title: Comparative expression analysis of Shox2-deficient embryonic stem cell-derived sinoatrial node-like cells.
abstract: The homeodomain transcription factor Shox2 controls the development and function of the native cardiac pacemaker, the sinoatrial node (SAN). Moreover, SHOX2 mutations have been associated with cardiac arrhythmias in humans. For detailed examination of Shox2-dependent developmental mechanisms in SAN cells, we established a murine embryonic stem cell (ESC)-based model using Shox2 as a molecular tool. Shox2+/+and Shox2-/-ESC clones were isolated and differentiated according to five different protocols in order to evaluate the most efficient enrichment of SAN-like cells. Expression analysis of cell subtype-specific marker genes revealed most efficient enrichment after CD166-based cell sorting. Comparative cardiac expression profiles of Shox2+/+and Shox2-/-ESCs were examined by nCounter technology. Among other genes, we identified Nppb as a novel putative Shox2 target during differentiation in ESCs. Differential expression of Nppb could be confirmed in heart tissue of Shox2-/-embryos. Taken together, we established an ESC-based cardiac differentiation model and successfully purified Shox2+/+and Shox2-/-SAN-like cells. This now provides an excellent basis for the investigation of molecular mechanisms under physiological and pathophysiological conditions for evaluating novel therapeutic approaches.
authors.affiliation: Department of Human Molecular Genetics, Institute of Human Genetics, University Heidelberg, Germany; DZHK, German Centre for Cardiovascular Research, Partner site Heidelberg/Mannheim, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Hoffmann
authors.affiliation: Department of Human Molecular Genetics, Institute of Human Genetics, University Heidelberg, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Schmitteckert
authors.affiliation: Department of Human Molecular Genetics, Institute of Human Genetics, University Heidelberg, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Griesbeck
authors.affiliation: Department of Human Molecular Genetics, Institute of Human Genetics, University Heidelberg, Germany.
authors.forename: Hannes
authors.initials: H
authors.lastname: Preiss
authors.affiliation: Department of Human Molecular Genetics, Institute of Human Genetics, University Heidelberg, Germany; DZHK, German Centre for Cardiovascular Research, Partner site Heidelberg/Mannheim, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Sumer
authors.affiliation: Institute for Biological Interfaces, Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Rolletschek
authors.affiliation: Department of Human Genetics, Institute of Human Genetics, University Heidelberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Granzow
authors.affiliation: FACS Core Facility, Department of Medicine V, University Hospital Heidelberg, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Eckstein
authors.affiliation: Department of Human Molecular Genetics, Institute of Human Genetics, University Heidelberg, Germany; DZHK, German Centre for Cardiovascular Research, Partner site Heidelberg/Mannheim, Germany; nCounter Core Facility, Department of Human Molecular Genetics, Institute of Human Genetics, University Heidelberg, Germany.
authors.forename: Beate
authors.initials: B
authors.lastname: Niesler
authors.affiliation: Department of Human Molecular Genetics, Institute of Human Genetics, University Heidelberg, Germany; DZHK, German Centre for Cardiovascular Research, Partner site Heidelberg/Mannheim, Germany. Electronic address: Gudrun.Rappold@med.uni-heidelberg.de.
authors.forename: Gudrun A
authors.initials: GA
authors.lastname: Rappold
mesh_terms: D020410:Activated-Leukocyte Cell Adhesion Molecule / Q000378:metabolism; D000818:Animals; D002469:Cell Separation; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D012849:Sinoatrial Node / Q000166:cytology*
chemical_list: D020410:Activated-Leukocyte Cell Adhesion Molecule; D018398:Homeodomain Proteins; C484663:Shox2 protein, mouse
keywords: Differentiation model; Mouse embryonic stem cells; Shox2; Sinoatrial node-like cells; Transgenic cells