Pluripotency transcription factor function during primordial germ cell development

原始生殖细胞发育过程中的多能转录因子功能

• 批准号: BB/L002736/1
• 负责人: Ian Chambers
• 金额: $ 58.79万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL002736%2F1
• 关键词: Pluripotency; transcription; factor; function; during

The aim of the proposed work is to study the factors regulating formation of functional germ cells, oocytes in females and sperm in males. We will focus on the role of Nanog, an important molecule involved at distinct stages during embryonic development.We will determine what parts of Nanog are required to mediate function in developing primordial germ cells (PGCs for short), the embryonic cells that will give rise to the adult germ cells. We will compare Nanog proteins from different species for their ability to function in mouse cells, since the mouse is the mammalian species where these processes are best understood. We will investigate whether Nanog function is unique, or whether other factors, such as Esrrb, can play a similar role in PGCs. We will determine how Nanog exerts its function in driving germ cell development by identifying the genes regulated by Nanog in this process.This study will delineate the mechanisms through which Nanog functions. We will learn if Nanog function has been conserved throughout evolution and which parts of the Nanog molecule are important. In a broader sense, since primordial germ cells and the cells of the early embryo that retain unrestricted differentiation potential are similar, we will understand better how the factors required for the formation of these distinct cells work. Understanding how functional germ cells are formed is of great importance for the progress of reproductive medicine. Learning how pluripotent cells are regulated has high impact on regenerative medicine strategies. The continued existence of a sexually reproducing species requires effective formation of germ cells that possess a haploid DNA complement and that can join with another germ cell of the opposite sex during fertilisation to form a new individual. PGC development from cells set aside early in development involves changes in gene expression and modifications of the chromatin, the complex of DNA and protein in which the genome is packed, as the PGCs migrate to the site where the genitals form, known as the genital ridges. Once the PGCs enter the genital ridges they undergo further changes to their chromatin that erase memories of their parental origin. This sets a blank slate onto which instructions for gene expression in the next generation can be written. These changes are directed by a number of important factors able to influence the expression of other genes, amongst which is Nanog. We have shown that Nanog is required for continued PGC development during the period of chromatin remodelling. Interestingly, Nanog is also required at the other developmental period in which profound chromatin remodelling occurs; the formation of cells with unrestricted differentiation potential (pluripotency) in the pre-implantation embryo. Pluripotency can be conveniently studied in cells grown in culture and derived from pre-implantation embryos: embryonic stem (ES) cells. Nanog controls the ability of ES cells to indefinitely duplicate. Our recent work has characterized the genes acting downstream of Nanog in ES cells. We consistently found that the gene most up-regulated by Nanog induction was Esrrb. We further showed that Esrrb could recapitulate many of the functional properties previously ascribed to Nanog and, importantly could do this in the absence of Nanog. A further key finding is that, in ES cells lacking Esrrb, the ability of Nanog to sustain pluripotency is compromised. This demonstrates unequivocally that Esrrb is a critical downstream mediator of Nanog function in ES cells.We now propose to investigate in greater detail the importance of Nanog function in germ cell development and determine whether a similar functional relationship exists between Nanog and Esrrb in PGCs. Our study has the potential to highlight new similarities between the pluripotent cells of the pre-implantation embryo and PGCs, ultimately improving our understanding of the principles governing both cell populations.

