MRAS GTPase is a novel stemness marker that impacts mouse embryonic stem cell plasticity and <i>Xenopus</i> embryonic cell fate

Mathieu, M.; Faucheux, Claude; Saucourt, Claire; Soulet, Fabienne; Gauthereau, Xavier; Fédou, Sandrine; Trouillas, Marina; Thézé, Nadine; Thiébaud, Pierre; Bœuf, Hélène

doi:10.1242/dev.091082

Cited by 12 publications

(7 citation statements)

References 72 publications

(102 reference statements)

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“…There are two categories of Lifind genes: those expressed after LIF induction specifically in cells depleted of LIF for 24 hours ( speLifind) like Kruppel-like factor 5 ( Klf5 ) and those induced in a more general way, in immature as well as differentiated cells ( pleioLifind ) like JunB . [ 42 – 44 ]. Klf5 , a transcription factor of the Kruppel gene family, well conserved upon evolution, has been shown to be essential for the derivation of mESCs from Inner Cell Mass (ICM) as well as for the maintenance of mESCs cell pluripotency in vitro [ 45 – 48 ].…”

Section: Introductionmentioning

confidence: 99%

Murine Embryonic Stem Cell Plasticity Is Regulated through Klf5 and Maintained by Metalloproteinase MMP1 and Hypoxia

et al. 2016

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Mouse embryonic stem cells (mESCs) are expanded and maintained pluripotent in vitro in the presence of leukemia inhibitory factor (LIF), an IL6 cytokine family member which displays pleiotropic functions, depending on both cell maturity and cell type. LIF withdrawal leads to heterogeneous differentiation of mESCs with a proportion of the differentiated cells apoptosising. During LIF withdrawal, cells sequentially enter a reversible and irreversible phase of differentiation during which LIF addition induces different effects. However the regulators and effectors of LIF–mediated reprogramming are poorly understood. By employing a LIF-dependent ‘plasticity’ test, that we set up, we show that Klf5, but not JunB is a key LIF effector. Furthermore PI3K signaling, required for the maintenance of mESC pluripotency, has no effect on mESC plasticity while displaying a major role in committed cells by stimulating expression of the mesodermal marker Brachyury at the expense of endoderm and neuroectoderm lineage markers. We also show that the MMP1 metalloproteinase, which can replace LIF for maintenance of pluripotency, mimics LIF in the plasticity window, but less efficiently. Finally, we demonstrate that mESCs maintain plasticity and pluripotency potentials in vitro under hypoxic/physioxic growth conditions at 3% O2 despite lower levels of Pluri and Master gene expression in comparison to 20% O2.

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Section: Introductionmentioning

confidence: 99%

Murine Embryonic Stem Cell Plasticity Is Regulated through Klf5 and Maintained by Metalloproteinase MMP1 and Hypoxia

et al. 2016

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“…Relative quantification was done by calculating the 2ΔΔCt value. Data were normalized using the Hprt mRNA, known to remain constant in the experimental conditions .…”

