title: Small molecule-mediated directed differentiation of human embryonic stem cells toward ventricular cardiomyocytes.
abstract: The generation of human ventricular cardiomyocytes from human embryonic stem cells and/or induced pluripotent stem cells could fulfill the demand for therapeutic applications and in vitro pharmacological research; however, the production of a homogeneous population of ventricular cardiomyocytes remains a major limitation. By combining small molecules and growth factors, we developed a fully chemically defined, directed differentiation system to generate ventricular-like cardiomyocytes (VCMs) from human embryonic stem cells and induced pluripotent stem cells with high efficiency and reproducibility. Molecular characterization revealed that the differentiation recapitulated the developmental steps of cardiovascular fate specification. Electrophysiological analyses further illustrated the generation of a highly enriched population of VCMs. These chemically induced VCMs exhibited the expected cardiac electrophysiological and calcium handling properties as well as the appropriate chronotropic responses to cardioactive compounds. In addition, using an integrated computational and experimental systems biology approach, we demonstrated that the modulation of the canonical Wnt pathway by the small molecule IWR-1 plays a key role in cardiomyocyte subtype specification. In summary, we developed a reproducible and efficient experimental platform that facilitates a chemical genetics-based interrogation of signaling pathways during cardiogenesis that bypasses the limitations of genetic approaches and provides a valuable source of ventricular cardiomyocytes for pharmacological screenings as well as cell replacement therapies.
authors: Cardiovascular Research Center and Department of Pharmacology and Systems Therapeutics, Systems Biology Center, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Stem Cell and Regenerative Medicine Consortium, Department of Physiology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong.
authors: Ioannis
authors: I
authors: Karakikes
authors: Grant D
authors: GD
authors: Senyei
authors: Jens
authors: J
authors: Hansen
authors: Chi-Wing
authors: CW
authors: Kong
authors: Evren U
authors: EU
authors: Azeloglu
authors: Francesca
authors: F
authors: Stillitano
authors: Deborah K
authors: DK
authors: Lieu
authors: Jiaxian
authors: J
authors: Wang
authors: Lihuan
authors: L
authors: Ren
authors: Jean-Sebastien
authors: JS
authors: Hulot
authors: Ravi
authors: R
authors: Iyengar
authors: Ronald A
authors: RA
authors: Li
authors: Roger J
authors: RJ
authors: Hajjar
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D028341:Activins / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D003470:Culture Media / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D006352:Heart Ventricles / Q000166:cytology*; D006801:Humans; D007094:Imides / Q000494:pharmacology*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology; D011804:Quinolines / Q000494:pharmacology*; D059467:Transcriptome / Q000187:drug effects / Q000502:physiology; D060449:Wnt Signaling Pathway / Q000187:drug effects / Q000502:physiology
chemical_list: D000970:Antineoplastic Agents; C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D003470:Culture Media; C585534:IWR-1 compound; D007094:Imides; D011804:Quinolines; C422405:activin A; D028341:Activins; D002118:Calcium
keywords: Cardiac; Differentiation; Embryonic stem cells; Pluripotent stem cells