Novel Roles of Pluripotency Regulators in the Early Mouse Embryo

多能性调节因子在早期小鼠胚胎中的新作用

• 批准号: 7798511
• 负责人: Wing H. WONG
• 金额: $ 33.32万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-07 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7798511
• 关键词: Address; Architecture; Boxing; Cell Cycle; Cell Lineage; Cells; Competence; Data; Development; Embryo; Embryonic Development; Ensure; Epigenetic Process; Family; Fertilization; Fetus; Figs - dietary; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Programming; Genome; Germ Cells; Goals; Human; In Vitro; Infertility; Inner Cell Mass; Lead; Malignant Neoplasms; Medicine; Modeling; Molecular Profiling; Morula; Mus; POU domain factors; Phenotype; Regulation; Regulator Genes; Reporting; Role; Signal Pathway; Signaling Molecule; Staging; Stem Cell Research; Stem cells; Testing; Transcript; Translations; Up-Regulation; Work; anticancer research; base; blastocyst; cyclin A2; embryo stage 2; embryonic stem cell; gene discovery; gene function; homeodomain; human embryonic stem cell; member; novel; nuclear reprogramming; pluripotency; programs; public health relevance; regenerative; self-renewal; somatic cell nuclear transfer; zygote

DESCRIPTION (provided by applicant): The gene regulatory network that controls pluripotency and differentiation in embryonic stem cells (ESCs) is widely studied and becoming well understood. However, the genetic parameters of successful somatic cell nuclear transfer and the genetic program that sets up the ESC gene network are not known. The answers to these key questions in ESC research lie in the gene regulation of the early embryo, which for our purpose here, encompasses developmental stages that occur after fertilization and prior to blastocyst formation. Just as the inner cell mass of the blastocyst must maintain pluripotency and the ability to self-renew in order to give rise to cell lineages that form the fetus, the blastomeres of the early embryo must establish totipotency and self-renewal abilities to form the blastocyst. However, we do not know how totipotency of blastomeres is achieved and how it relates to the waves of embryonic genome activation. Therefore, our overall goal is to address both the general architecture of this gene network and identify specific regulators that are critical and sufficient for establishing the correct genetic circuitry at the 1- to 2-cell stages, and that would ensure subsequent developmental competence. Further, the function of these regulators will be determined in order to understand mechanisms that are essential in early embryo development. We have established experimental strategies to interrogate the precise roles of transcriptional regulators during the maternal-embryonic transition of the early embryo, when both maternal and early embryonic transcripts may be present simultaneously. We discovered that Oct4, a homeodomain transcription factor of the POU family that is known for its critical functions in pluripotency in the inner cell mass, ESCs, and germ cells, has a novel role in early embryo development prior to the blastocyst stage and is required for progression beyond the multi-cell and morula stages. In addition, our data suggest that the pluripotency regulators, Sox2 and Sall4, may also have critical functions prior to blastocyst formation. We propose to fully investigate the novel roles of these and other pluripotency regulators and use them as "portals" to dissect gene regulation in the early embryo in the context of nuclear reprogramming and embryonic genome activation. Understanding the genetic requirements of the early embryo will have a direct and significant impact on regenerative and stem cell medicine, the treatment of infertility, and cancer research, as genes implicated in cancer are highly enriched in this genetic program. Public Health Relevance: The overall goal of this project is to investigate the novel roles of Oct4, Sall4, Sox2, and other pluripotency regulators in the early mouse embryo, with a focus on the 1- to 2-cell stages during the maternal-embryonic transition. Using these transcriptional regulators as "portals", we will dissect the gene regulatory network in the early embryo and identify specific regulators that are critical and sufficient for ensuring subsequent developmental competence. Understanding the genetic requirements of the early embryo will have a direct and significant impact on regenerative and stem cell medicine, the treatment of infertility, and cancer research.

描述（由申请人提供）：控制胚胎干细胞中多能性和分化（ESC）的基因调节网络（ESC）已被广泛研究并逐渐理解。但是，尚不清楚成功的体细胞核转移的遗传参数和建立ESC基因网络的遗传程序。 ESC研究中这些关键问题的答案在于早期胚胎的基因调节，出于我们的目的，它涵盖了受精后和胚泡形成之前发生的发育阶段。正如胚泡的内部细胞质量必须保持多能性和自我更新的能力，以产生形成胎儿的细胞谱系一样，早期胚胎的囊泡必须建立甲壳虫和自我更新能力以形成胚泡。但是，我们不知道如何实现胚泡的全能性以及它与胚胎基因组激活的波浪的关系。因此，我们的总体目标是解决该基因网络的一般体系结构，并确定特定的调节剂，这些调节器至关重要，足以在1至2个细胞阶段建立正确的遗传电路，这将确保后续发展能力。此外，将确定这些调节剂的功能，以了解早期胚胎发育中必不可少的机制。我们已经建立了实验策略，以询问早期胚胎的母体 - 胚胎过渡期间转录调节剂的精确作用，当时母体和早期的胚胎转录本可以同时存在。我们发现，POU家族的同源域的转录因子Oct4以其在内部细胞质量，ESC和生殖细胞中多能在多能性方面的关键功能而闻名，在胚泡阶段之前的早期胚胎发育中具有新颖的作用，并且是多细胞和害怕阶段以外的胚胎发展所必需的。此外，我们的数据表明，多能调节器SOX2和SALL4在胚泡形成之前也可能具有关键功能。我们建议在核重编程和胚胎基因组激活的背景下充分研究这些和其他多能调节剂的新作用，并将其用作早期胚胎中基因调节的“门户”。了解早期胚胎的遗传需求将对再生和干细胞医学，不育治疗以及癌症研究产生直接而重大的影响，因为与癌症有关的基因在此遗传程序中高度丰富。 公共卫生相关性：该项目的总体目标是调查Oct4，Sall4，Sox2和其他多能调节器在早期小鼠胚胎中的新作用，重点是在母体 - 胚胎过渡期间的1至2个细胞阶段。将这些转录调节剂作为“门户”，我们将在早期胚胎中剖析基因调节网络，并确定特定的调节剂，这些调节剂至关重要，足以确保后续的发展能力。了解早期胚胎的遗传要求将对再生和干细胞医学，不育治疗以及癌症研究产生直接而重大的影响。