Canonical Wnt Pathway Controls mESC Self-Renewal Through Inhibition of Spontaneous Differentiation via β-Catenin/TCF/LEF Functions

Aulicino, Francesco; Pedone, Elisa; Sottile, Francesco; Lluı́s, Frederic; Marucci, Lucia; Cosma, María Pía

doi:10.1016/j.stemcr.2020.07.019

Cited by 27 publications

(36 citation statements)

References 31 publications

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“…teratomas from the mutants with the highest β -catenin transcriptional activity showed major differentiation defects in the neuroectoderm, dorsal mesoderm and endoderm lineages. Of note, results in (Kielman et al, 2002) (Lyashenko et al, 2011, Wray et al, 2011, Faunes et al, 2013, Aulicino et al, 2020, while suggesting that specific β -catenin perturbations cause a bias towards the endoderm lineage, in line with Lef-1 related results previously reported (Ye et al, 2017).…”

Section: Introductionsupporting

confidence: 73%

“…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%

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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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Section: Introductionsupporting

confidence: 73%

Section: Introductionmentioning

confidence: 99%

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

Pedone

Failli

Gambardella

et al. 2020

Preprint

Self Cite

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“…is initiated upon binding of Wnt ligands with the membrane receptor complex known as Frizzled/low-density lipoprotein receptor-related protein 5/6 (FZD/LRP) [59]. Interactions between Wnt ligands and FZD/LRP5/6 trigger disheveled (DSH) that switches off glycogen synthase kinase 3 β (GSK-3β) and enables β-catenin protein to be translocated into the nucleus, whereby it complexes with the binding T cell factor/lymphoid enhancer-binding factor (TCF/LEF) to stimulate the expression of target genes involved in osteogenesis, including RUNX2 [60,61] (Figure 3). Several Wnt ligands, including Wnt4, Wnt5a, Wnt6, Wnt10a and Wnt10b, are involved in initiating the Wnt/β-catenin cascade and stimulation of osteoblast differentiation [62][63][64].…”

Section: Msc Signaling Pathways In Osteogenic Differentiationmentioning

confidence: 99%

MicroRNAs Modulate Signaling Pathways in Osteogenic Differentiation of Mesenchymal Stem Cells

Mazziotta

Lanzillotti

Iaquinta

et al. 2021

IJMS

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Mesenchymal stem cells (MSCs) have been identified in many adult tissues and they have been closely studied in recent years, especially in view of their potential use for treating diseases and damaged tissues and organs. MSCs are capable of self-replication and differentiation into osteoblasts and are considered an important source of cells in tissue engineering for bone regeneration. Several epigenetic factors are believed to play a role in the osteogenic differentiation of MSCs, including microRNAs (miRNAs). MiRNAs are small, single-stranded, non-coding RNAs of approximately 22 nucleotides that are able to regulate cell proliferation, differentiation and apoptosis by binding the 3′ untranslated region (3′-UTR) of target mRNAs, which can be subsequently degraded or translationally silenced. MiRNAs control gene expression in osteogenic differentiation by regulating two crucial signaling cascades in osteogenesis: the transforming growth factor-beta (TGF-β)/bone morphogenic protein (BMP) and the Wingless/Int-1(Wnt)/β-catenin signaling pathways. This review provides an overview of the miRNAs involved in osteogenic differentiation and how these miRNAs could regulate the expression of target genes.

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“…In addition, our data indicated that most general naive markers in L-ESCs were clustered with 2i/L-ESCs ( Figures S2 A and S2B), and primed markers in L-ESCs exhibit intermediate state between 2i/L-ESCs and EpiSCs ( Figure S2 C). Furthermore, β-CATENIN and ERK play central roles in balancing differentiation and self-renewal ( Aulicino et al., 2020 ); hence, we tested the β-CATENIN and ERK levels in L-ESCs. Notably, some WNT signaling-related genes and the protein level of β-CATENIN were differentially expressed in L-ESCs compared with 2i/L-ESCs ( Figures S2 D and S2E).…”

Section: Resultsmentioning

confidence: 99%

DNMTs Play an Important Role in Maintaining the Pluripotency of Leukemia Inhibitory Factor-Dependent Embryonic Stem Cells

et al. 2021

Stem Cell Reports

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Summary Naive pluripotency can be maintained in medium with two inhibitors plus leukemia inhibitory factor (2i/LIF) supplementation, which primarily affects canonical WNT, FGF/ERK, and JAK/STAT3 signaling. However, whether one of these three supplements alone is sufficient to maintain naive self-renewal remains unclear. Here we show that LIF alone in medium is sufficient for adaptation of 2i/L-ESCs to embryonic stem cells (ESCs) in a hypermethylated state (L-ESCs). Global transcriptomic analysis shows that L-ESCs are close to 2i/L-ESCs and in a stable state between naive and primed pluripotency. Notably, our results demonstrate that DNA methyltransferases (DNMTs) play an important role in LIF-dependent mouse ESC adaptation and self-renewal. LIF-dependent ESC adaptation efficiency is significantly increased in serum treatment and reduced in Dnmt3a or Dnmt3l knockout ESCs. Importantly, unlike epiblast stem cells, L-ESCs contribute to somatic tissues and germ cells in chimeras. L-ESCs cultured under such simple conditions as in this study would provide a more conducive platform to clarify the molecular mechanism of ESCs in in vitro culture.

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Canonical Wnt Pathway Controls mESC Self-Renewal Through Inhibition of Spontaneous Differentiation via β-Catenin/TCF/LEF Functions

Cited by 27 publications

References 31 publications

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

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

MicroRNAs Modulate Signaling Pathways in Osteogenic Differentiation of Mesenchymal Stem Cells

DNMTs Play an Important Role in Maintaining the Pluripotency of Leukemia Inhibitory Factor-Dependent Embryonic Stem Cells

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