Ethanol-induced epigenetic changes in the early embryo

胎儿酒精谱系障碍(FASD)是由产前酒精暴露引起的颅面部 畸形、认知障碍、感觉障碍、运动障碍和器官畸形。FASD 在美国人口中出生的儿童中有3%-10%会发生这种情况。该项目的长期目标是了解 FASD的分子机制和确定治疗靶点。总体假设是乙醇 暴露会产生有害的表观遗传和基因表达效应，从而破坏关键的早期 胚胎发生事件，可通过叶酸(FA)处理挽救。早期发育(原肠形成)是一种 酒精引起的缺陷的高度敏感期，在人类中，原肠形成发生在植入时，第一次 可能接触母体血液循环的时间，从而接触母体血液酒精的时间。发育基因 表达和表观遗传事件在脊椎动物中高度保守。我们的数据指向乙醇和FA- 早期斑马鱼胚胎中发生的敏感原肠形成事件。FA是单碳代谢的一部分 促进DNA和组蛋白的甲基加成的途径。了解FA的保护作用将 帮助我们更好地针对出生缺陷预防策略，并潜在地确定新的治疗策略。我们的 初步的基因表达研究表明Sox2和Elf3转录因子对乙醇敏感 在早期胚胎中暴露，是广泛转录网络的一部分。具体目标1：确定 全基因组DNA甲基化、染色质结构和基因表达的变化 酒精和甲醛暴露。拟议的实验将分析全基因组乙醇和FA对DNA的影响- 甲基化(MeDIP-seq)、染色质结构(atac-seq)和基因表达数据(rna-seq) 平行的。该数据集将确定乙醇是否影响表观遗传事件、基因表达以及 FA可以预防这些表观遗传缺陷。质询组蛋白在特定基因启动子上的修饰 靶向染色质免疫沉淀和定量聚合酶链式反应(芯片-聚合酶链式反应)将分析表观遗传机制 发生在大的染色质结构变化期间(ATAC-seq)。具体目标2：分析潜在基因 乙醇诱导胚胎发育缺陷发生过程中sox2和elf3的相互作用。建议 实验将评估sox2和elf3的单独作用以及sox2和elf3之间的潜在相互作用。 并确定这些相互作用在我们的FASD中是否具有功能意义 模特。这个整体项目拥有关于乙醇在表观遗传事件中的作用的可靠答案的专业知识， 基因表达程序和功能结果。乙醇在早期的作用机制 发育是一个知识鸿沟，这项研究将在检测基因的同时考察表观遗传效应 表情效果。未来的治疗方法将取决于我们对出生缺陷综合征的了解 创世纪。

Fetal alcohol spectrum disorder (FASD) is caused by prenatal alcohol exposure, resulting in craniofacial dysmorphology, cognitive impairment, sensory disabilities, motor disabilities and organ deformities. FASD occurs in 3-10% of children born in US populations. The long-term goal of this project is to understand the molecular mechanisms and to identify therapeutic targets for FASD. The overall hypothesis is that ethanol exposure produces detrimental epigenetic and gene expression effects that disrupt critical early embryogenesis events, which are rescued by folic acid (FA) treatment. Early development (gastrulation) is a highly sensitive period for ethanol-induced defects, and in humans, gastrulation occurs at implantation, the first possible time to be exposed to maternal circulation and thus, maternal blood alcohol. Developmental gene expression and epigenetic events are highly conserved in vertebrates. Our data point to ethanol and FA- sensitive gastrulation events occurring in early zebrafish embryos. FA is part of one-carbon metabolism pathways that facilitate methyl group addition to DNA and histones. Understanding FA protective effects will help us better target birth defect prevention strategies and potentially identify new therapeutic strategies. Our preliminary gene expression studies showed that Sox2 and Elf3 transcription factors are sensitive to ethanol exposure in the early embryo and are parts of an extensive transcriptional network. Specific aim 1: Determine genome-wide changes in DNA-methylation, chromatin structure and gene expression in response to early ethanol and FA exposure. Proposed experiments will analyze genome-wide ethanol and FA effects on DNA- methylation (MeDIP-seq), chromatin structure (ATAC-seq) and gene expression data (RNA-seq) collected in parallel. This dataset will determine whether ethanol affects epigenetic events, gene expression and whether FA can prevent these epigenetic defects. Interrogating histone modifications at specific gene promoters by targeted chromatin immunoprecipitation and quantitative PCR (ChIP-PCR) will analyze epigenetic mechanisms occurring during large chromatin structural changes (ATAC-seq). Specific aim 2: Analyze potential gene interactions between sox2 and elf3 during ethanol-induced embryogenesis defect genesis. Proposed experiments will evaluate individual roles of sox2 and elf3 and potential interactions between sox2 and elf3 during early development and determine whether these interactions have functional significance in our FASD model. This overall project has the expertise to get robust answers about ethanol's role in epigenetic events, gene expression programs and functional consequences. The mechanism of action for ethanol during early development is a knowledge gap, and this study will examine epigenetic effects in parallel with gene expression effects. Future therapeutic approaches will rely on our knowledge of the birth defect syndrome's genesis.