High-throughput interrogation of autism risk genes: from molecules to behavior

项目总结 有1000多个基因与自闭症谱系障碍(ASD)有关，但只有少数几个基因与此有关 在动物模型中被确认为引起ASD相关的表型。了解每个基因是否以及如何 在啮齿动物模型中，单独或联合作用于ASD相关表型是不可行的， 需要大量的时间、费用和人力来生成和描述。我们以前曾利用 线虫作为快速鉴定人类相关基因的最小活体动物模型 神经方面的情况。-线虫显示与ASD相关的表型，包括社会行为 表现为它们在进食时倾向于成堆地聚集在一起。为了迅速洞察109 SFARI基因同源基因在线虫的社会行为中发挥了作用，而不是一次研究一个突变，我们说 研究了一组从世界各地分离出来的遗传上截然不同的野生型菌株。每种毒株都携带一种 这109个ASD风险基因的不同变异组合。我们发现，总体来说，这个数字和 ASD危险基因突变的严重程度与社会行为减少相关。此外，我们发现， 某些自闭症风险基因的突变似乎会导致社交缺陷，因为我们可以促进社交行为 通过用功能版本替换有缺陷的ASD风险基因。我们还发现，同源基因中的突变 导致线虫社会行为增加的基因已经被认为与积极的修改有关 自闭症和威廉姆斯综合征的社会行为。中心假设是线虫将是一种快速和 廉价的模型生物体来确定这大量风险基因的哪些突变组合 导致ASD相关缺陷。总的目标是确定哪些突变和突变组合 ASD风险基因导致ASD相关行为，以及通过什么机制。其基本原理是有一个 迫切需要了解自闭症相关基因突变的体内后果。中环 假说将以三个具体目标进行检验：1)确定ASD风险基因中的哪些自然变异以及如何变异 导致线虫的社会行为和感官整合减少。2)测试哪个和 ASD风险基因的变异如何积极地改变线虫的社会行为和感觉整合。3) 确定ASD患者中不确定意义的遗传变异的机制 影响线虫的社会和感觉整合行为和神经生物学。-研究 在此应用程序中提出的是创新的，因为它使用最低限度的动物模型来执行高吞吐量 自闭症危险基因的体内因果功能分析。这项工作意义重大，因为它将使研究人员 与啮齿动物和人类患者合作，将他们的努力集中在最有希望的自闭症风险基因上。这个 结果将使家庭能够了解未研究或未研究的ASD患者特有的突变是如何 风险基因影响基础神经生物学。归根结底，这些知识有可能指导 未来针对自闭症症状的药物和遗传治疗的发展。

PROJECT SUMMARY Over 1,000 genes have been implicated in autism spectrum disorder (ASD) but only a handful have been confirmed as causing phenotypes related to ASD in animal models. Understanding if and how each gene contributes to ASD-related phenotypes singly or in combination is not feasible with rodent models, which require enormous time, expense, and labor to generate and characterize. We have previously leveraged the nematode C. elegans as a minimum in vivo animal model to quickly characterize genes related to human neurological conditions. ------ C. elegans displays phenotypes with relevance to ASD including social behaviors represented by how they tend to clump together in piles while eating. To quickly gain insight into whether 109 SFARI gene orthologs play a role in social behaviors in C. elegans, rather than study one mutant at a time, we studied a collection of genetically distinct wild-type strains isolated from around the world. Each strain carries a distinct combination of variants in these 109 ASD risk genes. We discovered that overall, the number and severity of mutations in ASD risk genes correlated with decreased social behaviors. Moreover, we found that mutations in certain ASD risk genes appear to cause social deficits, because we could boost social behaviors by replacing defective ASD risk genes with functional versions. We also found that mutations in orthologs of genes that cause increased social behavior in C. elegans have already been implicated in positively modifying social behavior in ASD and Williams syndrome. The central hypothesis is that C. elegans will be a rapid and inexpensive model organism to determine which combinations of mutations in this vast number of risk genes cause ASD-related defects. The overall goal is to determine which mutations and combinations of mutations in ASD risk genes cause ASD-relevant behaviors, and by what mechanisms. The rationale is that there is an urgent need to understand the in vivo consequences of mutations in genes implicated in autism. The central hypothesis will be tested with three specific aims: 1) Identify which and how natural variants in ASD risk genes causally contribute to decreasing social behaviors and sensory integration in C. elegans. 2) Test which and how variants in ASD risk genes positively modify social behaviors and sensory integration in C. elegans. 3) Determine the mechanism by which genetic variants of uncertain significance identified in ASD patients influence social and sensory integration behaviors and neurobiology of C. elegans. ------ The research proposed in this application is innovative because it uses a minimalist animal model to perform high-throughput in vivo causal functional analyses of ASD risk genes. The work is significant because it will allow researchers working with rodents and human patients to focus their efforts on the most promising ASD risk genes. The results will empower families to understand how patient-specific mutations in unstudied or under-studied ASD risk genes influence basic neurobiology. Ultimately, such knowledge has the potential to guide the development of future pharmacological and genetic treatments for the symptoms of ASD.