Cell Adhesion Cells Childhood Cognitive Convulsants Cortical Dysplasia Cytoplasmic Tail Data Defect Development Disease Drug Screening Electrophysiology (science)Epilepsy Fathers Female Functional disorder Genes Genetic Genotype Germ-Line Mutation Human Human Genetics Hyperthermia Individual Inheritance Patterns Inherited Intellectual functioning disability Label Larva Lead Link Location Mediating Microelectrodes Missense Mutation Modeling Mosaicism Mothers Mutation Nervous system structure Neuraxis Neurons PCDH19 female-limited epilepsy Parents Patients Phenotype Play Point Mutation Predisposition Proteins Publishing Refractory Registries Reporting Research Resources Role Seizures Series Severities Study models System Testing Transgenic Organisms Translational Research X Inactivation Zebrafish autism spectrum disorder base cell motility cell type drug discovery excitatory neuron extracellular gene discovery genome editing genotyped patients high throughput analysis human model in vivo inhibitory neuron insight lifetime risk loss of function male mutant nervous system disorder neurodevelopment novel novel strategies patch clamp precision medicine profiles in patients protocadherin 19 response

项目摘要

Epilepsy is a common condition, affecting 1 in 26 individuals, for which the role of genetics is well recognized. Since its discovery in 2008, PCDH19 has been amongst the most prominent single gene causes of epilepsy. Mutations in PCDH19 cause X-linked “female limited epilepsy,” with refractory childhood-onset seizures, intellectual disability, and autism. PCDH19 encodes a central nervous system protocadherin predicted to mediate cell adhesion, but its role in neurodevelopment is not established. PCDH19 mutations can be de novo or inherited from mildly affected/unaffected mothers or, curiously, unaffected fathers. The female predominance is hypothesized to be due to ‘obligate mosaicism.’ Reports of rare symptomatic mosaic males are consistent with this hypothesis. However, recently noted subtle behavioral features in ‘carrier’ fathers suggest that a mosaic state is not required for all phenotypic manifestations. We will address, using these unusual genetics as a clue, the functional role of PCDH19 and its role in epilepsy. We will conduct genotype- phenotype studies to understand the importance of mutation location, type, and inheritance on PCDH19- related dysfunction. We will then harness a carefully curated set of human PCDH19 mutations, from the PCDH19 Registry that we founded, into zebrafish mechanistic models. There are currently no established animal models of PCDH19-induced epilepsy. Zebrafish represent an established vertebrate model system with genetic tractability and identifiable seizures. Our preliminary data from CRISPR/Cas9 genome-edited zebrafish suggest that loss of function of pcdh19 results in seizures. With these models, we will study the cell types involved and the specific mechanisms by which loss or alteration of pcdh19 results in disease. We hypothesize that the location and type of PCDH19 mutation correlates with phenotypic severity in humans and that zebrafish models based on human mutations will display seizures. We thus pursue a novel approach to study the mechanisms involved in PCDH19 dysfunction, with models based on mutations that we have accrued through our Registry. We will aim to correlate the severity of PCDH19-related phenotypes with location and type of patient mutations; establish pcdh19 zebrafish models based on human mutations and characterize their epilepsy phenotypes; and identify the effects of mutations on cell migration, adhesion, and excitability in mutant pcdh19 zebrafish. Our research will develop the first in vivo animal models for PCDH19-related epilepsy. Through our zebrafish models, we will investigate the neurodevelopmental role of PCDH19 and develop drug screens for PCDH19- related epilepsy in the emerging era of precision medicine. The broader impacts of this study are that it will (1) provide insight into protocadherin-related epilepsies, (2) establish a human-to-zebrafish paradigm for translational research in genetic epilepsy, and (3) inform the modeling of other mosaic neurological disorders.

癫痫是一种常见的疾病，每26个人中就有1个受影响，遗传学的作用得到了充分的认可。自2008年发现以来，PCDH 19一直是癫痫最重要的单基因病因之一。 PCDH 19的突变导致X连锁的“女性局限性癫痫”，伴随难治性儿童期发作癫痫发作，智力残疾和自闭症。PCDH 19编码一种中枢神经系统原钙粘蛋白，介导细胞粘附，但其在神经发育中的作用尚未确定。PCDH 19突变可以是新生的，或者遗传自轻度受影响/未受影响的母亲，或者，奇怪的是，未受影响的父亲。女假设优势是由于“专性镶嵌”。报告罕见的症状马赛克男性与这个假设是一致的。然而，最近注意到“携带者”父亲的微妙行为特征，表明镶嵌状态不是所有表型表现所必需的。我们将利用这些不寻常的遗传学作为线索，PCDH 19的功能作用及其在癫痫中的作用。我们会进行基因分型- 表型研究，以了解突变位置，类型和遗传对PCDH 19- 相关功能障碍。然后，我们将利用一组精心策划的人类PCDH 19突变， PCDH 19注册表，我们建立，到斑马鱼的机械模型。目前还没有建立PCDH 19诱导癫痫的动物模型。斑马鱼代表了建立了具有遗传易处理性和可识别癫痫发作的脊椎动物模型系统。我们的初步数据来自CRISPR/Cas9基因组编辑的斑马鱼的研究表明，pcdh 19功能的丧失导致癫痫发作。与在这些模型中，我们将研究所涉及的细胞类型和特定的机制，通过这些机制， PCDH 19导致疾病。我们假设PCDH 19突变的位置和类型与基于人类突变的斑马鱼模型将显示癫痫发作。因此，我们寻求一种新的方法来研究PCDH 19功能障碍的机制，基于我们通过注册表积累的突变的模型。我们的目标是将 PCDH 19相关表型的严重程度与患者突变的位置和类型;建立pcdh 19斑马鱼基于人类突变的模型，并描述其癫痫表型;并确定突变体pcdh 19对斑马鱼细胞迁移、粘附和兴奋性的影响。我们的研究将开发出第一个PCDH 19相关癫痫的体内动物模型。通过我们的斑马鱼模型，我们将研究PCDH 19的神经发育作用，并开发PCDH 19的药物筛选，在精准医疗的新兴时代与癫痫相关。这项研究的更广泛的影响是，它将（1）提供对原钙粘蛋白相关癫痫的深入了解，（2）建立一个人类到斑马鱼的范例，遗传性癫痫的转化研究，和（3）告知其他马赛克神经系统疾病的建模。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

DOI：
10.1111/epi.14003
发表时间：
2018-03
期刊：
Epilepsia
影响因子：
5.6
作者：
Smith L;Singhal N;El Achkar CM;Truglio G;Rosen Sheidley B;Sullivan J;Poduri A
通讯作者：
Poduri A

Cortical Excitability, Synaptic Plasticity, and Cognition in Benign Epilepsy With Centrotemporal Spikes: A Pilot TMS-EMG-EEG Study.