High-throughput Modeling of Autism Risk Genes using Zebrafish - DIVERSITY SUPPLEMENT

使用斑马鱼对自闭症风险基因进行高通量建模 - 多样性补充

• 批准号: 10818861
• 负责人: DANIEL H GESCHWIND
• 金额: $ 9.4万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10818861
• 关键词: Acceleration; Address; Adolescent; Age; Bachelor' s Degree; Behavior; Behavioral; Biological Models; Biological Process; Brain; Calcium; Child; Cognitive Science; Communities; Computer Analysis; Cues; Data; Data Set; Development; Development Plans; Disease; Disease model; Doctor of Philosophy; Education; Eligibility Determination; Ensure; Feedback; Fellowship; Fishes; Genes; Genetic; Goals; Grant; Health; Human; Image; Individual; Inherited; Journals; Knowledge; Latino; Literature; Mentors; Methods; Missense Mutation; Modeling; Mus; Mutation; Neurons; Neurosciences; Oral; Parents; Phenotype; Property; Protein Truncation; Publishing; Reporting; Research; Risk; Role; Scientific Advances and Accomplishments; Scientist; Signal Transduction; Social Behavior; System; Techniques; Testing; Time; Training; Translating; United States National Institutes of Health; Variant; Writing; Zebrafish; analytical method; autism spectrum disorder; behavior test; behavioral phenotyping; career; career development; complex data; computer science; cost effectiveness; de novo mutation; design; disorder risk; experience; experimental study; gene function; genetic risk factor; genome sequencing; genome-wide; graduate student; high throughput screening; imaging study; improved; in vivo; laboratory experiment; member; model organism; mutant; nervous system disorder; neurodevelopment; novel; nutrition; parent grant; pre-doctoral; programs; relative effectiveness; response; risk variant; screening; skills; social; success; symposium; synergism; tool; virtual; whole genome

PROJECT SUMMARY Autism spectrum disorder (ASD) is caused by both environmental and genetic factors, with the genetic contribution estimated at 60-80%. Dozens of genes that increase risk for ASD have been identified, most based on de novo mutations, but these mutations are predicted to account for only 15-20% of ASD cases. Thus, the majority of the genetic contribution to ASD is predicted to result from common and rare inherited variation, but few such genes have been identified. Recently, using whole genome sequencing, we reported genome wide evidence for >60 ASD risk genes, 26 of them novel for ASD, with signals derived from inherited and de novo protein truncating or missense mutations. The functions of most of these genes are unknown, so a crucial and necessary next step is to explore their impact on neurodevelopment and neuronal function using a model organism. The current pace of translating genetic risk factors into phenotypes, mechanisms and therapies is limited in part by inefficiencies with in vivo mammalian model systems, which makes them impractical for creating and behaviorally testing large numbers of mutant lines. Here, we leverage the zebrafish, which occupies a niche as a vertebrate model with features amenable to both in vivo screening and mechanistic understanding, including a conserved yet small vertebrate brain, behaviors relevant to ASD, and cost-effectiveness relative to mammalian models. While the zebrafish cannot recapitulate ASD and has limitations for modeling a human disorder, an emerging literature supports the notion that it is a useful model to study the functions of genes that contribute to ASD risk. Rather than assess ASD-risk genes one at a time, we will accelerate progress towards mechanistic understanding via high-throughput assays and analyses. In the parent grant, we proposed to use whole-brain calcium imaging to study neuronal network properties of zebrafish ASD risk gene mutants at the larval stage of development. This diversity supplement application describes an experimental and conceptual career development plan for a graduate student whose experimental goals are to (1) establish a system for brain-wide calcium imaging of juvenile zebrafish during presentation of virtual social cues, and (2) use this system to identify neuronal network properties of zebrafish ASD risk gene mutants compared to wild-type controls in response to social cues. This experimental plan directly relates to the parent grant by characterizing brain states in response to social cues at the juvenile stage of development, when zebrafish first show social behaviors. These experiments are separate from, yet synergize with, the experiments described in the parent grant. Together, the parent grant and diversity supplement have the potential to identify neuronal mechanisms that explain the behavioral phenotypes observed in zebrafish that contain mutations in ASD risk genes.

项目摘要 自闭症谱系障碍（ASD）是由环境和遗传因素引起的，遗传因素 贡献估计为60-80％。已经确定了增加增加ASD风险的数十个基因，大多数基于 关于从头突变，但预计这些突变仅占ASD病例的15-20％。因此， 预计大多数对ASD的遗传贡献是由常见和罕见的遗传变异引起的，但 很少有这样的基因被鉴定出来。最近，使用整个基因组测序，我们报道了基因组范围 > 60个ASD风险基因的证据，其中26个是ASD的新颖的，其信号来自遗传和从头开始 蛋白质截断或错义突变。大多数这些基因的功能都是未知的，因此是至关重要的 下一步的必要下一步是使用模型探索其对神经发育和神经元功能的影响 生物。当前将遗传危险因素转化为表型，机制和疗法的速度是 一部分受体内哺乳动物模型系统效率低下的限制，这使得它们在创建中不切实际 并在行为上测试大量突变线。在这里，我们利用斑马鱼，斑马鱼 作为具有体内筛查和机械理解的特征的脊椎动物模型，包括 保守而又小的脊椎动物大脑，与ASD相关的行为以及相对于哺乳动物的成本效益 型号。尽管斑马鱼不能概括ASD，并且对人类疾病建模有局限性，但 新兴文献支持以下观念：研究有助于有助于的基因功能的有用模型 ASD风险。而不是一次评估ASD风险基因，我们将加速进度 通过高通量测定和分析理解。在父母赠款中，我们建议使用全脑 钙成像研究在幼虫阶段的斑马鱼ASD风险基因突变体的神经元网络特性 发展。这种多样性补充应用描述了实验和概念的职业 一个研究生的开发计划，其实验目标是（1）建立一个大脑范围的系统 少年斑马鱼的钙成像在虚拟社交提示期间，以及（2）使用此系统识别 斑马鱼ASD风险基因突变体的神经元网络特性与野生型对照相比 社会提示。该实验计划直接与父母的赠款有关，以表征大脑状态以响应 在少年发展阶段，斑马鱼首次表现出社会行为时。这些 实验与父授予授予中描述的实验分开但协同作用。在一起， 父母的赠款和多样性补充有可能识别解释神经元机制 在斑马鱼中观察到的行为表型，其中包含ASD风险基因中的突变。