High-throughput modeling of autism risk genes using zebrafish

使用斑马鱼进行自闭症风险基因的高通量建模

• 批准号: 10478187
• 负责人: DANIEL H GESCHWIND
• 金额: $ 75.66万
• 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/10478187
• 关键词: Address; Animal Model; Autopsy; Behavior; Behavioral; Behavioral Model; Biological Assay; Biological Models; Brain; Cells; Communities; Data; Development; Disease; Disease model; Drug Screening; Gene Expression; Genes; Genetic; Goals; Human; Image; Individual; Inherited; Lead; Literature; Methods; Missense Mutation; Modeling; Molecular; Mutation; Neurons; Orthologous Gene; Pathway Analysis; Pathway interactions; Pharmacology; Phenotype; Proteins; Publishing; RNA Splicing; Reagent; Reporting; Resources; Risk; Role; Seizures; Signal Transduction; Sleep; Sleep disturbances; Social Behavior; Spliced Genes; Startle Reaction; System; Testing; Time; Tissue-Specific Gene Expression; Translating; Variant; Vertebrates; Zebrafish; autism spectrum disorder; base; behavior test; behavioral phenotyping; brain behavior; cohort; comorbidity; cost; cost effectiveness; de novo mutation; disorder risk; experimental study; gene function; gene network; genetic risk factor; genetic testing; genome sequencing; genome wide association study; genome-wide; habituation; high throughput analysis; high throughput screening; high-throughput drug screening; imaging approach; improved; in vivo; molecular phenotype; mutant; neurodevelopment; novel; novel therapeutic intervention; novel therapeutics; null mutation; relative effectiveness; risk variant; screening; social; transcriptomics; translational goal; whole genome

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 unique niche as a vertebrate model with features amenable to both in vivo screening and mechanistic understanding, including ex utero development, transparency, small size, rapid development, a conserved yet relatively simple 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 Specific Aim 1 we will generate null mutations in the zebrafish orthologs of 24 high confidence, novel, genome-wide significant ASD risk genes, and systematically test each mutant for neurodevelopmental, behavioral, neuronal network, and transcriptomic phenotypes. In Specific Aim 2, we will use transcriptomic analyses, at the whole brain and single cell levels, to integrate ASD risk genes into functional networks, and test for convergence across genes and species, including ASD post mortem brain. We will also test for functional associations among behavioral phenotypes that are often co-morbid in ASD, such as disrupted sleep and social behavioral deficits. In Specific Aim 3 we will perform mechanistic studies to understand how mutation of specific ASD-risk genes leads to phenotypes. This project will efficiently and cost-effectively create and characterize vertebrate animal models for a large number of novel ASD risk genes. These animal models will be a valuable resource for the community, particularly for large-scale in vivo drug screens to identify new therapies for ASD.

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，它占据了独特 利基作为一种脊椎动物模型，具有适合体内筛查和机械理解的特征， 包括前子宫开发，透明度，小规模，快速发展，保守但相对简单的 脊椎动物的大脑，与ASD相关的行为以及与哺乳动物模型相关的成本效益。而 斑马鱼不能概括ASD，并且对建模人类疾病有局限性，这是一种新兴文献 支持以下观点：研究有助于ASD风险的基因功能是一个有用的模型。相当 比一次评估ASD风险基因一个人，我们将通过 高通量测定和分析。在特定目标1中，我们将在斑马鱼直系同源物中产生无效突变 24个高置信度，新型，全基因组明显的ASD风险基因，并系统地测试每个突变体的 神经发育，行为，神经元网络和转录组表型。在特定的目标2中，我们将 在整个大脑和单细胞水平上使用转录组分析将ASD风险基因整合到功能上 网络，以及跨基因和物种（包括ASD Mortem大脑）的收敛。我们也会 测试通常在ASD中合并的行为表型之间的功能关联，例如破坏 睡眠和社会行为缺陷。在特定目标3中，我们将进行机械研究，以了解如何 特定ASD风险基因的突变导致表型。该项目将有效地创建 并表征了大量新型ASD风险基因的脊椎动物模型。这些动物模型 对于社区而言，将是一个宝贵的资源，特别是对于大规模的体内药物屏幕来识别新的 ASD的疗法。