Dynamic transcription factor function in control of pluripotent cell sub-states

控制多能细胞亚状态的动态转录因子功能

• 批准号: MR/L018497/1
• 负责人: Ian Chambers
• 金额: $ 227.57万
• 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=MR%2FL018497%2F1
• 关键词: Dynamic; transcription; factor; function; control

Stem cells have two defining features; they can divide symmetrically to produce cells functionally identical to themselves and can specialise into the more mature cell types that carry out our bodies' functions, a process called differentiation. To preserve a functional stem cell population, self-renewal and differentiation must be balanced.The most versatile mammalian stem cell that can grow in the lab has the ability to differentiate into all adult body cells and is called a pluripotent stem cell. Recently, scientists have found that different types of mouse pluripotent stem cells can be grown in the lab; embryonic stem (ES) cells and epiblast stem cells (EpiSCs). In addition, work from our group has identified molecular heterogeneity within ES cell populations that is related to functional differences between the cells. Specifically, undifferentiated ES cells fluctuate between states in which they have high or low concentrations of a particular transcription factor, which we have named Nanog, and that have, respectively, a greater or lesser likelihood of self-renewal. This fluctuating alteration in the propensity of cells to differentiate may be crucial to balancing the opposing stem cell properties in a population. Therefore, understanding how these reversible states are controlled molecularly is likely to impinge on strategies for achieving predictable, uniform control of differentiation and is thus strategically important.In this research we will examine the function of pluripotency gene regulators at target genes that critically modulate self-renewal efficiency to determine how the pluripotent population is segregated into cells that self-renew efficiently and cells that have a higher likelihood of differentiation. We have identified a small set of 64 genes that alter transcription in response to Nanog activity and that represent good candidate mediators of Nanog function. We will ask how these 64 genes contribute to functional heterogeneity and how gene regulators control the corresponding genes.Aim 1: Test whether Nanog-sensitive genes can fully complement Nanog function. A prominent Nanog-sensitive target, Esrrb can complement several Nanog functions when added back to cells from which Nanog has been removed but cannot fully complement Nanog function. Therefore, we will test the ability of additional candidates to fully compensate for loss of Nanog in combinatorial assays.Several candidate genes are repressed by Nanog, so we will test the ability of reduction in the level of these candidates to compensate for loss of Nanog. Aim 2: Determine the biochemical function of pluripotency gene regulators.We will determine how Nanog fluctuations arise. We will localise gene regulators across the Nanog gene to determine which of these control Nanog and therefore potentially control partitioning between functional subtypes. We have found that Nanog protein represses the Nanog gene and we will ask how this happens to find out if simple rules govern how Nanog switches different genes on and off. Aim 3: Compare pluripotency gene regulator function in vivo and in pluripotent human cells.We will determine whether functional compensations occuring in culture also occur in the mouse embryo. Interestingly, some candidate regulators can reprogramme EpiSCs to an ES cell state. Human ES cells are more like mouse EpiSCs than mouse ES cells, so we will test the ability of our candidates to influence the growth properties of human ES cells. This could beneficially simplify and reduce the cost of human ES cell culture. This work will deliver a deeper, more refined understanding of the mechanisms of action of pluripotency gene regulators in cells in culture and in the embryo.

