Role of ETS-Transcription Factors in Regulating Cell Fate

ETS 转录因子在调节细胞命运中的作用

• 批准号: 10486990
• 负责人: Sergio Ruiz
• 金额: $ 35.26万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10486990
• 关键词: Binding; Cell Nucleus; Cell Proliferation; Cells; Data; Defect; Development; Embryonic Development; Enhancers; Ensure; Extinction (Psychology); Family; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Goals; Growth; KRAS2 gene; MAP Kinase Gene; Malignant Neoplasms; Membrane; Methyltransferase; Mus; NRAS gene; Nuclear; Proteins; Publications; RAS genes; Receptor Signaling; Role; Science; Transcription Repressor; Work; embryonic stem cell; pluripotency; programs; ras Proteins; transcription factor; transcriptome sequencing

During FY2021 we have made great progress in this project, and we have this work accepted for publication in Sciences Advances. We have established the role of the transcriptional repressor ERF as the MAPK dependent switch that controls the exit from the naive state of pluripotency. For this, we have used inducible ESCs to genetically eliminate all RAS proteins and determined that, while differentiated RAS deficient cells remain trapped in an intermediate state of pluripotency, further absence of ERF overcomes the developmental blockage of RAS deficient cells from this intermediate state. We also performed RNAseq data and observed that deletion of ERF restored the overall gene expression profile to a wild type level in differentiated Ras deficient cells. Mechanistically, we have demonstrated that ERF ensures naive pluripotency by strengthen naive pluripotent transcription factor binding and accessibility at specific ESC enhancers. Moreover, we have also determined that ERF regulates negatively the expression of the de novo methylase DNMT3, essential for the extinction of the naive transcriptional program. Our data has revealed an essential role for ERF in the exit from rosette pluripotency as a regulator of the progression to primed pluripotency.

2021 财年，我们在该项目上取得了巨大进展，这项工作已被《Science Advances》接受发表。我们已经确定了转录抑制子 ERF 作为 MAPK 依赖性开关的作用，控制着多能性幼稚状态的退出。为此，我们使用诱导型ESC从基因上消除所有RAS蛋白，并确定，虽然分化的RAS缺陷细胞仍被困在多能性的中间状态，但ERF的进一步缺失克服了RAS缺陷细胞从该中间状态的发育障碍。我们还进行了 RNAseq 数据，并观察到 ​​ERF 的删除将分化的 Ras 缺陷细胞中的整体基因表达谱恢复到野生型水平。从机制上讲，我们已经证明 ERF 通过加强幼稚多能转录因子结合和特定 ESC 增强子的可及性来确保幼稚多能性。此外，我们还确定 ERF 负向调节从头甲基化酶 DNMT3 的表达，这对于幼稚转录程序的灭绝至关重要。我们的数据揭示了 ERF 在玫瑰花结多能性退出中的重要作用，作为启动多能性进展的调节因子。