title: BMP-SMAD Signaling Regulates Lineage Priming, but Is Dispensable for Self-Renewal in Mouse Embryonic Stem Cells.
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.
authors.affiliation: Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
authors.forename: Maria
authors.initials: M
authors.lastname: Gomes Fernandes
authors.affiliation: 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.
authors.forename: Ruben
authors.initials: R
authors.lastname: Dries
authors.affiliation: Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
authors.forename: Matthias S
authors.initials: MS
authors.lastname: Roost
authors.affiliation: Leiden Institute of Physics, Leiden University, Leiden 2333 CA, the Netherlands.
authors.forename: Stefan
authors.initials: S
authors.lastname: Semrau
authors.affiliation: Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
authors.forename: Ana
authors.initials: A
authors.lastname: de Melo Bernardo
authors.affiliation: Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
authors.forename: Richard P
authors.initials: RP
authors.lastname: Davis
authors.affiliation: Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
authors.forename: Ramprasad
authors.initials: R
authors.lastname: Ramakrishnan
authors.affiliation: Department Molecular Cell Biology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
authors.forename: Karoly
authors.initials: K
authors.lastname: Szuhai
authors.affiliation: Department Human Genetics, VIB Center for the Biology of Disease, KU Leuven, Leuven 3000, Belgium.
authors.forename: Elke
authors.initials: E
authors.lastname: Maas
authors.affiliation: 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.
authors.forename: Lieve
authors.initials: L
authors.lastname: Umans
authors.affiliation: Department Human Genetics, VIB Center for the Biology of Disease, KU Leuven, Leuven 3000, Belgium.
authors.forename: Vanesa
authors.initials: V
authors.lastname: Abon Escalona
authors.affiliation: 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.
authors.forename: Daniela
authors.initials: D
authors.lastname: Salvatori
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Ghent 9000, Belgium.
authors.forename: Dieter
authors.initials: D
authors.lastname: Deforce
authors.affiliation: Mathematical Modelling, Statistics and Bio-informatics, Faculty Bioscience Engineering, Ghent University, Ghent 9000, Belgium.
authors.forename: Wim
authors.initials: W
authors.lastname: Van Criekinge
authors.affiliation: 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.
authors.forename: Danny
authors.initials: D
authors.lastname: Huylebroeck
authors.affiliation: Department Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, the Netherlands.
authors.forename: Christine
authors.initials: C
authors.lastname: Mummery
authors.affiliation: Department Human Genetics, VIB Center for the Biology of Disease, KU Leuven, Leuven 3000, Belgium.
authors.forename: An
authors.initials: A
authors.lastname: Zwijsen
authors.affiliation: 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.
authors.forename: Susana M Chuva
authors.initials: SM
authors.lastname: de Sousa Lopes
mesh_terms: D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000235:genetics / Q000378:metabolism*; D019070:Cell Lineage / Q000235:genetics / Q000502:physiology*; D000066673:Cell Self Renewal / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D049308:Dioxygenases; D020869:Gene Expression Profiling / Q000379:methods; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology*; D051896:Smad Proteins, Receptor-Regulated / Q000235:genetics / Q000378:metabolism*; D051898:Smad1 Protein / Q000235:genetics / Q000378:metabolism; D051902:Smad5 Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D019485:Bone Morphogenetic Proteins; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D051896:Smad Proteins, Receptor-Regulated; D051898:Smad1 Protein; D051902:Smad5 Protein; C540839:TET1 protein, mouse; D049308:Dioxygenases; C551914:Tet2 protein, mouse; D004248:DNA (Cytosine-5-)-Methyltransferases