Critical role for P53 in regulating the cell cycle of ground state embryonic stem cells

AbstractMouse Embryonic Stem Cells (ESCs) grown in serum-supplemented conditions are characterized by an extremely short G1-phase due to the lack of G1-phase control. Concordantly, the G1-phase-specific P53-P21 pathway is compromised in serum ESCs. Here we provide evidence that P53 is activated upon transition of serum ESCs to their pluripotent ground state using serum-free 2i conditions and modulates G1-phase progression. Our data shows that the elongated G1-phase characteristic of ground state ESCs is dependent on P53. RNA-seq and ChIP-seq analyses reveal that P53 directly regulates the expression of the Retinoblastoma (RB) protein and that the hypo-phosphorylated, active RB protein plays a key role in G1-phase control. Our findings suggest that the P53-P21 pathway is active in ground state 2i ESCs and that its role in the G1-checkpoint is abolished in serum ESCs. Taken together, the data reveals a mechanism by which inactivation of P53 can lead to loss of RB and uncontrolled cell proliferation.

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Critical Role for P53 in Regulating the Cell Cycle of Ground State Embryonic Stem Cells

Stem Cell Reports ◽

10.1016/j.stemcr.2020.01.001 ◽

2020 ◽

Vol 14 (2) ◽

pp. 175-183 ◽

Cited By ~ 3

Author(s):

Menno ter Huurne ◽

Tianran Peng ◽

Guoqiang Yi ◽

Guido van Mierlo ◽

Hendrik Marks ◽

...

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Embryonic Stem Cells ◽

Ground State ◽

Critical Role ◽

Embryonic Stem

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Identification of New Transcription Factors that Can Promote Pluripotent Reprogramming

Stem Cell Reviews and Reports ◽

10.1007/s12015-021-10220-z ◽

2021 ◽

Author(s):

Ping Huang ◽

Jieying Zhu ◽

Yu Liu ◽

Guihuan Liu ◽

Ran Zhang ◽

...

Keyword(s):

Stem Cells ◽

Transcription Factors ◽

Embryonic Stem Cells ◽

Rna Sequencing ◽

Human Urine ◽

Embryonic Stem ◽

Rna Seq ◽

Reprogramming Efficiency ◽

Yamanaka Factors ◽

Urine Cells

Abstract Background Four transcription factors, Oct4, Sox2, Klf4, and c-Myc (the Yamanka factors), can reprogram somatic cells to induced pluripotent stem cells (iPSCs). Many studies have provided a number of alternative combinations to the non-Yamanaka factors. However, it is clear that many additional transcription factors that can generate iPSCs remain to be discovered. Methods The chromatin accessibility and transcriptional level of human embryonic stem cells and human urine cells were compared by Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) and RNA sequencing (RNA-seq) to identify potential reprogramming factors. Selected transcription factors were employed to reprogram urine cells, and the reprogramming efficiency was measured. Urine-derived iPSCs were detected for pluripotency by Immunofluorescence, quantitative polymerase chain reaction, RNA sequencing and teratoma formation test. Finally, we assessed the differentiation potential of the new iPSCs to cardiomyocytes in vitro. Results ATAC-seq and RNA-seq datasets predicted TEAD2, TEAD4 and ZIC3 as potential factors involved in urine cell reprogramming. Transfection of TEAD2, TEAD4 and ZIC3 (in the presence of Yamanaka factors) significantly improved the reprogramming efficiency of urine cells. We confirmed that the newly generated iPSCs possessed pluripotency characteristics similar to normal H1 embryonic stem cells. We also confirmed that the new iPSCs could differentiate to functional cardiomyocytes. Conclusions In conclusion, TEAD2, TEAD4 and ZIC3 can increase the efficiency of reprogramming human urine cells into iPSCs, and provides a new stem cell sources for the clinical application and modeling of cardiovascular disease. Graphical abstract

