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

基本信息

批准号：
10264069
负责人：
DANIEL H GESCHWIND
金额：
$ 75.68万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
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/10264069
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 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 whole genome

项目摘要

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系进行行为测试。在这里，我们利用斑马鱼，它占据了独特的地位利基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，包括子宫外发育、透明、体积小、发育快、保守但相对简单脊椎动物大脑、与 ASD 相关的行为以及相对于哺乳动物模型的成本效益。虽然斑马鱼无法重现自闭症谱系障碍，并且在模拟人类疾病方面存在局限性，这是一项新兴文献支持这样的观点，即它是研究导致 ASD 风险的基因功能的有用模型。相当与一次评估一个自闭症谱系障碍（ASD）风险基因相比，我们将通过以下方式加速机制理解的进展：高通量测定和分析。在特定目标 1 中，我们将在斑马鱼直向同源物中生成无效突变 24 个高置信度、新颖、全基因组显着的 ASD 风险基因，并系统地测试每个突变体神经发育、行为、神经元网络和转录组表型。在具体目标 2 中，我们将在全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中网络，并测试跨基因和物种的收敛性，包括 ASD 死后大脑。我们也会测试自闭症谱系障碍 (ASD) 中常见的行为表型之间的功能关联，例如紊乱睡眠和社交行为缺陷。在具体目标 3 中，我们将进行机制研究以了解如何特定 ASD 风险基因的突变会导致表型。该项目将有效且具有成本效益地创造并表征脊椎动物模型中大量新的 ASD 风险基因。这些动物模型将成为社区的宝贵资源，特别是对于大规模体内药物筛选以识别新的药物自闭症谱系障碍 (ASD) 的治疗方法。