β-Catenin and Associated Proteins Regulate Lineage Differentiation in Ground State Mouse Embryonic Stem Cells

Fang, Tao; Soffers, Jelly H.M.; Hu, Deqing; Chen, Shiyuan; Gào, Xin; Zhang, Ying; Zhao, Chongbei; Smith, Sarah E.; Unruh, Jay R.; Zhang, Da; Tsuchiya, Dai; Venkatraman, Aparna; Zhao, Meng; Li, Zhenrui; Qian, Pengxu; Parmely, Tari; Washburn, Michael P.; Florens, Laurence; Perry, John M.; Zeitlinger, Julia; Workman, Jerry L.; Li, Linheng

doi:10.1016/j.stemcr.2020.07.018

Cited by 11 publications

(18 citation statements)

References 48 publications

(62 reference statements)

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“…Secondly, β-CATENIN can act as both an activator and repressor of downstream targets. A similar conclusion was also drawn based on the study from Tao et al (20) and this may be dependent on the influence and presence of additional interactors.…”

Section: Discussionsupporting

confidence: 78%

“…The significant reduction in the activity of β-CATENIN/TCF in Cdh1 -/- mESCs prompted us to check the expression of β-CATENIN targets in these cells. We overlaid the ChIP-seq data published by Tao et al (20) with our RNA-seq data to analyse the expression profile of bona fide β-CATENIN targets. 23% of differentially expressed genes (both up- and down-regulated) were β-CATENIN targets (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In a parallel study, it was reported that β-catenin has repressive functions upon engagement with factors such as E2F6, HMGA2, and HP1ϒ, preventing the differentiation of mESCs towards a neural lineage. These findings reveal a new role for β-CATENIN whereby it not only activates the transcription of core pluripotency factors, but can also repress expression of genes associated with lineage differentiation to maintain the ground state of mESCs (20).…”

Section: Introductionmentioning

confidence: 95%

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E-cadherin regulates the stability and transcriptional activity of β-catenin in embryonic stem cells

Bhattacharyya

Mote

Freimer

et al. 2021

Preprint

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E-CADHERIN is abundantly expressed in embryonic stem cells (ESCs) and plays an important role in the maintenance of cell-cell adhesions. However, the exact function of this molecule beyond cell adhesion, in the context of cell fate decisions is largely unknown. Using mouse ESCs (mESCs), we demonstrate that E-CADHERIN and β-CATENIN interact at the membrane and continue to do so upon internalization within the cell. Knockout of the gene encoding E-CADHERIN, Cdh1, in mESCs resulted in a failure to form tight colonies, accompanied by altered expression of differentiation markers, and retention of pluripotency factor expression during differentiation. Interestingly, Cdh1-/- mESCs showed a dramatic reduction in β-CATENIN levels. Transcriptional profiling of Cdh1-/- mESCs displayed a significant alteration in the expression of a subset of β-CATENIN targets, in a cell-state dependent manner. While treatment with a pharmacological inhibitor against GSK3β could rescue levels of β-CATENIN in Cdh1-/- mESCs, expression of downstream targets were altered in a context-dependent manner, indicating an additional layer of regulation within this subset. Together, our results reveal the existence of a cell-state-dependent regulation of β-CATENIN and its transcriptional targets in an E-CADHERIN dependent manner. Our findings hint at hitherto unknown roles played by E-CADHERIN in regulating the activity of β-CATENIN in ESCs.

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Section: Discussionsupporting

confidence: 78%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

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E-cadherin regulates the stability and transcriptional activity of β-catenin in embryonic stem cells

Bhattacharyya

Mote

Freimer

et al. 2021

Preprint

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“…Bisulfite sequencing confirmed that the promoters of E2F6 target genes are hypomethylated in 2i ESCs (Supplementary Fig. 5b ) and E2F6 binding to germline genes is unchanged in 2i conditions as shown by a recent E2F6 ChIP-seq in 2i ESCs 41 (Supplementary Fig. 5c, d ).…”

Section: Resultsmentioning

confidence: 52%

E2F6 initiates stable epigenetic silencing of germline genes during embryonic development

et al. 2021

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In mouse development, long-term silencing by CpG island DNA methylation is specifically targeted to germline genes; however, the molecular mechanisms of this specificity remain unclear. Here, we demonstrate that the transcription factor E2F6, a member of the polycomb repressive complex 1.6 (PRC1.6), is critical to target and initiate epigenetic silencing at germline genes in early embryogenesis. Genome-wide, E2F6 binds preferentially to CpG islands in embryonic cells. E2F6 cooperates with MGA to silence a subgroup of germline genes in mouse embryonic stem cells and in embryos, a function that critically depends on the E2F6 marked box domain. Inactivation of E2f6 leads to a failure to deposit CpG island DNA methylation at these genes during implantation. Furthermore, E2F6 is required to initiate epigenetic silencing in early embryonic cells but becomes dispensable for the maintenance in differentiated cells. Our findings elucidate the mechanisms of epigenetic targeting of germline genes and provide a paradigm for how transient repression signals by DNA-binding factors in early embryonic cells are translated into long-term epigenetic silencing during mouse development.

