Inhibition of GSK3β enhances both adhesive and signalling activities of β‐catenin in mouse embryonic stem cells

Sineva, G S; Поспелов, В. А.

doi:10.1042/bc20100016

Cited by 59 publications

(66 citation statements)

References 35 publications

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“…Our results suggest that the activation of PI3K in the later-stage embryo is a part of a signaling pathway involved in insulin's stimulation of epiblast cell numbers. An intermediate of the PI3K signaling pathway, GSK3, has been shown to decrease Nanog transcription and retention of pluripotency via inactivation of b-catenin [66][67][68], Hedgehog [69,70], and c-Myc [71,72]. Additionally, active GSK3 protects the intracellular domain of Notch from degradation [73], increasing differentiation [74].…”

Section: Discussionmentioning

confidence: 99%

Insulin Increases Epiblast Cell Number of In Vitro Cultured Mouse Embryos via the PI3K/GSK3/p53 Pathway

Campbell

Nottle

Vassiliev

et al. 2012

Stem Cells and Development

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Authors may also deposit this version on his/her funder's or funder's designated repository at the funder's request or as a result of a legal obligation, provided it is not made publicly available until 12 months after official publication. th March 2013Insulin High-quality embryos give rise to embryonic stem cells (ESCs) at greater efficiencies than poor-quality embryos. However, most embryos available for human ESC derivation are of a reduced quality as a result of culture in relatively simple media up to 10 years earlier, before cryopreservation, or before compaction. In the present study, we used a mouse model to determine whether a culture with insulin from the 8-cell stage could increase the number of ESC progenitor epiblast cells in blastocysts, as well as endeavor to determine the molecular mechanism of the insulin's effect. Culture in media containing 1.7 rM insulin increased epiblast cell number (determined by Oct4 and Nanog co-expression), and proportion in day 6 blastocysts. The inhibition of phosphoinositide 3 kinase (PI3K) (via LY294002), an early second messenger of the insulin receptor, blocked this effect. The inhibition of glycogen synthase kinase 3 (GSK3) or p53, 2 s messengers inactivated by insulin signaling (via CT99021 or pifithrin-a, respectively), increased epiblast cell numbers. When active, GSK3 and p53 block the transcription of Nanog, which is important for maintaining pluripotency. A simultaneous inhibition of GSK3 and p53 had no synergistic effects on epiblast cell number. The induced activation of GSK3 and p53, via the inhibition of proteins responsible for their inactivation (PKA via H-89 and SIRT-1 via nicotinamide, respectively), blocked the insulin's effect on the epiblast.From our findings, we conclude that insulin increases epiblast cell number via the activation of PI3K, which ultimately inactivates GSK3 and p53. Furthermore, we suggest that the inclusion of insulin in culture media could be used as a strategy for increasing the efficiency with which the ESC lines can be derived from cultured embryos.

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Section: Discussionmentioning

confidence: 99%

Insulin Increases Epiblast Cell Number of In Vitro Cultured Mouse Embryos via the PI3K/GSK3/p53 Pathway

Campbell

Nottle

Vassiliev

et al. 2012

Stem Cells and Development

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“…To demonstrate that the robust expansion obtained with BIO treatment was due to Gsk3 inhibition and not due to off-target effects, we repeated a similar series of experiments using two other known analogous Gsk3 inhibitors: 1-azakenpaullone (AZA) (Kunick et al, 2004;Qyang et al, 2007) and CHIR-99021 (CHI) (Sineva and Pospelov, 2010;Ying et al, 2008). AZA and CHI were shown to selectively inhibit the β-isoform of cytoplasmic Gsk3.…”

