Characterization of the gene regulatory network governing the first cell fate decision in mammalian embryonic development

哺乳动物胚胎发育中第一个细胞命运决定的基因调控网络的表征

• 批准号: 10364821
• 负责人: THOMAS G FAZZIO
• 金额: $ 54.96万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-12 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10364821
• 关键词: Address; Architecture; Atlases; Binding; Binding Sites; Biological Assay; Cell Count; Cell Culture Techniques; Cell Line; Cells; ChIP-seq; Chromatin; Chromatin Structure; Coupled; Cultured Cells; DNA; DNA Methylation; Data Set; Development; Developmental Gene; Diagnosis; Embryo; Embryology; Embryonic Development; Epiblast; Epigenetic Process; Future; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genomic approach; Genomics; Human; Hybrids; Individual; Location; Maintenance; Mammalian Cell; Maps; Measures; Mediating; Methods; Modeling; Molecular; Molecular Genetics; Mus; Names; Nucleic Acid Regulatory Sequences; Nucleosomes; Pre-implantation Embryo Development; RNA Interference; Regenerative Medicine; Regulator Genes; Regulatory Element; Reporter; Role; Signal Pathway; Stem cell pluripotency; TCF7L2 gene; Testing; Totipotency; Totipotent; Totipotent cell; Yeasts; activating transcription factor; apoAI regulatory protein-1; base; blastocyst; cell fate specification; developmental disease; embryo stage 2; embryonic stem cell; epigenomics; gene regulatory network; genome-wide; histone modification; in vivo; in vivo evaluation; insight; novel strategies; pluripotency; prevent; regenerative approach; self-renewal; tool; transcription factor; transcriptome sequencing; transcriptomics; trophoblast; trophoblast stem cell

PROJECT SUMMARY Although the “master regulators” that maintain the self-renewal of pluripotent embryonic stem cells and extraembryonic trophoblast stem cells are relatively well understood, little is known about how these transcription factors are regulated in vivo at early stages of embryogenesis. Molecular genetics and embryology studies have identified signaling pathways required to direct cells to become trophoblast or embryonic epiblast, but the transcription factors that activate expression of pluripotency master regulators (PMRs) or trophoblast master regulators (TMRs) in each cell remain largely unknown. This major gap in our understanding of early development is due to at least two factors. First, technical barriers prevent the use of many molecular and genomics approaches in embryos consisting of low cell numbers. Second, there are significant discrepancies between the totipotent blastomeres of early embryos (where most PMRs and TMRs begin to be expressed) and cell culture models of totipotent cells. Therefore, a new approach that combines comprehensive methods for identification of regulators of PMR and TMR expression with sensitive new methods of testing their functions in vivo is necessary to fill this major gap in our understanding of the early embryonic gene regulatory network (GRN). We propose three aims to address this problem. Aim 1 is focused on comprehensive identification of transcription factors regulating PMRs and TMRs and uncovering their regulatory functions. Aim 2 will generate a lineage-resolved atlas of epigenetic changes that occur as cells select either the epiblast or trophoblast fate, as well as identify the direct targets of transcription factors that participate in cell fate specification. In Aim 3, we propose to dissect the mechanisms by which developmental transcription factors help elicit these epigenetic changes and generate a model of the totipotent GRN that mediates this decision. Successful completion of these studies will reshape our understanding of early embryonic gene regulation, as well as specification of epiblast and trophoblast cell fate.

项目概要 尽管维持多能胚胎干细胞自我更新的“主调节因子” 人们对胚胎外滋养层干细胞的了解相对较好，但对于这些细胞如何发挥作用却知之甚少。 转录因子在胚胎发生的早期阶段在体内受到调节。分子遗传学和 胚胎学研究已经确定了指导细胞成为滋养层或滋养层所需的信号通路 胚胎外胚层，但激活多能性主调节因子表达的转录因子 每个细胞中的 PMR（PMR）或滋养层主调节因子（TMR）仍然很大程度上未知。我们的这一重大差距 对早期发展的理解至少归因于两个因素。首先，技术壁垒阻碍了使用 许多分子和基因组学方法都适用于细胞数量较少的胚胎。其次，有 早期胚胎的全能卵裂球之间存在显着差异（其中大多数 PMR 和 TMR 开始表达）和全能细胞的细胞培养模型。因此，一种新方法 将鉴定 PMR 和 TMR 表达调节因子的综合方法与 测试其体内功能的敏感新方法对于填补我们的这一主要空白是必要的。 了解早期胚胎基因调控网络（GRN）。我们提出三个目标来解决 这个问题。目标 1 侧重于全面鉴定调节 PMR 的转录因子和 TMR 并揭示其监管功能。目标 2 将生成谱系解析的表观遗传图谱 当细胞选择外胚层或滋养层命运以及识别直接目标时发生的变化 参与细胞命运规范的转录因子。在目标 3 中，我们建议剖析 发育转录因子帮助引发这些表观遗传变化的机制 生成一个全能 GRN 模型来调节这一决策。成功完成这些研究 将重塑我们对早期胚胎基因调控以及外胚层和外胚层规范的理解 滋养层细胞的命运。