Epigenetic and environmental impact on early embryonic development

表观遗传和环境对早期胚胎发育的影响

• 批准号: 10242497
• 负责人: Zachary Dylan Smith
• 金额: $ 150.75万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10242497
• 关键词: Accounting; Address; Affect; Cell Line; Cells; Chromatin; Code; Complex; Congenital Abnormality; Congenital Disorders; DNA; Data; Development; Developmental Process; Diagnostic; Dimensions; Embryo; Embryonic Development; Environment; Environmental Impact; Enzymes; Epigenetic Process; Fetal Diseases; Folic Acid; Foundations; Gene Expression Regulation; Generations; Genes; Genetic; Genome; Health; Human; Intervention; Methods; Micromanipulation; Modeling; Molecular; Morphology; Nature; Organism; Organogenesis; Pathology; Pathway interactions; Penetrance; Phenotype; Polycomb; Post-Translational Protein Processing; Pregnancy; Process; Regulation; Repression; Reproducibility; Reproductive Medicine; Resolution; Series; Specific qualifier value; Specificity; System; Techniques; Tissues; Transgenic Organisms; Validation; Variant; cell type; chromatin remodeling; cohort; design; dietary; epigenetic regulation; fetal; flexibility; indexing; innovation; mutant; new technology; novel; programs; reproductive; single cell analysis; tool; transcription factor

Project Summary/Abstract: During development, cellular diversity is achieved via the dynamic interpretation of a fixed DNA template, a process that incorporates both straightforward genetic and obscure epigenetic concepts. Cell identity is in part governed by transcription factor-directed programs, but these interact with elaborate “codes” of post-translational modifications to chromatin that index regions of the genome for activity or repression. To date, the general relationships between local chromatin remodeling and gene regulation have been primarily modeled in cell lines and cannot yet account for the intricacy of the developing embryo. Moreover, although many epigenetic regulators are essential for viable embryogenesis, it remains unclear how these enzymes participate in specific developmental processes given the generic nature of their target substrates. A detailed accounting of how epigenetic regulation operates during gestation is critical to understand complex congenital or fetal disorders, which often have unclear penetrance, affect multiple cell types or tissue systems, and can be highly influenced by the maternal environment. Here, I propose a transformative new strategy for comprehensively phenotyping mutant embryos that incorporates detailed micromanipulation techniques, novel molecular systems, and single cell analysis to recover high resolution morphological, molecular and temporal information from many replicates simultaneously. My flexible approach permits rapid transition from hypothesis to validation and eliminates many cumbersome aspects of traditional transgenics, as well as the frequent limitation of examining only a limited number of lineages via a handful of pre-specified marker genes. I will apply this pipeline to dissect the specific regulatory impact of maternal dietary folate and of the Polycomb group repressors on early organogenesis, which represent “top-down” and “bottom-up” approaches to interpret developmental robustness, the deliberate and reproducible generation of a sophisticated organism under uncertain and fluctuating conditions. To do so, I will leverage my ability to recover data from mutant cohorts and optimize new analytical strategies to quantify phenotypic variation and incomplete penetrance. These factors are central concerns of reproductive medicine and have been extraordinarily challenging to reduce to clear molecular mechanisms or pathways, subsequently limiting innovation of novel interventions and diagnostics. In addition to addressing fundamental human health issues, these efforts will also substantially advance a suite of new tools for exogenously titrating specific regulatory components as well as for recording the historical relationship between single cells. If successful, this proposal will uncover previously opaque dimensions of embryonic regulation, including shared and cell-type specific consequences of epigenetic or environmental insults, as well as transform general methods for characterizing previously intractable congenital abnormalities.

项目概要/摘要：在开发过程中，通过动态解释实现细胞多样性 一个固定的DNA模板，一个过程，结合了简单的遗传和模糊的表观遗传 理念的细胞身份部分由转录因子指导的程序控制，但这些程序与转录因子相互作用。 对染色质的翻译后修饰的精细“代码”，其指示基因组区域的活性，或 镇压到目前为止，局部染色质重塑和基因调控之间的一般关系， 主要是在细胞系中建模，还不能解释发育中胚胎的复杂性。此外，委员会认为， 尽管许多表观遗传调节因子对有活力的胚胎发生是必需的，但仍不清楚这些调节因子是如何作用的。 酶参与特定的发育过程，因为它们的靶底物具有一般的性质。一 详细说明怀孕期间表观遗传调控是如何运作的，对于理解复杂的 先天性或胎儿疾病通常具有不清楚的遗传性，影响多种细胞类型或组织系统， 并且很容易受到母体环境的影响。在这里，我提出了一个变革性的新战略， 全面表型突变胚胎，结合详细的显微操作技术，新的 分子系统和单细胞分析，以恢复高分辨率的形态，分子和时间 同时从多个副本中获取信息。我灵活的方法允许从假设快速过渡 验证和消除了许多繁琐的方面，传统的转基因，以及频繁的 通过少数预先指定的标记基因仅检查有限数量的谱系的局限性。我会申请 这条管道剖析了母体膳食叶酸和Polycomb组的特定调节影响， 阻遏物对早期器官发生的影响，这代表了“自上而下”和“自下而上”的方法来解释 发育的健壮性，一个复杂的有机体在 不确定和波动的条件。为此，我将利用我的能力从变种人群体中恢复数据， 优化新的分析策略，以量化表型变异和不完全突变。这些因素 是生殖医学的核心问题，并且非常具有挑战性， 分子机制或途径，从而限制了新干预和诊断的创新。在 除了解决基本的人类健康问题外，这些努力还将大大推动一系列 新的工具，用于外源滴定特定的监管成分，以及记录历史 单细胞之间的关系。如果成功，这项提议将揭示以前不透明的层面， 胚胎调节，包括表观遗传或环境的共同和细胞类型特定后果 侮辱，以及改造的一般方法来表征以前棘手的先天性异常。