BMP-SMAD Signaling Regulates Lineage Priming, but Is Dispensable for Self-Renewal in Mouse Embryonic Stem Cells

Maria Gomes Fernandes; Ruben Dries; Matthias S Roost; Stefan Semrau; Ana de Melo Bernardo; Richard P Davis; Ramprasad Ramakrishnan; Karoly Szuhai; Elke Maas; Lieve Umans; Vanesa Abon Escalona; Daniela Salvatori; Dieter Deforce; Wim Van Criekinge; Danny Huylebroeck; Christine Mummery; An Zwijsen; Susana M Chuva de Sousa Lopes

doi:10.1016/j.stemcr.2015.11.012

BMP-SMAD Signaling Regulates Lineage Priming, but Is Dispensable for Self-Renewal in Mouse Embryonic Stem Cells

Stem Cell Reports. 2016 Jan 12;6(1):85-94. doi: 10.1016/j.stemcr.2015.11.012. Epub 2015 Dec 17.

Authors

Maria Gomes Fernandes¹, Ruben Dries², Matthias S Roost¹, Stefan Semrau³, Ana de Melo Bernardo¹, Richard P Davis¹, Ramprasad Ramakrishnan¹, Karoly Szuhai⁴, Elke Maas⁵, Lieve Umans⁶, Vanesa Abon Escalona⁵, Daniela Salvatori⁷, Dieter Deforce⁸, Wim Van Criekinge⁹, Danny Huylebroeck², Christine Mummery¹, An Zwijsen⁵, Susana M Chuva de Sousa Lopes¹⁰

Affiliations

¹ Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
² Department Development and Regeneration, Laboratory of Molecular Biology (Celgen), KU Leuven, Leuven 3000, Belgium; Department of Cell Biology, Erasmus University Medical Center, Rotterdam 3015 CN, the Netherlands.
³ Leiden Institute of Physics, Leiden University, Leiden 2333 CA, the Netherlands.
⁴ Department Molecular Cell Biology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
⁵ Department Human Genetics, VIB Center for the Biology of Disease, KU Leuven, Leuven 3000, Belgium.
⁶ Department Development and Regeneration, Laboratory of Molecular Biology (Celgen), KU Leuven, Leuven 3000, Belgium; Department of Cell Biology, Erasmus University Medical Center, Rotterdam 3015 CN, the Netherlands; Department Human Genetics, VIB Center for the Biology of Disease, KU Leuven, Leuven 3000, Belgium.
⁷ Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands; Center Laboratory Animal Facility, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
⁸ Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Ghent 9000, Belgium.
⁹ Mathematical Modelling, Statistics and Bio-informatics, Faculty Bioscience Engineering, Ghent University, Ghent 9000, Belgium.
¹⁰ Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands; Department Reproductive Medicine, Ghent University Hospital, Ghent 9000, Belgium. Electronic address: lopes@lumc.nl.

Abstract

Naive mouse embryonic stem cells (mESCs) are in a metastable state and fluctuate between inner cell mass- and epiblast-like phenotypes. Here, we show transient activation of the BMP-SMAD signaling pathway in mESCs containing a BMP-SMAD responsive reporter transgene. Activation of the BMP-SMAD reporter transgene in naive mESCs correlated with lower levels of genomic DNA methylation, high expression of 5-methylcytosine hydroxylases Tet1/2 and low levels of DNA methyltransferases Dnmt3a/b. Moreover, naive mESCs, in which the BMP-SMAD reporter transgene was activated, showed higher resistance to differentiation. Using double Smad1;Smad5 knockout mESCs, we showed that BMP-SMAD signaling is dispensable for self-renewal in both naive and ground state. These mutant mESCs were still pluripotent, but they exhibited higher levels of DNA methylation than their wild-type counterparts and had a higher propensity to differentiate. We showed that BMP-SMAD signaling modulates lineage priming in mESCs, by transiently regulating the enzymatic machinery responsible for DNA methylation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Bone Morphogenetic Proteins / genetics
Bone Morphogenetic Proteins / metabolism*
Cell Lineage / genetics
Cell Lineage / physiology*
Cell Self Renewal / genetics
Cell Self Renewal / physiology*
Cells, Cultured
DNA (Cytosine-5-)-Methyltransferases / genetics
DNA (Cytosine-5-)-Methyltransferases / metabolism
DNA Methylation / genetics
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Dioxygenases
Gene Expression Profiling / methods
Mice
Mice, Knockout
Mice, Transgenic
Microscopy, Confocal
Mouse Embryonic Stem Cells / cytology
Mouse Embryonic Stem Cells / metabolism*
Pluripotent Stem Cells / cytology
Pluripotent Stem Cells / metabolism
Proto-Oncogene Proteins / genetics
Proto-Oncogene Proteins / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Signal Transduction / genetics
Signal Transduction / physiology*
Smad Proteins, Receptor-Regulated / genetics
Smad Proteins, Receptor-Regulated / metabolism*
Smad1 Protein / genetics
Smad1 Protein / metabolism
Smad5 Protein / genetics
Smad5 Protein / metabolism

Substances

Bone Morphogenetic Proteins
DNA-Binding Proteins
Proto-Oncogene Proteins
Smad Proteins, Receptor-Regulated
Smad1 Protein
Smad5 Protein
TET1 protein, mouse
Dioxygenases
Tet2 protein, mouse
DNA (Cytosine-5-)-Methyltransferases