Section: Wnt Signaling Promotes the Expansion Of Escderived Fhf-and Smentioning

confidence: 99%

Wnt/β-catenin signaling directs the regional expansion of first and second heart field-derived ventricular cardiomyocytes

et al. 2013

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SUMMARYIn mammals, cardiac development proceeds from the formation of the linear heart tube, through complex looping and septation, all the while increasing in mass to provide the oxygen delivery demands of embryonic growth. The developing heart must orchestrate regional differences in cardiomyocyte proliferation to control cardiac morphogenesis. During ventricular wall formation, the compact myocardium proliferates more vigorously than the trabecular myocardium, but the mechanisms controlling such regional differences among cardiomyocyte populations are not understood. Control of definitive cardiomyocyte proliferation is of great importance for application to regenerative cell-based therapies. We have used murine and human pluripotent stem cell systems to demonstrate that, during in vitro cellular differentiation, early ventricular cardiac myocytes display a robust proliferative response to β-cateninmediated signaling and conversely accelerate differentiation in response to inhibition of this pathway. Using gain-and loss-of-function murine genetic models, we show that β-catenin controls ventricular myocyte proliferation during development and the perinatal period. We further demonstrate that the differential activation of the Wnt/β-catenin signaling pathway accounts for the observed differences in the proliferation rates of the compact versus the trabecular myocardium during normal cardiac development. Collectively, these results provide a mechanistic explanation for the differences in localized proliferation rates of cardiac myocytes and point to a practical method for the generation of the large numbers of stem cell-derived cardiac myocytes necessary for clinical applications.

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“…3A), confirming the activity of BIX01294 in promoting specific chromatin modifications. For the other molecules used in this study, we performed positive control experiments of previously described activities [49,[53][54][55] to ensure that they were functionally active despite their inability to promote expression of cardiac progenitor genes. For example, both CHIR99021 and Wnt3a provoked the nuclear (Fig.…”

Section: Upregulation Of Mesodermal and Cardiac Progenitor Markers Inmentioning

confidence: 99%

The Histone Methyltransferase Inhibitor BIX01294 Enhances the Cardiac Potential of Bone Marrow Cells

Mezentseva

Yang²,

Kaur³

et al. 2013

Stem Cells and Development

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Bone marrow (BM) has long been considered a potential stem cell source for cardiac repair due to its abundance and accessibility. Although previous investigations have generated cardiomyocytes from BM, yields have been low, and far less than produced from ES or induced pluripotent stem cells (iPSCs). Since differentiation of pluripotent cells is difficult to control, we investigated whether BM cardiac competency could be enhanced without making cells pluripotent. From screens of various molecules that have been shown to assist iPSC production or maintain the ES cell phenotype, we identified the G9a histone methyltransferase inhibitor BIX01294 as a potential reprogramming agent for converting BM cells to a cardiac-competent phenotype. BM cells exposed to BIX01294 displayed significantly elevated expression of brachyury, Mesp1, and islet1, which are genes associated with embryonic cardiac progenitors. In contrast, BIX01294 treatment minimally affected ectodermal, endodermal, and pluripotency gene expression by BM cells. Expression of cardiac-associated genes Nkx2.5, GATA4, Hand1, Hand2, Tbx5, myocardin, and titin was enhanced 114,76, 276,46, 635, 123, and 5-fold in response to the cardiogenic stimulator Wnt11 when BM cells were pretreated with BIX01294. Immunofluorescent analysis demonstrated that BIX01294 exposure allowed for the subsequent display of various muscle proteins within the cells. The effect of BIX01294 on the BM cell phenotype and differentiation potential corresponded to an overall decrease in methylation of histone H3 at lysine9, which is the primary target of G9a histone methyltransferase. In summary, these data suggest that BIX01294 inhibition of chromatin methylation reprograms BM cells to a cardiac-competent progenitor phenotype.

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Inhibition of GSK3β enhances both adhesive and signalling activities of β‐catenin in mouse embryonic stem cells

Cited by 59 publications

References 35 publications

Insulin Increases Epiblast Cell Number of In Vitro Cultured Mouse Embryos via the PI3K/GSK3/p53 Pathway

Insulin Increases Epiblast Cell Number of In Vitro Cultured Mouse Embryos via the PI3K/GSK3/p53 Pathway

Wnt/β-catenin signaling directs the regional expansion of first and second heart field-derived ventricular cardiomyocytes

The Histone Methyltransferase Inhibitor BIX01294 Enhances the Cardiac Potential of Bone Marrow Cells

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