Cell cycle dynamics of mouse embryonic stem cells in the ground state and during transition to formative pluripotency

Waisman, Ariel; Sevlever, Federico; Costa, Martín Elías; Cosentino, María Soledad; Miriuka, Santiago Gabriel; Ventura, Alejandra C.; Guberman, Alejandra

doi:10.1038/s41598-019-44537-0

Cited by 42 publications

(44 citation statements)

References 37 publications

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“…3C) (p-value = 1.108x10 -8 ). Therefore shortening of the cell-cycle begins prior to exit from naïve pluripotency, as recently proposed(Waisman et al, 2019), and the cell cycle shortens further over subsequent generations(Fig. 3D).…”

supporting

confidence: 68%

“…Generation times reveal that cell division rate also begins to shorten prior to exit from naïve pluripotency. Cell cycle shortening may therefore be an early marker of differentiation , as suggested in a recent report (Waisman et al, 2019) . However, self-renewing ES cells in serum/LIF exhibit a faster cell cycle than 2i cells, with a shortened G1 phase (ter Huurne et al, 2017) .…”

Section: Discussionmentioning

confidence: 70%

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Embryonic stem cells commit to differentiation by symmetric divisions following a variable lag period

Strawbridge

Blanchard

Smith

et al. 2020

Preprint

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Mouse embryonic stem (ES) cells are derived from the epiblast of the preimplantation embryo and retain the capacity to give rise to all embryo lineages. ES cells can be released into differentiation from a near-homogeneous maintenance condition. Exit from the ES cell state can be accurately monitored using the Rex1-GFPd2 transgenic reporter, providing a powerful system for examining a mammalian cell fate transition. Here, we performed live-cell imaging and tracking of ES cells during entry into differentiation for 48 hours in defined conditions. We observed a greater cell surface area and a modest shortening of the cell cycle prior to exit and subsequently a reduction in cell size and increase in motility. We did not see any instance of cells regaining ES cell identity, consistent with unidirectional developmental progression. Transition occurred asynchronously across the population but genealogical tracking revealed a high correlation in cell-cycle length and Rex1-GFPd2 expression between daughter cells. A population dynamics model was consistent with symmetric divisions during exit from naive pluripotency. Collapse of ES cell identity occurred acutely in individual cells but after a variable delay. The variation in lag period can extend up to three generations, creating marked population asynchrony.2

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supporting

confidence: 68%

Section: Discussionmentioning

confidence: 70%

Embryonic stem cells commit to differentiation by symmetric divisions following a variable lag period

Strawbridge

Blanchard

Smith

et al. 2020

Preprint

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“…5E and 5F). This is in agreement with previous reports that indicated that the G2/M state is more associated with pluripotency than the G1 state 54 .…”

Section: During Cellular Differentiation Linage Outcomes Are Not Affsupporting

confidence: 94%

Esrrbis a cell cycle dependent XEN priming factor balancing between pluripotency and differentiation

Levi

Arnon

Feldman

et al. 2020

Preprint

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Cell cycle and differentiation decisions are tightly linked; however, the underlying principles that drive these decisions are not fully understood. Here, we combined cell-cycle reporter system and single-cell RNA-seq profiling to study the transcriptomes of mouse embryonic stem cells (ESCs) in the context of cell cycle states and differentiation. By applying a retinoic acid-based protocol as a differentiation assay representing exit from pluripotency, we show that Esrrb, a key pluripotent factor is upregulated during G2/M phase and promptly downregulated during exit from pluripotency. Enhancer chromatin states and Esrrb ChIP-seq map expose Esrrb's association with differentiation genes, were in the context of retinoic acid, enhancers associated with differentiation program driving extraembryonic endoderm cells (XENs). We show that only G2/M ESCs were capable of differentiating into XENs, whereas cells in the G1 phase predominantly produce epiblast stem cells (EpiSCs). Furthermore, Cells engineered to overexpress Esrrb allowed G1 ESCs to produce XENs, while Esrrb knockout ESCs completely lost their potential to differentiate into XEN. Interestingly, this phenomenon is unique to the pluripotency state as sorting cells after initiation of differentiation resulted in a cell cycle-independent differentiation decisions. Cells in both G1 and G2/M phases contributed equally to EpiSC and XEN cellular lineages. Taken together, this study reveals an important functional link between Esrrb and cell-cycle states during exit from pluripotency. Our novel approach of applying single cell RNAseq in a cell cycle dependent manner can be further expand into other differentiation assays and expand our understanding of early differentiation events.

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“…Taking a step beyond, we proved that mESCs can memorize morphine exposure and maintain reduced levels of H3K27me3 for a minimum of 4 cell cycles. Considering that mESCs have a cell cycle length of approximately 11–14 h [ 56 ], morphine reduces H3K27me3 levels that persist at least 48 h after treatment removal before recovering to control values. In line with this, transcriptional downregulation of some PRC2 components also persists for 48 h, which implies 4 cell cycles, but not 96 h. While elevated levels of promoter H3K27me3 may well be involved in the initial morphine-induced downregulation of PRC2 genes in the short term, it does not seem necessary for the persistence of this effect through multiple cell divisions.…”

Section: Discussionmentioning

confidence: 99%

Morphine leads to global genome changes in H3K27me3 levels via a Polycomb Repressive Complex 2 (PRC2) self-regulatory mechanism in mESCs

et al. 2020

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Background Environmentally induced epigenetic changes can lead to health problems or disease, but the mechanisms involved remain unclear. Morphine can pass through the placental barrier leading to abnormal embryo development. However, the mechanism by which morphine causes these effects and how they sometimes persist into adulthood is not well known. To unravel the morphine-induced chromatin alterations involved in aberrant embryo development, we explored the role of the H3K27me3/PRC2 repressive complex in gene expression and its transmission across cellular generations in response to morphine. Results Using mouse embryonic stem cells as a model system, we found that chronic morphine treatment induces a global downregulation of the histone modification H3K27me3. Conversely, ChIP-Seq showed a remarkable increase in H3K27me3 levels at specific genomic sites, particularly promoters, disrupting selective target genes related to embryo development, cell cycle and metabolism. Through a self-regulatory mechanism, morphine downregulated the transcription of PRC2 components responsible for H3K27me3 by enriching high H3K27me3 levels at the promoter region. Downregulation of PRC2 components persisted for at least 48 h (4 cell cycles) following morphine removal, though promoter H3K27me3 levels returned to control levels. Conclusions Morphine induces targeting of the PRC2 complex to selected promoters, including those of PRC2 components, leading to characteristic changes in gene expression and a global reduction in H3K27me3. Following morphine removal, enhanced promoter H3K27me3 levels revert to normal sooner than global H3K27me3 or PRC2 component transcript levels. We suggest that H3K27me3 is involved in initiating morphine-induced changes in gene expression, but not in their maintenance. Graphic abstract Model of Polycomb repressive complex 2 (PRC2) and H3K27me3 alterations induced by chronic morphine exposure. Morphine induces H3K27me3 enrichment at promoters of genes encoding core members of the PRC2 complex and is associated with their transcriptional downregulation.

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Cell cycle dynamics of mouse embryonic stem cells in the ground state and during transition to formative pluripotency

Cited by 42 publications

References 37 publications

Embryonic stem cells commit to differentiation by symmetric divisions following a variable lag period

Embryonic stem cells commit to differentiation by symmetric divisions following a variable lag period

Esrrbis a cell cycle dependent XEN priming factor balancing between pluripotency and differentiation

Morphine leads to global genome changes in H3K27me3 levels via a Polycomb Repressive Complex 2 (PRC2) self-regulatory mechanism in mESCs

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