Transcriptional and Epigenetic Control of Pluripotency and Development by Zfp281

Zfp281 对多能性和发育的转录和表观遗传控制

基本信息

批准号：
9903392
负责人：
Jianlong Wang
金额：
$ 32.4万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-06 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9903392
关键词：
Alleles Cell model DNA DNMT3a Development Disease Embryo Embryonic Development Enzymes Epiblast Epigenetic Process Event Genes Genetic Transcription Genomics Goals Human In Vitro Inner Cell Mass Knock-out Knockout Mice Knowledge Maintenance Malignant Neoplasms Mammalian Cell Methylation Mixed Function Oxygenases Modeling Molecular Mouse Strains Mus Nodal Play Pluripotent Stem Cells Proteins Proteomics Role Signal Transduction Somatic Cell Stem Cell Development System Testing Tissues Transcription Coactivator Transcription Process Work blastocyst cofactor conditional knockout differential expression embryonic stem cell epigenome human embryonic stem cell implantation in vivo induced pluripotent stem cell insight mouse model natural Blastocyst Implantation novel pluripotency preimplantation recruit screening stem cells transcription factor

项目摘要

PROJECT SUMMARY Pluripotency is controlled by key transcription factors such as Nanog, Oct4, and Sox2 in conjunction with many epigenetic regulators including DNA (de)methylation enzymes, which together form the interconnected pluripotency regulatory networks. Mouse pluripotent stem cells exist in two distinct stable pluripotent states (naive and primed), which are represented by ground state embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs), respectively. Human ESCs share defining features of primed pluripotency with mouse EpiSCs, and naive human ESCs/iPSCs can also be established, although our understanding of naive and primed pluripotency of hESCs is very limited. In pursuit of a deeper understanding of pluripotency under defined naïve and primed culture conditions, we have recently discovered Zfp281 and Tet1/2 as critical players for controlling pluripotent states in vitro. While in vitro studies are instrumental, it remains critical to confirm whether the insights they provide are relevant to events taking place in vivo during epiblast maturation from naive pre- implantation blastocyst to the primed post-implantation epiblast. We will employ our integrated genomics and proteomics approach combined with in vivo mouse models to dig deeper into mechanism in this application with a broader effort to dissect novel regulatory mechanism by which Zfp281/ZNF281 orchestrates transcriptional and epigenetic processes in controlling primed versus naive pluripotency in both in vitro cellular model and in vivo mouse knockout models. We will test the hypothesis that the pluripotency factor Zfp281/ZNF281 plays critical roles in coordinating opposing functions of Tet1 and Tet2 and the crosstalk between Tet1/2 and DNMT3a/3b proteins for transcriptional and epigenetic control of pluripotent states in both mouse and human systems. Our proposed studies encompass the following three Specific Aims. 1) Define the functional connection between Zfp281-DNMTs in controlling pluripotent states. 2) Define the functional connection between Zfp281 and TETs for primed pluripotency. 3) Investigate transcriptional and epigenetic regulatory roles of Zfp281 in epiblast maturation of mouse early embryos. Our work represents a conceptual advance and a fundamental contribution to the understanding of early mammalian development, which is grounded by pluripotent state transitions. Our combined approach to study Zfp281 with both cellular and mouse models of pluripotent state transitions will provide fundamental knowledge on finely coordinated transcriptional and epigenetic control of mammalian cell fate changes and embryonic development, dysregulation of which is often associated with disease and cancer.

项目概要多能性由关键转录因子（例如 Nanog、Oct4 和 Sox2）以及许多转录因子共同控制。表观遗传调节因子，包括 DNA（去）甲基化酶，它们共同形成了相互关联的小鼠多能干细胞存在两种不同的稳定多能状态。（初始的和引发的），以基态胚胎干细胞 (ESC) 和外胚层干细胞为代表细胞（EpiSC），分别与小鼠 EpiSC 共享启动多能性的定义特征。也可以建立初始的人类 ESC/iPSC，尽管我们对初始和启动的理解 hESC 的多能性非常有限。和引发的培养条件，我们最近发现 Zfp281 和 Tet1/2 作为控制的关键参与者虽然体外研究很有帮助，但确认是否具有多能状态仍然至关重要。他们提供的见解与幼稚前胚层成熟过程中体内发生的事件相关。我们将利用我们的整合基因组学和蛋白质组学方法与体内小鼠模型相结合，可更深入地挖掘该应用中的机制更广泛地努力剖析 Zfp281/ZNF281 协调的新调控机制转录和表观遗传过程在控制体外细胞的启动多能性与幼稚多能性中的作用我们将测试多能性因子的假设。 Zfp281/ZNF281 在协调 Tet1 和 Tet2 的相反功能以及串扰方面发挥着关键作用 Tet1/2 和 DNMT3a/3b 蛋白之间的多能状态转录和表观遗传控制我们提出的研究包括以下三个具体目标 1) 定义。 Zfp281-DNMT 之间控制多能状态的功能连接 2) 定义功能。 Zfp281 和 TET 之间的连接以实现引发的多能性 3) 研究转录和表观遗传。 Zfp281 在小鼠早期胚胎外胚层成熟中的调节作用我们的工作代表了一个概念。对增进对早期哺乳动物发育的了解做出了根本性贡献我们以多能状态转换为基础研究 Zfp281。多能状态转换的小鼠模型将提供精细协调的基础知识哺乳动物细胞命运变化和胚胎发育的转录和表观遗传控制，其失调通常与疾病和癌症有关。