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Pooled CRISPR-activation screening coupled with single-cell RNA-seq in mouse embryonic stem cells

STAR Protocols ◽

10.1016/j.xpro.2021.100426 ◽

2021 ◽

Vol 2 (2) ◽

pp. 100426

Author(s):

Celia Alda-Catalinas ◽

Melanie A. Eckersley-Maslin ◽

Wolf Reik

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Single Cell ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cells ◽

Rna Seq ◽

Crispr Activation

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Monocytic cells derived from human embryonic stem cells and fetal liver share common differentiation pathways and homeostatic functions

Blood ◽

10.1182/blood-2010-07-295246 ◽

2011 ◽

Vol 117 (11) ◽

pp. 3065-3075 ◽

Cited By ~ 36

Author(s):

Olena Klimchenko ◽

Antonio Di Stefano ◽

Birgit Geoerger ◽

Sofiane Hamidi ◽

Paule Opolon ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Fetal Liver ◽

Critical Role ◽

Embryonic Stem ◽

Blood Monocytes ◽

Monocytic Cell ◽

Monocytic Cells

Abstract The early emergence of macrophages and their large pattern of tissue distribution during development suggest that they may play a critical role in the initial steps of embryogenesis. In the present study, we show that monocytic cells derived from human embryonic stem cells (hESCs) and from fetal liver follow a differentiation pathway different to that of adult cells, leading to specific functions. Embryonic and fetal monocytic cells differentiated from a CD14lowCD16− precursor to form CD14highCD16+ cells without producing the CD14highCD16− cell population that predominates in adult peripheral blood. Both demonstrated an enhanced expression of genes encoding tissue-degrading enzymes, chemokines, and scavenger receptors, as was previously reported for M2 macrophages. Compared with adult blood monocytes, embryonic and fetal monocytic cells secreted high amounts of proteins acting on tissue remodeling and angiogenesis, and most of them expressed the Tie2 receptor. Furthermore, they promoted vascular remodeling in xenotransplanted human tumors. These findings suggest that the regulation of human fetal and embryonic monocytic cell differentiation leads to the generation of cells endowed mainly with anti-inflammatory and remodeling functions. Trophic and immunosuppressive functions of M2-polarized macrophages link fetus and tumor development, and hESCs offer a valuable experimental model for in vitro studies of mechanisms sustaining these processes.

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Abstract 361: Neuregulin Transcriptional Programming of Embryonic Endothelial Progenitor Cells and Embryonic Stem Cells

Circulation Research ◽

10.1161/res.111.suppl_1.a361 ◽

2012 ◽

Vol 111 (suppl_1) ◽

Author(s):