Stem cells have two defining features; they can divide symmetrically to produce cells functionally identical to themselves and can specialise into the more mature cell types that carry out our bodies' functions, a process called differentiation. To preserve a functional stem cell population, self-renewal and differentiation must be balanced.The most versatile mammalian stem cell that can grow in the lab has the ability to differentiate into all adult body cells and is called a pluripotent stem cell. Recently, scientists have found that different types of mouse pluripotent stem cells can be grown in the lab; embryonic stem (ES) cells and epiblast stem cells (EpiSCs). In addition, work from our group has identified molecular heterogeneity within ES cell populations that is related to functional differences between the cells. Specifically, undifferentiated ES cells fluctuate between states in which they have high or low concentrations of a particular transcription factor, which we have named Nanog, and that have, respectively, a greater or lesser likelihood of self-renewal. This fluctuating alteration in the propensity of cells to differentiate may be crucial to balancing the opposing stem cell properties in a population. Therefore, understanding how these reversible states are controlled molecularly is likely to impinge on strategies for achieving predictable, uniform control of differentiation and is thus strategically important.In this research we will examine the function of pluripotency gene regulators at target genes that critically modulate self-renewal efficiency to determine how the pluripotent population is segregated into cells that self-renew efficiently and cells that have a higher likelihood of differentiation. We have identified a small set of 64 genes that alter transcription in response to Nanog activity and that represent good candidate mediators of Nanog function. We will ask how these 64 genes contribute to functional heterogeneity and how gene regulators control the corresponding genes.Aim 1: Test whether Nanog-sensitive genes can fully complement Nanog function. A prominent Nanog-sensitive target, Esrrb can complement several Nanog functions when added back to cells from which Nanog has been removed but cannot fully complement Nanog function. Therefore, we will test the ability of additional candidates to fully compensate for loss of Nanog in combinatorial assays.Several candidate genes are repressed by Nanog, so we will test the ability of reduction in the level of these candidates to compensate for loss of Nanog. Aim 2: Determine the biochemical function of pluripotency gene regulators.We will determine how Nanog fluctuations arise. We will localise gene regulators across the Nanog gene to determine which of these control Nanog and therefore potentially control partitioning between functional subtypes. We have found that Nanog protein represses the Nanog gene and we will ask how this happens to find out if simple rules govern how Nanog switches different genes on and off. Aim 3: Compare pluripotency gene regulator function in vivo and in pluripotent human cells.We will determine whether functional compensations occuring in culture also occur in the mouse embryo. Interestingly, some candidate regulators can reprogramme EpiSCs to an ES cell state. Human ES cells are more like mouse EpiSCs than mouse ES cells, so we will test the ability of our candidates to influence the growth properties of human ES cells. This could beneficially simplify and reduce the cost of human ES cell culture. This work will deliver a deeper, more refined understanding of the mechanisms of action of pluripotency gene regulators in cells in culture and in the embryo.

项目成果

Dynamic changes in Sox2 spatio-temporal expression promote the second cell fate decision through Fgf4/Fgfr2 signaling in preimplantation mouse embryos.

• DOI: 10.1042/bcj20170418
• 发表时间: 2018-03-20
• 期刊: The Biochemical journal
• 作者: Mistri TK;Arindrarto W;Ng WP;Wang C;Lim LH;Sun L;Chambers I;Wohland T;Robson P
• 通讯作者: Robson P

Distinct Signaling Requirements for the Establishment of ESC Pluripotency in Late-Stage EpiSCs.

• DOI: 10.1016/j.celrep.2016.03.073
• 发表时间: 2016-04-26
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Illich DJ;Zhang M;Ursu A;Osorno R;Kim KP;Yoon J;Araúzo-Bravo MJ;Wu G;Esch D;Sabour D;Colby D;Grassme KS;Chen J;Greber B;Höing S;Herzog W;Ziegler S;Chambers I;Gao S;Waldmann H;Schöler HR
• 通讯作者: Schöler HR

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

Distinct SoxB1 networks are required for naïve and primed pluripotency.

• DOI: 10.7554/elife.27746
• 发表时间: 2017-12-19
• 期刊: eLife
• 影响因子: 7.7
• 作者: Corsinotti A;Wong FC;Tatar T;Szczerbinska I;Halbritter F;Colby D;Gogolok S;Pantier R;Liggat K;Mirfazeli ES;Hall-Ponsele E;Mullin NP;Wilson V;Chambers I
• 通讯作者: Chambers I

Functional Dissection of the Enhancer Repertoire in Human Embryonic Stem Cells.

• DOI: 10.1016/j.stem.2018.06.014
• 发表时间: 2018-08-02
• 期刊: Cell stem cell
• 影响因子: 23.9
• 作者: Barakat TS;Halbritter F;Zhang M;Rendeiro AF;Perenthaler E;Bock C;Chambers I
• 通讯作者: Chambers I