Transcription factor control of dynamic transitions within and beyond pluripotency

多能性内外动态转变的转录因子控制

• 批准号: MR/T003162/1
• 负责人: Ian Chambers
• 金额: $ 260.77万
• 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: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FT003162%2F1
• 关键词: Transcription; factor; control; dynamic; transitions

Cell identity depends on the action of transcription factors and environmental signals that together read the genome. The mechanisms by which cell identity changes during development are of particular interest to fundamental developmental and stem cell biology. In particular, pluripotent cells, which can both self-renew and differentiate to give rise to all lineages in vitro and in vivo, are ideally suited to studying maintenance and changes in cell identity. Application of such knowledge is crucial to the design of robust protocols for in vitro differentiation of cells free from germline tumour-initiating cells for use in cell transplantation and drug discovery. Pluripotent stem cell self-renewal is governed by a pluripotency gene regulatory network centred on the transcription factors OCT4, SOX2 and NANOG. While considerable advances have been made in identifying additional pluripotency gene regulatory network components and in analysing global chromatin binding by transcription factors, these powerful approaches do not tell us how cell decisions are actually made. To better understand the distinction between self-renewal and differentiation we will use precise mechanistic analysis to determine how individual transcription factors affect the operation of the pluripotency gene regulatory network.Pluripotent cells pass through successive states during differentiation. Segregation of germline and somatic lineages occurs from a transitional 'formative' pluripotent state. A key unanswered question is why many pluripotency transcription factors (including OCT4, SOX2 and NANOG) also operate in the germline gene regulatory network and are critical for primordial germ cell (PGC) function. As these cell types have arguably the most radically divergent potencies, this is paradoxical. By studying OCT4, SOX2 and NANOG and by doing so in cells at distinct stages during the transition from naïve pluripotent embryonic stem cells (ESCs) to formative epiblast-like cells (EpiLCs) and from EpiLCs to PGCs, we aim to resolve this paradox and to reveal principles at the very foundation of phenotypic cell identity. Our studies will extend knowledge from the mouse to address germline entry in humans. This proposal has three aims, focused on distinct aspects of pluripotent cell function:1. How is pluripotency specified and lost in vivo?2. How do transcription factors act in the maintenance and loss of naïve pluripotency in vitro?3. What regulatory interactions determine the efficiency of entry to the germline?Delivering the above aims will advance the field by assessing how transcription factors operate as an ensemble to control ESC identity, and by revealing how transcription factors are repurposed at distinct developmental stages. This will break new ground by identifying key mechanisms by which cells exit specific pluripotent states, particularly as they enter the germline, or initiate entry into the primitive streak. Together, this knowledge will provide the insights needed to rigorously command the uniform differentiation of pluripotent cells demanded by future recipients of regenerative medicine strategies.

细胞的身份取决于转录因子和环境信号的作用，它们共同阅读基因组。细胞身份在发育过程中发生变化的机制对基础发育和干细胞生物学特别感兴趣。特别是，多能细胞，它可以自我更新和分化，在体外和体内产生所有的谱系，是理想的适合研究细胞特性的维持和变化。这些知识的应用对于设计稳健的方案至关重要，该方案用于体外分化不含种系肿瘤起始细胞的细胞，以用于细胞移植和药物发现。多能干细胞的自我更新由以转录因子OCT4、SOX 2和NANOG为中心的多能性基因调控网络控制。虽然在识别额外的多能性基因调控网络组件和分析转录因子的全局染色质结合方面取得了相当大的进展，但这些强大的方法并不能告诉我们细胞的决定实际上是如何做出的。为了更好地理解自我更新和分化之间的区别，我们将使用精确的机制分析来确定单个转录因子如何影响多能性基因调控网络的运作。生殖系和体细胞谱系的分离发生在过渡的“形成性”多能状态。一个关键的未回答的问题是为什么许多多能性转录因子（包括OCT 4，SOX 2和NANOG）也在生殖系基因调控网络中运作，并且对原始生殖细胞（PGC）功能至关重要。由于这些细胞类型可以说是最根本不同的潜力，这是矛盾的。通过研究OCT 4，SOX 2和NANOG，并在从幼稚多能胚胎干细胞（ESC）到形成性外胚层样细胞（EpiLC）以及从EpiLC到PGC的过渡过程中的不同阶段的细胞中进行研究，我们的目标是解决这个悖论，并揭示表型细胞身份的基础原则。我们的研究将扩展小鼠的知识，以解决人类的生殖系进入。该提案有三个目标，侧重于多能细胞功能的不同方面：1。多能性是如何在体内指定和丢失的？2.转录因子如何在体外维持和丧失幼稚多能性中发挥作用？3.哪些监管相互作用决定了进入种系的效率？实现上述目标将通过评估转录因子如何作为一个整体来控制ESC身份，以及通过揭示转录因子如何在不同的发育阶段被重新利用来推进该领域。这将通过确定细胞退出特定多能状态的关键机制来开辟新天地，特别是当它们进入生殖系或开始进入原始条纹时。总之，这些知识将提供所需的见解，以严格指挥再生医学战略的未来接受者所要求的多能细胞的均匀分化。

项目成果

Phosphorylation of NANOG by casein kinase I regulates embryonic stem cell self-renewal.

• DOI: 10.1002/1873-3468.13969
• 发表时间: 2021-01
• 期刊: FEBS letters
• 影响因子: 3.5
• 作者: Mullin NP;Varghese J;Colby D;Richardson JM;Findlay GM;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

Loss of Resf1 reduces the efficiency of embryonic stem cell self-renewal and germline entry.

RESF1的丢失降低了胚胎干细胞自我更新和种系的效率。

• DOI: 10.26508/lsa.202101190
• 发表时间: 2021-12
• 期刊: Life science alliance
• 影响因子: 4.4
• 作者: Vojtek M;Chambers I
• 通讯作者: Chambers I