本研究的目的是研究调节功能性生殖细胞、雌性卵母细胞和雄性精子形成的因素。我们将重点关注Nanog的作用，这是一个在胚胎发育的不同阶段参与的重要分子。我们将确定Nanog的哪些部分需要介导发育原始生殖细胞（PGCs）的功能，这些胚胎细胞将产生成人生殖细胞。我们将比较来自不同物种的Nanog蛋白在小鼠细胞中的功能，因为小鼠是最了解这些过程的哺乳动物物种。我们将研究Nanog的功能是否独特，或者其他因素，如Esrrb，是否在PGCs中发挥类似的作用。我们将通过鉴定在这一过程中受Nanog调控的基因来确定Nanog在驱动生殖细胞发育中如何发挥其功能。本研究将描述Nanog的作用机制。我们将了解Nanog的功能是否在整个进化过程中一直保持不变，以及Nanog分子的哪些部分是重要的。从更广泛的意义上说，由于原始生殖细胞和早期胚胎的细胞具有不受限制的分化潜力是相似的，我们将更好地了解形成这些不同细胞所需的因素是如何工作的。了解功能性生殖细胞是如何形成的，对生殖医学的发展具有重要意义。了解多能性细胞是如何被调控的对再生医学策略有很大的影响。有性生殖物种的持续存在需要有效的生殖细胞形成，这些生殖细胞具有单倍体DNA补体，并且可以在受精过程中与另一个异性生殖细胞结合形成新的个体。在发育早期被搁置的细胞中，PGC的发育涉及基因表达的变化和染色质的修饰，染色质是DNA和蛋白质的复合体，基因组被包裹在染色质中，随着PGC迁移到生殖器形成的位置，即生殖器脊。一旦PGCs进入生殖器脊，它们的染色质就会发生进一步的变化，从而抹去它们父母起源的记忆。这为下一代基因表达的指令奠定了一个空白的基础。这些变化是由一些能够影响其他基因表达的重要因素指导的，其中包括Nanog。我们已经证明，在染色质重塑期间，Nanog是PGC持续发展所必需的。有趣的是，在染色质重塑发生的其他发育时期也需要Nanog；着床前胚胎中具有无限制分化潜能（多能性）的细胞的形成。多能性可以方便地在培养细胞和植入前胚胎中获得的细胞：胚胎干细胞（ES）中研究。Nanog控制胚胎干细胞无限复制的能力。我们最近的工作描述了在胚胎干细胞中作用于Nanog下游的基因。我们一致发现Nanog诱导上调最多的基因是Esrrb。我们进一步表明，Esrrb可以重现以前归因于Nanog的许多功能特性，重要的是，它可以在没有Nanog的情况下做到这一点。进一步的关键发现是，在缺乏Esrrb的胚胎干细胞中，Nanog维持多能性的能力受到损害。这明确表明Esrrb是胚胎干细胞中Nanog功能的关键下游介质。我们现在建议更详细地研究Nanog功能在生殖细胞发育中的重要性，并确定Nanog和Esrrb在PGCs中是否存在类似的功能关系。我们的研究有可能突出着床前胚胎的多能细胞和PGCs之间的新相似性，最终提高我们对控制这两种细胞群的原理的理解。

项目成果

Differential repression of Otx2 underlies the capacity of NANOG and ESRRB to induce germline entry

Otx2 的差异抑制是 NANOG 和 ESRRB 诱导种系进入能力的基础

• DOI: 10.1101/2021.06.14.448276
• 发表时间: 2021
• 作者: Vojtek M

Distinct Contributions of Tryptophan Residues within the Dimerization Domain to Nanog Function.

• DOI: 10.1016/j.jmb.2016.12.001
• 发表时间: 2017-05-19
• 期刊: Journal of molecular biology
• 影响因子: 5.6
• 作者: Mullin NP;Gagliardi A;Khoa LTP;Colby D;Hall-Ponsele E;Rowe AJ;Chambers I
• 通讯作者: Chambers I

OTX2 restricts entry to the mouse germline.

OTX2限制了进入鼠标种系的输入。

• DOI: 10.1038/s41586-018-0581-5
• 发表时间: 2018-10
• 期刊: Nature
• 影响因子: 64.8
• 作者: Zhang J;Zhang M;Acampora D;Vojtek M;Yuan D;Simeone A;Chambers I
• 通讯作者: Chambers I

Esrrb Complementation Rescues Development of Nanog-Null Germ Cells.

ESRRB互补挽救了纳米无效生殖细胞的发展。

• DOI: 10.1016/j.celrep.2017.12.060
• 发表时间: 2018-01-09
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Zhang M;Leitch HG;Tang WWC;Festuccia N;Hall-Ponsele E;Nichols J;Surani MA;Smith A;Chambers I
• 通讯作者: Chambers I

Differential repression of Otx2 underlies the capacity of NANOG and ESRRB to induce germline entry.

• DOI: 10.1016/j.stemcr.2021.11.013
• 发表时间: 2022-01-11
• 期刊: Stem cell reports
• 影响因子: 5.9
• 作者: Vojtek M;Zhang J;Sun J;Zhang M;Chambers I
• 通讯作者: Chambers I