Jijun Hao ◽

Cristi L Galindo ◽

Radwan N Safa ◽

Truc-Linh Tran ◽

Douglas B Sawyer

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Heart Development ◽

Progenitor Cell ◽

Critical Role ◽

Embryonic Stem ◽

Mapk Signaling ◽

P Value ◽

Type I ◽

Endothelial Progenitor

Jijun Hao, Cristi L. Galindo, Radwan N. Safa, Truc-Linh Tran, Douglas B. Sawyer Neuregulin-1 (NRG-1) plays a critical role in heart development by signaling through type I receptor tyrosine kinases in the erbB family (erbB2, erbB3 and erbB4). Mice with disrupted expression of NRG-1, ErbB2, ErbB3 or ErbB4 die in utero with failure of cardiac development. We have previously shown that NRG-1 has distinct effects on two embryonic progenitor cell populations that express ErbB2 and ErbB3 receptors. In an embryonic endothelial progenitor cell line (eEPCs) NRG-1 treatment induces phosphorylation of Akt, GSK-3β, and Erk1/2, and protects eEPCs against serum deprivation-induced apoptosis. In embryonic stem cells (ESCs) we find that NRG-1 treatment from day 0∼2 induces cardiomyocyte formation by day 8 in culture, and when ErbB3 is knocked down in the ESCs, NRG-1 fails to promote cardiomyogenesis. To understand early molecular events that might regulate these distinct effects, we analyzed global transcriptional changes induced by NRG-1 in both eEPCs and ESCs using microarrays. There were only 244 significantly differential (p value < 0.05, fold-change > 1.5) genes detected in NRG-1-treated ESCs, while NRG-1 induced differential expression of 1,547 transcripts in eEPCs. Based on functional analysis, the most significantly over-represented function (Fishers Exact Test, p value with FDR < 0.05) in ESCs was “cell morphogenesis during differentiation”. In eEPCs, genes regulated via Ras/MAPK signaling were altered, as were those downstream of the Akt-PI3K pathway and calcium signaling. For both cell lines, the most statistically significant transcription factor identified as a regulator of the genes altered in response to NRG-1 was SRF, consistent with a role for NRG-1 in heart development and regeneration. Based on the results of this study, we constructed a putative signaling pathway whereby NRG mediates cardiomyogenesis in pluripotent stem cells that correlates with phenotypic observations.

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Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium

Embryonic Stem Cell Protocols - Methods in Molecular Biology ◽

10.1007/7651_2015_207 ◽

2015 ◽

pp. 41-48 ◽

Cited By ~ 7

Author(s):

Matteo Tosolini ◽

Alice Jouneau

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Ground State ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cells

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A Critical Role of Mitochondrial Phosphatase Ptpmt1 in Embryogenesis Reveals a Mitochondrial Metabolic Stress-Induced Differentiation Checkpoint in Embryonic Stem Cells

Molecular and Cellular Biology ◽

10.1128/mcb.05629-11 ◽

2011 ◽

Vol 31 (24) ◽

pp. 4902-4916 ◽

Cited By ~ 23

Author(s):

J. Shen ◽

X. Liu ◽

W.-M. Yu ◽

J. Liu ◽

M. Groot Nibbelink ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Critical Role ◽

Embryonic Stem ◽

Metabolic Stress ◽

Induced Differentiation

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Hippo-YAP signaling controls lineage differentiation of mouse embryonic stem cells through modulating the formation of super-enhancers

Nucleic Acids Research ◽

10.1093/nar/gkaa482 ◽

2020 ◽

Cited By ~ 1

Author(s):

Xiang Sun ◽

Zhijun Ren ◽

Yixian Cun ◽

Cai Zhao ◽

Xianglin Huang ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Signaling Pathway ◽

Embryoid Body ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cells ◽

Hippo Signaling ◽

Rna Seq ◽

Lineage Differentiation ◽

Super Enhancer

Abstract Hippo-YAP signaling pathway functions in early lineage differentiation of pluripotent stem cells, but the detailed mechanisms remain elusive. We found that knockout (KO) of Mst1 and Mst2, two key components of the Hippo signaling in mouse embryonic stem cells (ESCs), resulted in a disruption of differentiation into mesendoderm lineage. To further uncover the underlying regulatory mechanisms, we performed a series of ChIP-seq experiments with antibodies against YAP, ESC master transcription factors and some characterized histone modification markers as well as RNA-seq assays using wild type and Mst KO samples at ES and day 4 embryoid body stage respectively. We demonstrate that YAP is preferentially co-localized with super-enhancer (SE) markers such as Nanog, Sox2, Oct4 and H3K27ac in ESCs. The hyper-activation of nuclear YAP in Mst KO ESCs facilitates the binding of Nanog, Sox2 and Oct4 as well as H3K27ac modification at the loci where YAP binds. Moreover, Mst depletion results in novel SE formation and enhanced liquid-liquid phase-separated Med1 condensates on lineage associated genes, leading to the upregulation of these genes and the distortion of ESC differentiation. Our study reveals a novel mechanism on how Hippo-YAP signaling pathway dictates ESC lineage differentiation.

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