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“…These negative feedback loops contribute to the emergence of nonlinear dynamics in the Wnt/β-catenin pathway, proved to be important in different biological and developmental aspects (see for a review), ESCs pluripotency and somatic cell reprogramming (Marucci et al, 2014, Aulicino et al, 2014, Ho et al, 2013, Kimura et al, 2016, Lluis et al, 2008. The role of the canonical Wnt pathway in early in vivo developmental stages and the requirement of its activation for ESC self-renewal have been a matter of intense research (Anton et al, 2007, Ying et al, 2008, Soncin et al, 2009, Wagner et al, 2010, Lyashenko et al, 2011, Wray et al, 2011, Faunes et al, 2013, Chatterjee et al, 2015, Ortmann et al, 2020, Tao et al, 2020, Theka et al, 2019, Aulicino et al, 2020. Pluripotency incompetence of β -catenin -/-ECSs has been reported in two independent studies (Anton et al, 2007, Wagner et al, 2010; this phenotype was contradicted later using newly generated β -catenin -/cell lines, which showed selfrenewal in both serum and 2i+LIF (hereafter called 2i/L), but presented some differentiation defects when LIF-deprived (Wray et al, 2011, Lyashenko et al, 2011, Aulicino et al, 2020.…”

Section: Introductionmentioning

confidence: 99%

β-catenin perturbations control differentiation programs in mouse embryonic stem cells

Pedone

Failli

Gambardella

et al. 2020

Preprint

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12The Wnt/β-catenin pathway is involved in development, cancer and embryonic stem 13 cell (ESC) maintenance; its dual role in stem cell self-renewal and differentiation is still 14 controversial. Here, we applied an elegant in vitro system enabling conditional β-15 catenin control in β-catenin null mouse ESCs. We report that moderate induction of 16 exogenous β-catenin enhances epiblast stem cell (EpiSC) derivation in vitro. Using a 17 different genetic model and β-catenin chemical modulation, we derived a new protocol 18 for ESCs to EpiSCs differentiation, based on NDiff227 and the GSK3α/β inhibitor 19Chiron, that is more efficient than standard ActivinA/Fgf2-based protocols. Finally, we 20 report that moderate β-catenin levels favour early stem cell commitment towards 21 mesoderm if the protein is overexpressed only in the 'ground state' of pluripotency 22 conditions, or endoderm if the overexpression is maintained during the differentiation, 23unravelling the controversial role of this signalling pathway at the exit from 24 pluripotency. 25 26 Introduction 27Pluripotent Cells (PCs) are characterized by indefinite proliferative and differentiation 28 potential and their identity is determined by the balance between signals promoting 29 self-renewal and differentiation. The first step for stem cell differentiation is the exit 30 from the pluripotent state, tightly controlled by underlying gene regulatory network 31 dynamics which can drive specific lineage commitment. During murine development 32 in vivo, embryonic stem cells (ESCs), that represent the naïve pluripotent state of the 33 early epiblast 1-3 , convert into the late epiblast and finally in terminally differentiated somatic cells. ESCs can be derived from the pre-implantation epiblast and used to 35 study cell pluripotency and differentiation, providing an excellent in vitro system for 36 understanding signalling pathway interplay in cell fate decision making. 37In serum-based cultures, mouse ESCs (hereafter called ESCs) are heterogeneous for 38 the expression of pluripotency genes 1,2,4-10 , while, when cultured in serum-free media 39 supplemented with inhibitors of MEK1/2 (PD) and GSK3α/β (Chiron) and in presence 40 or not of the Leukaemia Inhibitory Factor-LIF (2i or 2i+LIF) 3 , a uniform self-renewal 41 condition known as 'ground state' of pluripotency is established; it is characterized by 42 homogenous gene expression 11-14 , genome demethylation 15-17 and stable naïve 43 pluripotency 18,19 . Mouse epiblast stem cells (hereafter called EpiSCs) also represent 44 a relevant in vitro model because of their similarities with embryonic stem cells of 45 human origin 20 . EpiSCs, derived from the post-implantation epiblast, are also capable 46 of differentiating in all the germ-layers 20,21 ; however, they differ from ESCs in 47 morphology, clonogenicity, gene expression, epigenome status and, most importantly, 48 ability to contribute to chimaeras 14,[20][21][22] . EpiSCs require ActivinA and the fibroblast 49 growth factor 2 (FGF2) 20,21 for in vitr...

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β-Catenin and Associated Proteins Regulate Lineage Differentiation in Ground State Mouse Embryonic Stem Cells

Cited by 11 publications

References 48 publications

E-cadherin regulates the stability and transcriptional activity of β-catenin in embryonic stem cells

E-cadherin regulates the stability and transcriptional activity of β-catenin in embryonic stem cells

E2F6 initiates stable epigenetic silencing of germline genes during embryonic development

β-catenin perturbations control differentiation programs in mouse embryonic stem cells

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