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

• 批准号: 8595337
• 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
• 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/8595337
• 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; rare variant; response; risk variant; small molecule; social; tool; zebrafish development

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.

这个职业发展奖的目标是整合自闭症谱系障碍(ASD)的遗传学、发育神经科学和罕见变异的功能分析，以促进我们对ASD基本生物学机制的理解。霍夫曼博士是一名委员会认证的执业儿童精神病学家，目前正在耶鲁儿童研究中心马修州立大学的实验室攻读调查医学博士学位。她的职业目标是成为一名拥有儿童精神障碍遗传学和脊椎动物系统神经生物学双重专业知识的独立研究员。在这 关于斑马鱼的应用，她建议利用斑马鱼作为一种新的翻译工具，利用人类基因研究的结果作为促进我们对自闭症病理生理学的知识的一种手段。霍夫曼博士将通过她的主要导师、ASD遗传学领袖State博士和共同导师Antonio Giraldez博士的共同指导获得这些专业知识，Antonio Giraldez博士是斑马鱼发育方面的专家，这将促进建立这一创新方法，以研究ASD易感基因在神经回路形成中的作用。 本研究的目的是通过研究ASD风险基因--联系蛋白相关蛋白-2(CNTNAP2)在神经发育中的功能，阐明ASD的基本机制，并确定在ASD患者中发现的该基因的序列变异如何破坏其功能。霍夫曼博士利用锌指核酸酶这一新兴技术，在斑马鱼的CNTNAP2基因CNTNAP2a和2b中诱导了有针对性的种系突变。锌指核酸酶的准确性高于吗啉。据我们所知，这是通过这种方法产生的ASD风险基因的第一个斑马鱼基因敲除。我们的假设是，CNTNAP2a/2b双敲除将显示出可重复的形态和/或行为表型，这将对CNTNAP2在神经发育中的功能产生重要的见解。这一假说将通过追求以下目标来检验：1)通过杂交携带种系突变的鱼类来产生两个斑马鱼CNTNAP2基因的双敲除；2)确定CNTNAP2双敲除中可量化的形态和/或行为表型；以及3)表征在ASD患者中发现的具有罕见序列变体的人CNTNAP2基因逆转表型的能力。我们这种方法的基本原理是，开发一个体内系统来快速评估罕见基因变异的功能后果是理解ASD生物学的关键下一步。 霍夫曼博士组建了一支由导师和合作者组成的杰出团队，其中包括斑马鱼神经回路分析和行为表型的先驱，因为该系统有望为CNTNAP2在简单行为背后的神经回路中所扮演的角色提供独特的见解。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正式教学计划，将进一步支持霍夫曼·S博士阐明自闭症分子和细胞机制的目标。