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 估计占60- 80%。已经确定了数十种增加ASD风险的基因，其中大部分基于 新生突变，但预计这些突变仅占ASD病例的15-20%。因此 预测ASD的大部分遗传贡献来自常见和罕见的遗传变异，但是 很少有这样的基因被鉴定出来。最近，利用全基因组测序， 有证据表明有超过60个ASD风险基因，其中26个是ASD的新基因，其信号来自遗传和新生 蛋白质截短或错义突变。这些基因中的大多数的功能是未知的，所以一个关键的， 必要的下一步是使用模型探索它们对神经发育和神经元功能的影响 有机体目前将遗传风险因素转化为表型、机制和治疗方法的速度是 部分受限于体内哺乳动物模型系统的效率低下，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， 作为一种脊椎动物模型，其特征适合于体内筛选和机理理解，包括 一个保守的小脊椎动物的大脑，行为相关的ASD，和成本效益相对于哺乳动物 模型虽然斑马鱼不能重现ASD，并且在模拟人类疾病方面存在局限性，但研究人员认为， 新兴的文献支持这一概念，即它是一个有用的模型，研究基因的功能，有助于 ASD风险。与其一次评估一个ASD风险基因，我们将加速向机械化的方向发展。 通过高通量测定和分析来理解。在父母补助金中，我们建议使用全脑 钙成像研究斑马鱼ASD风险基因突变体在幼鱼阶段的神经网络特性 发展这种多样性补充申请描述了一个实验性和概念性的职业生涯 一个研究生的发展计划，其实验目标是（1）建立一个系统， 钙成像的幼年斑马鱼在呈现虚拟社会线索，和（2）使用这个系统，以确定 与野生型对照相比，斑马鱼ASD风险基因突变体的神经元网络特性响应于 社会线索这个实验计划直接涉及到父母补助金的特点大脑状态的反应 社会线索在青少年阶段的发展，当斑马鱼第一次显示社会行为。这些 实验与母基金中描述的实验是分开的，但又是协同的。统称 父母补助金和多样性补助金有可能确定神经元机制，解释 在斑马鱼中观察到的行为表型包含ASD风险基因的突变。