Functional Analysis of Rare Variants in Genes Associated with Autism

自闭症相关基因罕见变异的功能分析

基本信息

批准号：
8404053
负责人：
ELLEN J HOFFMAN
金额：
$ 14.66万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-01-03 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8404053
关键词：
Acoustics Adult Architecture Autistic Disorder Axon Behavior Behavior assessment Behavioral Binding Biological Biological Assay Biological Models Biological Process Biology Cell Adhesion Molecules Child Child Development Chromosomes DNA Structure Data Development Developmental Biology Disease Doctor of Philosophy Embryo Emerging Technologies Fertilization Fishes Frameshift Mutation Functional disorder Genes Genetic Genetic Predisposition to Disease Germ-Line Mutation Goals Hour Human Genetics Imagery Immigration Impairment Individual K-Series Research Career Programs Knock-out Knowledge Laboratories Language Larva Lead Maps Medicine Mental disorders Mentors Methods Modeling Molecular Mutation Neural Cell Adhesion Molecules Neuraxis Neurobiology Neurons Neurosciences Orthologous Gene Outcomes Research Pathway interactions Pattern Phenotype Prevalence Process Proteins Psychiatrist Public Health Research Research Personnel Resources Risk Role Social Interaction Susceptibility Gene Synapses System Testing Variant Zebrafish Zinc Fingers autism spectrum disorder base behavior test career contactin disorder risk endonuclease gene discovery gene function genetic manipulation genetic variant genome wide association study human CNTNAP1 protein in vivo innovation insight loss of function migration nervous system development neural circuit neurodevelopment new therapeutic target novel nuclease prepulse inhibition response small molecule social tool zebrafish development

项目摘要

The goal of this career development award is to integrate the genetics of autism spectrum disorders (ASD), developmental neuroscience, and the functional analysis of rare variants, in order to advance our understanding of the basic biological mechanisms underlying ASD. Dr. Hoffman is a board certified, practicing child psychiatrist, who is currently pursuing her PhD in Investigative Medicine in the laboratory of Matthew State, MD, PhD, at the Yale Child Study Center. Her career goal is to become an independent investigator with dual expertise in the genetics of child psychiatric disorders and the neurobiology of vertebrate systems. In this application, she proposes to utilize zebrafish as a novel translational tool that will leverage findings from human genetic studies as a means of advancing our knowledge of the pathophysiology of ASD. Dr. Hoffman will gain this expertise through the combined guidance of her primary mentor, Dr. State, a leader in ASD genetics, and co-mentor, Antonio Giraldez, PhD, an expert in zebrafish development, which will promote the establishment of this innovative approach to investigating the role of ASD susceptibility genes in neural circuit formation. The objective of this research is to elaborate basic mechanisms of ASD by investigating the function of the ASD risk gene, Contactin Associated Protein-2 (CNTNAP2) in neural development, and to determine how sequence variants in this gene identified in individuals with ASD disrupt its function. Using the emerging technology of zinc finger nucleases, which have superior accuracy over morpholinos, Dr. Hoffman induced targeted germline mutations in the two zebrafish CNTNAP2 genes, CNTNAP2a and 2b. To our knowledge, this is the first zebrafish knockout of an ASD risk gene generated by this method. Our hypothesis is that CNTNAP2a/2b double knockouts will display reproducible morphological and/or behavioral phenotypes that will yield important insights into the function of CNTNAP2 in neural development. This hypothesis will be tested by pursuing these aims: 1) Generate double knockouts of the two zebrafish CNTNAP2 genes by crossing fish carrying germline mutations; 2) Identify quantifiable morphological and/or behavioral phenotypes in CNTNAP2 double knockouts; and 3) Characterize the ability of the human CNTNAP2 gene with rare sequence variants found in individuals with ASD to reverse the phenotypes. Our rationale for this approach is that the development of an in vivo system to rapidly assess the functional consequences of rare genetic variants is the crucial next step in understanding the biology of ASD. Dr. Hoffman has assembled an outstanding team of mentors and collaborators, including pioneers in zebrafish neural circuit analysis and behavioral phenotyping, as this system is anticipated to provide unique insights into the role of CNTNAP2 in the neural circuitry underlying simple behaviors. The extensive resources of the Yale Child Study Center, together with her formal plan for didactics in neuroscience and developmental biology, will further support Dr. Hoffman¿s goal of elucidating the molecular and cellular mechanisms of ASD.

该职业发展奖的目标是整合自闭症谱系障碍（ASD），发育神经科学和稀有变体功能分析的遗传学，以促进我们对ASD基本生物学机制的理解。霍夫曼博士是董事会认证，实践儿童精神科医生，目前正在耶鲁大学儿童学习中心在马修州马修州立大学马修州实验室攻读研究医学博士学位。她的职业目标是成为儿童精神疾病遗传学和脊椎动物系统神经生物学遗传学双重专家的独立研究者。在这个她建议将斑马鱼用作一种新型的翻译工具，该工具将利用人类遗传研究的发现，作为提高我们对ASD病理生理学知识的一种手段。 Hoffman博士将通过其主要导师，ASD遗传学领导者State博士和Zebrafish Development的专家Antonio Giraldez博士的联合指导来获得这一专家，该博士将促进这种创新的方法来研究ASD易感遗传在神经循环形成中的作用。这项研究的目的是通过研究ASD风险基因的功能，接触蛋白相关的蛋白-2（CNTNAP2）在神经元发育中的功能来详细阐述ASD的基本机制，并确定ASD患者中鉴定在该基因中的序列变体如何破坏其功能。霍夫曼博士利用具有优于吗啡的锌指核素的新兴技术，在两个斑马鱼CNTNAP2基因CNTNAP2A和2B中诱导了靶向的种系突变。据我们所知，这是该方法产生的ASD风险基因的第一个斑马鱼敲除。我们的假设是CNTNAP2A/2B双敲除将显示可再现的形态学和/或行为表型，这些表型将对CNTNAP2在神经发育中的功能产生重要的见解。该假设将通过追求这些目标来检验：1）通过越过携带种系突变的鱼类来产生两个斑马鱼CNTNAP2基因的双重敲除； 2）确定CNTNAP2双敲除中的可量化形态和/或行为表型； 3）表征人类CNTNAP2基因具有在ASD个体中发现的稀有序列变异的能力，可以逆转表型。我们对这种方法的理由是，开发体内系统以快速评估稀有遗传变异的功能后果是了解ASD生物学的关键下一步。霍夫曼博士召集了一支杰出的导师和合作者团队，包括斑马鱼神经电路分析和行为表型的先驱，因为该系统预计该系统将为CNTNAP2在基本的简单行为基础的神经电路中的作用提供独特的见解。耶鲁儿童研究中心的广泛资源以及她在神经科学和发育生物学领域的教学计划的正式计划，将进一步支持霍夫曼博士阐明ASD的分子和细胞机制的目标。