Characterizing the Genetics of FASD in Complementary Mouse and Fish Models

项目概要/摘要 妊娠期酒精暴露是导致出生缺陷的主要环境原因， 神经系统功能障碍虽然酒精对大脑和面部的影响已经被探索得相当多， 广泛地说，发育期接触乙醇的后果有相当大的差异。一些 这种差异是由于暴露时间和暴露量的差异或营养因素造成的。然而，即使当 尽管控制了这些因素，但很明显，并非所有在发育过程中接触乙醇的人都会受到影响。 同样，越来越清楚的是，遗传因素在胎儿酒精中起着非常重要的作用， 谱系障碍（FASD）。从历史上看，阐明这些介导风险或弹性的遗传因素， 相对缓慢，其特征是人类关联研究或动物数量性状基因座作图 模型最近，我们应用了下一代测序技术和遗传筛查 更快地确定基因和途径，改变产前乙醇暴露的易感性。这些 研究利用了具有不同酒精敏感性的各种小鼠品系， 转基因小鼠系，以及高通量斑马鱼遗传分析和CRISPR/Cas9基因编辑 技术.在目前的建议中，我们将进一步结合强大的 协作杂交小鼠遗传学工具的基因组分析能力。在目标1中，我们将探索 使用互补的小鼠和鱼转基因系，乙醇敏感性的潜在机制， 通过比较高度乙醇敏感的小鼠品系C57 BL/6 J， vs.高乙醇抗性菌株129 S1/Svlmj。这种比较将通过胚胎转录组学来辅助。 （RNA-Seq）分析，选择性杂交育种以诱导具有广泛抗性的抗性系（129）中的易感性， 基因组测序分析。Aim 2将通过检查CC创始人显著扩展我们的基因组分析 菌株对乙醇的敏感性，然后是敏感和抗性菌株的转录组学分析。 保守的候选基因和途径将进一步测试和特点，在我们的高通量 斑马鱼表型分析。目标3将采取补充生物信息学方法， 在高通量斑马鱼筛选中基因-乙醇相互作用的化学修饰剂，并进一步证实 在我们的FASD小鼠模型中。这项提案汇集了老鼠和鱼类遗传学、胚胎学 和酒精致畸学总之，这些实验将极大地增强我们对基因的理解。 胚胎发育早期乙醇诱导的脑和颅面畸形的病因，以及 有助于鉴定FASD的致病机制。

Project Summary/Abstract Alcohol (ethanol) exposure during pregnancy is the leading environmental cause of birth defects and central nervous system dysfunction. While the effects of ethanol on the brain and face have been explored quite extensively, there is considerable variation in the consequences of developmental ethanol exposure. Some of this variation is due to differences in timing and amount of exposure or nutritional factors. However, even when controlling for these factors, it is still clear that not everyone exposed to ethanol during development is affected equally and it has become increasingly clear that genetic factors play a very significant role in fetal alcohol spectrum disorders (FASD). Historically, elucidating these genetic factors that mediate risk or resilience has been relatively slow and characterized by human association studies or quantitative trait loci mapping in animal models. More recently, we have applied next generation sequencing technologies and forward genetic screens to more rapidly identify genes and pathways that alter susceptibility to prenatal ethanol exposure. These studies have taken advantage of varied mouse strains with differential alcohol susceptibility, numerous transgenic mouse lines, and high throughput zebrafish genetic analyses and CRISPR/Cas9 gene editing techniques. In this current proposal, we will further these approaches with the combination of the powerful genomic analyses capabilities of the Collaborative Cross mouse genetics tools. In Aim 1, we will explore mechanisms underlying ethanol sensitivity using complementary mouse and fish transgenic lines, while identifying further candidate genes via comparisons of the highly ethanol susceptible mouse strain, C57BL/6J vs. the highly ethanol resistant strain 129S1/Svlmj. This comparison will be aided by embryonic transcriptomic (RNA-Seq) analyses, selective crossbreeding to induce susceptibility in a resistant line (129) with extensive genome sequencing analyses. Aim 2 will significantly expand our genomic analyses by examining CC founder strains for their susceptibility to ethanol followed by transcriptomic profiling of susceptible and resistant strains. Conserved candidate genes and pathways will be further tested and characterized in our high throughput zebrafish phenotyping analyses. Aim 3 will take a complementary bioinformatic approach by identifying chemical modifiers of gene-ethanol interactions in a high throughput zebrafish screen with further confirmation in our mouse model of FASD. This proposal brings together experts on mouse and fish genetics, embryology and alcohol teratology. Together, these experiments will greatly enhance our understanding of the genetic etiology of ethanol-induced brain and craniofacial malformations during early embryonic development, as well as aiding in the identification of the pathogenic mechanisms involved in FASD.