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.

项目概要 自闭症谱系障碍（ASD）是由环境和遗传因素共同引起的，遗传因素 贡献估计为 60-80%。已经发现了数十种增加自闭症谱系障碍风险的基因，其中大多数是基于 但这些突变预计仅占 ASD 病例的 15-20%。因此， 据预测，自闭症谱系障碍的大部分遗传因素是由常见和罕见的遗传变异造成的，但是 很少有这样的基因被发现。最近，利用全基因组测序，我们报告了全基因组 超过 60 个 ASD 风险基因的证据，其中 26 个是 ASD 的新基因，信号来自遗传和新生 蛋白质截短或错义突变。大多数这些基因的功能是未知的，因此一个至关重要的和 下一步必要的是使用模型探索它们对神经发育和神经元功能的影响 生物。目前将遗传风险因素转化为表型、机制和疗法的步伐是 部分受到体内哺乳动物模型系统效率低下的限制，这使得它们对于创建 并对大量突变株系进行行为测试。在这里，我们利用斑马鱼，它占据了一个利基市场 作为一种脊椎动物模型，具有适合体内筛选和机制理解的特征，包括 脊椎动物大脑保守但较小，与 ASD 相关的行为以及相对于哺乳动物的成本效益 模型。虽然斑马鱼无法重现自闭症谱系障碍 (ASD)，并且在模拟人类疾病方面也存在局限性，但 新兴文献支持这样一种观点，即它是研究基因功能的有用模型，这些基因有助于 自闭症谱系障碍风险。我们将加快机械化进程，而不是一次评估一个自闭症谱系障碍（ASD）风险基因。 通过高通量测定和分析来理解。在家长资助中，我们建议使用全脑 钙成像研究斑马鱼 ASD 风险基因突变体幼虫期的神经元网络特性 发展。这个多样性补充应用程序描述了一个实验性和概念性的职业 研究生发展计划，其实验目标是（1）建立全脑系统 在呈现虚拟社交线索期间对幼年斑马鱼进行钙成像，以及（2）使用该系统来识别 斑马鱼 ASD 风险基因突变体与野生型对照相比的神经元网络特性 社交线索。该实验计划通过表征响应中的大脑状态来直接与家长资助相关 斑马鱼在幼年发育阶段首次表现出社交行为。这些 实验与父资助中描述的实验是分开的，但又相互协同。在一起， 家长资助和多样性补充有可能确定解释神经元机制 在含有 ASD 风险基因突变的斑马鱼中观察到的行为表型。