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

基本信息

批准号：
10663784
负责人：
THOMAS G FAZZIO
金额：
$ 51.59万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
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/10663784
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 Human Hybrids Individual 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 Shapes Signal Pathway Specific qualifier value Stem cell pluripotency TCF7L2 gene Testing Totipotency Totipotent Totipotent cell Yeasts apoAI regulatory protein-1 blastocyst cell fate specification developmental disease embryo stage 2 embryonic stem cell epigenomics gene regulatory network genome-wide genomic locus 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

项目摘要

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。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滋养层主调节因子（PMR）或滋养层主调节因子（TMR）仍然很大程度上未知。早期发展的认识至少是由于两个因素造成的：第一，技术障碍阻碍了使用。许多分子和基因组学方法都针对细胞数量较少的胚胎。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重大空白是必要的。我们提出了三个目标来解决早期胚胎基因调控网络（GRN）的问题。该问题的目标 1 重点是全面鉴定调节 PMR 的转录因子和 TMR 并揭示其调控功能，目标 2 将生成谱系解析的表观遗传学图谱。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在目标 3 中，我们建议剖析参与细胞命运规范的转录因子。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 模型来介导这一决定，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。