Characterizing the Genetics of FASD in Complementary Mouse and Fish Models

在互补小鼠和鱼类模型中描述 FASD 的遗传学特征

• 批准号: 10792720
• 负责人: JOHANN K EBERHART
• 金额: $ 56.69万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10792720
• 关键词: Affect; Alcohols; Animal Model; Bioinformatics; Biological Assay; Brain; C57BL/6N Mouse; CRISPR/Cas technology; Candidate Disease Gene; Cell Line; Central Nervous System; Chemical Modifier; Chemical-Induced Change; Chemicals; Collaborations; Congenital Abnormality; Craniofacial Abnormalities; Crossbreeding; Data; Development; Embryo; Embryology; Embryonic Development; Ethanol; Face; Fetal Alcohol Exposure; Fetal Alcohol Spectrum Disorder; Fishes; Functional disorder; Gene Expression; Gene Modified; Genes; Genetic; Genetic Predisposition to Disease; Genetic Screening; Genomics; Goals; Human; Individual; Intervention; Knowledge; Laboratories; Maps; Mediating; Modeling; Mouse Strains; Mus; Mutation; Nature; Nutritional; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Phenotype; Play; Predisposition; Pregnancy; Prevention; Quantitative Trait Loci; Rapid screening; Reader; Resistance; Resources; Risk; Role; Techniques; Technology; Teratogens; Teratology; Testing; Tissue-Specific Gene Expression; Transcription Alteration; Transgenic Mice; Transgenic Organisms; Variant; Work; Zebrafish; alcohol effect; alcohol exposure; alcohol sensitivity; brain abnormalities; brain malformation; candidate identification; differential expression; experimental study; gene conservation; genetic analysis; genome sequencing; improved; insight; interest; mouse genetics; mouse model; mutant; next generation sequencing; novel; resilience; resistant strain; teratogenesis; tool; transcriptome; transcriptome sequencing; transcriptomic profiling; transcriptomics

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.

项目摘要/摘要 怀孕期间酒精（乙醇）暴露是出生缺陷的主要环境原因 神经系统功能障碍。虽然已经探索了乙醇对大脑和脸部的影响 广泛地，发育性乙醇暴露的后果存在很大差异。其中一些 这种差异是由于时间的计时和暴露量或营养因素的差异所致。但是，即使在那时 控制这些因素，很明显，在开发过程中并不是每个人都暴露于乙醇 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 频谱障碍（FASD）。从历史上看，阐明了介导风险或韧性具有的这些遗传因素 相对较慢，以人类关联研究或动物的定量性状基因映射为特征 型号。最近，我们应用了下一代测序技术和前进遗传筛选 更快地识别改变对产前乙醇暴露易感性的基因和途径。这些 研究利用了具有差异性酒精敏感性的各种小鼠菌株，许多 转基因小鼠系和高吞吐量斑马遗传分析和CRISPR/CAS9基因编辑 技术。在当前的建议中，我们将通过强大的结合进一步进一步 基因组分析协作跨小鼠遗传学工具的功能。在AIM 1中，我们将探索 使用互补小鼠和鱼类转基因线的乙醇敏感性的机制，而 通过比较高度乙醇易感小鼠菌株C57BL/6J来鉴定进一步的候选基因 与高度乙醇抗性菌株129S1/SVLMJ。该比较将通过胚胎转录组来帮助 （RNA-seq）分析，选择性杂交以在耐药线（129）中引起易感性（129） 基因组测序分析。 AIM 2将通过检查CC创始人大大扩展我们的基因组分析 其对乙醇的敏感性，然后是易感和抗性菌株的转录组分析。 保守的候选基因和途径将在我们的高通量中进一步测试和表征 斑马鱼表型分析。 AIM 3将通过识别来采用互补的生物信息学方法 在高吞吐量斑马鱼筛查中的基因 - 乙醇相互作用的化学修饰符，并进一步确认 在我们的FASD鼠标模型中。该建议汇集了有关鼠标和鱼类遗传学的专家 和酒精畸形学。这些实验将大大增强我们对遗传的理解 早期胚胎发育过程中乙醇引起的大脑和颅面畸形的病因 作为鉴定FASD涉及的致病机制的鉴定。