title: Stepwise Clearance of Repressive Roadblocks Drives Cardiac Induction in Human ESCs.
abstract: Cardiac induction requires stepwise integration of BMP and WNT pathway activity. Human embryonic stem cells (hESCs) are developmentally and clinically relevant for studying the poorly understood molecular mechanisms downstream of these cascades. We show that BMP and WNT signaling drive cardiac specification by removing sequential roadblocks that otherwise redirect hESC differentiation toward competing fates, rather than activating a cardiac program per se. First, BMP and WNT signals pattern mesendoderm through cooperative repression of SOX2, a potent mesoderm antagonist. BMP signaling promotes miRNA-877 maturation to induce SOX2 mRNA degradation, while WNT-driven EOMES induction transcriptionally represses SOX2. Following mesoderm formation, cardiac differentiation requires inhibition of WNT activity. We found that WNT inhibition serves to restrict expression of anti-cardiac regulators MSX1 and CDX2/1. Conversely, their simultaneous disruption partially abrogates the requirement for WNT inactivation. These results suggest that human cardiac induction depends on multi-stage repression of alternate lineages, with implications for deriving expandable cardiac stem cells.
authors.affiliation: Human Stem Cell Pluripotency Laboratory, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany; Chemical Genomics Centre of the Max Planck Society, 44227 Dortmund, Germany.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Rao
authors.affiliation: Human Stem Cell Pluripotency Laboratory, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany; Chemical Genomics Centre of the Max Planck Society, 44227 Dortmund, Germany.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Pfeiffer
authors.affiliation: Human Stem Cell Pluripotency Laboratory, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany; Chemical Genomics Centre of the Max Planck Society, 44227 Dortmund, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Frank
authors.affiliation: Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany.
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Adachi
authors.affiliation: Institute of Genetics of Heart Diseases (IfGH), Department of Cardiovascular Medicine, University Hospital Münster, 48149 Münster, Germany.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Piccini
authors.affiliation: Human Stem Cell Pluripotency Laboratory, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany; Chemical Genomics Centre of the Max Planck Society, 44227 Dortmund, Germany.
authors.forename: Roberto
authors.initials: R
authors.lastname: Quaranta
authors.affiliation: Group of Computational Biology and Systems Biomedicine, Biodonostia Health Research Institute, 20014 San Sebastián, Spain; IKERBASQUE, Basque Foundation for Science, 48009 Bilbao, Spain.
authors.forename: Marcos
authors.initials: M
authors.lastname: Araúzo-Bravo
authors.affiliation: Max Planck Research Group for RNA Biology, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany; Cells-in-Motion Cluster of Excellence, Faculty of Medicine, University of Münster, 48149 Münster, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Schwarz
authors.affiliation: Technical University of Dortmund, 44227 Dortmund, Germany.
authors.forename: Dennis
authors.initials: D
authors.lastname: Schade
authors.affiliation: Max Planck Research Group for RNA Biology, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany; Cells-in-Motion Cluster of Excellence, Faculty of Medicine, University of Münster, 48149 Münster, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Leidel
authors.affiliation: Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany.
authors.forename: Hans R
authors.initials: HR
authors.lastname: Schöler
authors.affiliation: Institute of Genetics of Heart Diseases (IfGH), Department of Cardiovascular Medicine, University Hospital Münster, 48149 Münster, Germany.
authors.forename: Guiscard
authors.initials: G
authors.lastname: Seebohm
authors.affiliation: Human Stem Cell Pluripotency Laboratory, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany; Chemical Genomics Centre of the Max Planck Society, 44227 Dortmund, Germany. Electronic address: boris.greber@mpi-muenster.mpg.de.
authors.forename: Boris
authors.initials: B
authors.lastname: Greber
mesh_terms: D000071616:CDX2 Transcription Factor / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D051957:MSX1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D060449:Wnt Signaling Pathway
chemical_list: C491273:CDX1 protein, human; D000071616:CDX2 Transcription Factor; C491270:CDX2 protein, human; C491199:EOMES protein, human; D018398:Homeodomain Proteins; C570298:MIRN877 microRNA, human; D051957:MSX1 Transcription Factor; C484083:MSX1 protein, human; D035683:MicroRNAs; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D020825:T-Box Domain Proteins