Characterizing the Genetics of FASD in Complementary Mouse and Fish Models

项目概要/摘要 怀孕期间接触酒精（乙醇）是导致出生缺陷和中枢性缺陷的主要原因。 神经系统功能障碍。虽然乙醇对大脑和面部的影响已经被广泛探索 从广泛意义上讲，发育期乙醇暴露的后果存在相当大的差异。一些 这种差异是由于接触时间和数量或营养因素的差异造成的。然而，即使当 控制这些因素后，很明显并不是每个在开发过程中接触乙醇的人都会受到影响 同样，越来越明显的是，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复原力的遗传因素已经 相对缓慢，并以人类关联研究或动物数量性状基因座作图为特征 模型。最近，我们应用了下一代测序技术和正向遗传筛选 更快速地识别改变产前乙醇暴露易感性的基因和途径。这些 研究利用了具有不同酒精敏感性的不同小鼠品系，大量 转基因小鼠品系、高通量斑马鱼遗传分析和 CRISPR/Cas9 基因编辑 技术。在当前的提案中，我们将结合强大的技术来进一步推进这些方法 协作交叉小鼠遗传学工具的基因组分析功能。在目标 1 中，我们将探索 使用互补的小鼠和鱼转基因系的乙醇敏感性的机制，同时 通过比较高度乙醇敏感的小鼠品系 C57BL/6J 确定更多候选基因 与高度乙醇抗性菌株 129S1/Svlmj 相比。这种比较将得到胚胎转录组学的帮助 （RNA-Seq）分析，选择性杂交以诱导抗性品系（129）具有广泛的易感性 基因组测序分析。目标 2 将通过检查 CC 创始人来显着扩展我们的基因组分析 菌株对乙醇的敏感性，然后对敏感菌株和抗性菌株进行转录组分析。 保守的候选基因和通路将在我们的高通量中进一步测试和表征 斑马鱼表型分析。目标 3 将采取补充生物信息学方法，通过识别 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确认 在我们的 FASD 小鼠模型中。该提案汇集了小鼠和鱼类遗传学、胚胎学专家 和酒精畸胎学。总之，这些实验将极大地增强我们对遗传的理解。 早期胚胎发育过程中乙醇引起的脑部和颅面畸形的病因学，以及 有助于识别 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.