TY - JOUR
T1 - Zeb2 regulates cell fate at the exit from epiblast state in mouse embryonic stem cells
AU - Stryjewska, Agata
AU - Dries, Ruben
AU - Pieters, Tim
AU - Verstappen, Griet
AU - Conidi, Andrea
AU - Coddens, Kathleen
AU - Francis, Annick
AU - Umans, Lieve
AU - van Ijcken, Wilfred F. J.
AU - Berx, Geert
AU - van Grunsven, Leo A.
AU - Grosveld, Frank
AU - Goossens, Steven
AU - Haigh, Jody J.
AU - Huylebroeck, Danny
PY - 2017/3
Y1 - 2017/3
N2 - In human ESCs the transcription factor Zeb2 regulates neuroectoderm versus mesendoderm formation, but it is unclear how Zeb2 affects the global transcriptional regulatory network in these cell-fate decisions. We generated Zeb2 knockout (KO) mouse ESCs, subjected them as embryoid bodies (EBs) to neural and general differentiation and carried out temporal RNA-sequencing (RNA-seq) and reduced representation bisulfite sequencing (RRBS) analysis in neural differentiation. This shows that Zeb2 acts preferentially as a transcriptional repressor associated with developmental progression and that Zeb2 KO ESCs can exit from their naïve state. However, most cells in these EBs stall in an early epiblast-like state and are impaired in both neural and mesendodermal differentiation. Genes involved in pluripotency, epithelial-to-mesenchymal transition (EMT) and DNA-(de)methylation, including Tet1, are deregulated in the absence of Zeb2. The observed elevated Tet1 levels in the mutant cells and the knowledge of previously mapped Tet1-binding sites correlate with loss-of-methylation in neural-stimulating conditions, however after the cells initially acquired the correct DNA-methyl marks. Interestingly, cells from such Zeb2 KO EBs maintain the ability to re-adapt to 2i+LIF conditions even after prolonged differentiation, while knockdown of Tet1 partially rescues their impaired differentiation. Hence, in addition to its role in EMT, Zeb2 is critical in ESCs for exit from the epiblast state, and links the pluripotency network and DNA-methylation with irreversible commitment to differentiation. This article is protected by copyright. All rights reserved.
AB - In human ESCs the transcription factor Zeb2 regulates neuroectoderm versus mesendoderm formation, but it is unclear how Zeb2 affects the global transcriptional regulatory network in these cell-fate decisions. We generated Zeb2 knockout (KO) mouse ESCs, subjected them as embryoid bodies (EBs) to neural and general differentiation and carried out temporal RNA-sequencing (RNA-seq) and reduced representation bisulfite sequencing (RRBS) analysis in neural differentiation. This shows that Zeb2 acts preferentially as a transcriptional repressor associated with developmental progression and that Zeb2 KO ESCs can exit from their naïve state. However, most cells in these EBs stall in an early epiblast-like state and are impaired in both neural and mesendodermal differentiation. Genes involved in pluripotency, epithelial-to-mesenchymal transition (EMT) and DNA-(de)methylation, including Tet1, are deregulated in the absence of Zeb2. The observed elevated Tet1 levels in the mutant cells and the knowledge of previously mapped Tet1-binding sites correlate with loss-of-methylation in neural-stimulating conditions, however after the cells initially acquired the correct DNA-methyl marks. Interestingly, cells from such Zeb2 KO EBs maintain the ability to re-adapt to 2i+LIF conditions even after prolonged differentiation, while knockdown of Tet1 partially rescues their impaired differentiation. Hence, in addition to its role in EMT, Zeb2 is critical in ESCs for exit from the epiblast state, and links the pluripotency network and DNA-methylation with irreversible commitment to differentiation. This article is protected by copyright. All rights reserved.
KW - cell differentiation
KW - DNA‐methylation
KW - embryonic stem cells
KW - pluripotent stem cells
KW - repressors
KW - RNA‐sequencing
KW - transcription factors
KW - transcriptom
UR - http://onlinelibrary.wiley.com/doi/10.1002/stem.2521/full
U2 - 10.1002/stem.2521
DO - 10.1002/stem.2521
M3 - Article
SN - 1066-5099
VL - 35
SP - 611
EP - 625
JO - Stem Cells
JF - Stem Cells
IS - 3
ER -