Section: Methodsmentioning

confidence: 99%

Bioenergetic Changes Underline Plasticity of Murine Embryonic Stem Cells

et al. 2019

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Murine embryonic stem cells (mESCs) are endowed by a time-dependent window of plasticity during their early commitment steps. Indeed, while mESCs deprived of leukemia inhibitory factor (LIF) for 24 hours revert to their naive pluripotent state after subsequent LIF readdition, cells deprived of LIF for 48 hours are no longer efficient in reverting, upon LIF addition, and undergo irreversible differentiation. We investigated undisclosed bioenergetic profiles of early mESCderived committed cells versus their undifferentiated states in order to reveal specific bioenergetic changes associated with mESC plasticity. Multiparametric bioenergetic analysis revealed that pluripotent (+LIF) and reversibly committed cells (−LIF24h) are energetically flexible, depending on both oxidative phosphorylation (OXPHOS) and glycolysis. They exhibit high mitochondrial respiration in the presence of the main energetic substrates and can also rely on glycolysis in the presence of OXPHOS inhibitor. Inhibition of the glycolysis or mitochondrial respiration does not change drastically the expression of pluripotency genes, which remain well expressed. In addition, cells treated with these inhibitors keep their capacity to differentiate efficiently upon embryoid bodies formation. Transition from metabolically active mESCs to irreversibly committed cells is associated with a clear change in mitochondrial network morphology, to an increase of adenosine triphosphate (ATP) produced from glycolysis and a decline of ATP turnover and of the mitochondrial activity without change in the mitochondrial mass. Our study pointed that plasticity window of mESCs is associated with the bivalent energetic metabolism and potency to shift to glycolysis or OXPHOS on demand. LIF removal provokes glycolytic metabolic orientation and consecutive loss of the LIFdependent reversion of cells to the pluripotent state. STEM CELLS 2019;37:463-475 SIGNIFICANCE STATEMENTMurine embryonic stem cells rely on leukemia inhibitory factor (LIF) cytokine to maintain their naïve pluripotent state. They undergo differentiation by simple LIF removal with a 48-hour period of plasticity window in which cells can maintain a pluripotent behavior upon LIF readdition. The study has established key point of metabolic changes, by multiparametric bioenergetic analysis, during this period of transition from naïve pluripotent toward committed cell states. Gene expression profiles and functional tests allow to conclude that cells maintained with LIF or depleted of LIF for 24 hours are energetically flexible, depending on both oxidative phosphorylation and glycolysis, whereas after 48 hours of LIF withdrawal, they are mainly glycolytic.

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“…Of all the Ras family members, Mras is the only gene whose expression is limited to undifferentiated mESCs. Prior to differentiation, expression is controlled by the cytokine LIF, and as such Mras is described as a marker of stemness (Mathieu et al 2013). Persistent Mras down-regulation by removal of LIF affects the normal balance of expression of core pluripotency markers such as OCT4 and CAECAM1, and overexpression of MRAS leads to sustained expression of OCT4 and NANOG.…”

Section: Other Functions Of Mrasmentioning

confidence: 99%

“…Persistent Mras down-regulation by removal of LIF affects the normal balance of expression of core pluripotency markers such as OCT4 and CAECAM1, and overexpression of MRAS leads to sustained expression of OCT4 and NANOG. In Xenopus, Mras is expressed throughout the embryo, maintaining pluripotency, until the blastula stage, and as in mice, it is required for neuronal differentiation, which indicates that this protein has conserved functions across vertebrates (Mathieu et al 2013).…”

Section: Other Functions Of Mrasmentioning

confidence: 99%

MRAS: A Close but Understudied Member of the RAS Family

Young

Rodríguez‐Viciana

2018

Cold Spring Harb Perspect Med

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MRAS is the closest relative to the classical RAS oncoproteins and shares most regulatory and effector interactions. However, it also has unique functions, including its ability to function as a phosphatase regulatory subunit when in complex with SHOC2 and protein phosphatase 1 (PP1). This phosphatase complex regulates a crucial step in the activation cycle of RAF kinases and provides a key coordinate input required for efficient ERK pathway activation and transformation by RAS. MRAS mutations rarely occur in cancer but deregulated expression may play a role in tumorigenesis in some settings. Activating mutations in MRAS (as well as SHOC2 and PP1) do occur in the RASopathy Noonan syndrome, underscoring a key role for MRAS within the RAS-ERK pathway. MRAS also has unique roles in cell migration and differentiation and has properties consistent with a key role in the regulation of cell polarity. Further investigations should shed light on what remains a relatively understudied RAS family member.

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MRAS GTPase is a novel stemness marker that impacts mouse embryonic stem cell plasticity and Xenopus embryonic cell fate

Cited by 12 publications

References 72 publications

Murine Embryonic Stem Cell Plasticity Is Regulated through Klf5 and Maintained by Metalloproteinase MMP1 and Hypoxia

Murine Embryonic Stem Cell Plasticity Is Regulated through Klf5 and Maintained by Metalloproteinase MMP1 and Hypoxia

Bioenergetic Changes Underline Plasticity of Murine Embryonic Stem Cells

MRAS: A Close but Understudied Member of the RAS Family

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