title: EBIO Does Not Induce Cardiomyogenesis in Human Pluripotent Stem Cells but Modulates Cardiac Subtype Enrichment by Lineage-Selective Survival.
abstract: Subtype-specific human cardiomyocytes (CMs) are valuable for basic and applied research. Induction of cardiomyogenesis and enrichment of nodal-like CMs was described for mouse pluripotent stem cells (mPSCs) in response to 1-ethyl-2-benzimidazolinone (EBIO), a chemical modulator of small-/intermediate-conductance Ca2+-activated potassium channels (SKs 1-4). Investigating EBIO in human pluripotent stem cells (PSCs), we have applied three independent differentiation protocols of low to high cardiomyogenic efficiency. Equivalent to mPSCs, timed EBIO supplementation during hPSC differentiation resulted in dose-dependent enrichment of up to 80% CMs, including an increase in nodal- and atrial-like phenotypes. However, our study revealed extensive EBIO-triggered cell loss favoring cardiac progenitor preservation and, subsequently, CMs with shortened action potentials. Proliferative cells were generally more sensitive to EBIO, presumably via an SK-independent mechanism. Together, EBIO did not promote cardiogenic differentiation of PSCs, opposing previous findings, but triggered lineage-selective survival at a cardiac progenitor stage, which we propose as a pharmacological strategy to modulate CM subtype composition.
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg Straße 1, 30625 Hannover, Germany.
authors.forename: Monica
authors.initials: M
authors.lastname: Jara-Avaca
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg Straße 1, 30625 Hannover, Germany.
authors.forename: Henning
authors.initials: H
authors.lastname: Kempf
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg Straße 1, 30625 Hannover, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Rückert
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg Straße 1, 30625 Hannover, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Robles-Diaz
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg Straße 1, 30625 Hannover, Germany.
authors.forename: Annika
authors.initials: A
authors.lastname: Franke
authors.affiliation: Institute for Neurophysiology, Hannover Medical School, Carl-Neuberg Straße, 30625 Hannover, Germany.
authors.forename: Jeanne
authors.initials: J
authors.lastname: de la Roche
authors.affiliation: Institute for Neurophysiology, Hannover Medical School, Carl-Neuberg Straße, 30625 Hannover, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Fischer
authors.affiliation: Institute of Physiology I, Life & Brain Center, University of Bonn, Sigmund-Freud-Straße 25, 53127 Bonn, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Malan
authors.affiliation: Institute of Physiology I, Life & Brain Center, University of Bonn, Sigmund-Freud-Straße 25, 53127 Bonn, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Sasse
authors.affiliation: Center of Biomolecular Drug Research (BMWZ), Institute of Organic Chemistry, Leibniz University Hannover, Schneiderberg 38, 30167 Hannover, Germany.
authors.forename: Wladimir
authors.initials: W
authors.lastname: Solodenko
authors.affiliation: Center of Biomolecular Drug Research (BMWZ), Institute of Organic Chemistry, Leibniz University Hannover, Schneiderberg 38, 30167 Hannover, Germany.
authors.forename: Gerald
authors.initials: G
authors.lastname: Dräger
authors.affiliation: Center of Biomolecular Drug Research (BMWZ), Institute of Organic Chemistry, Leibniz University Hannover, Schneiderberg 38, 30167 Hannover, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kirschning
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg Straße 1, 30625 Hannover, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Martin
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg Straße 1, 30625 Hannover, Germany. Electronic address: zweigerdt.robert@mh-hannover.de.
authors.forename: Robert
authors.initials: R
authors.lastname: Zweigerdt
mesh_terms: D001562:Benzimidazoles / Q000494:pharmacology*; D015415:Biomarkers; D002120:Calcium Channel Agonists / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D002460:Cell Line; D019070:Cell Lineage; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels / Q000235:genetics / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology / Q000187:drug effects / Q000196:embryology; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism
chemical_list: D001562:Benzimidazoles; D015415:Biomarkers; D002120:Calcium Channel Agonists; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels; C496923:KCNN4 protein, human; C102635:1-ethyl-2-benzimidazolinone
keywords: 1-ethyl-2-benzimidazolinone (EBIO); CyPPA; NS309; SK channel; cardiomyocyte enrichment; cardiomyocyte subtype; differentiation; human pluripotent stem cells; proliferation