title: miRNA-1 and miRNA-133a are involved in early commitment of pluripotent stem cells and demonstrate antagonistic roles in the regulation of cardiac differentiation.
abstract: miRNA-1 (miR-1) and miRNA-133a (miR-133a) are muscle-specific miRNAs that play an important role in heart development and physiopathology. Although both miRNAs have been broadly studied during cardiogenesis, the mechanisms by which miR-1 and miR-133a could influence linage commitment in pluripotent stem cells remain poorly characterized. In this study we analysed the regulation of miR-1 and miR-133a expression during pluripotent stem cell differentiation [P19.CL6 cells; embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs)] and investigated their role in DMSO and embryoid body (EB)-mediated mesodermal and cardiac differentiation by gain- and loss-of-function studies, as well as in vivo, by the induction of teratomas. Gene expression analysis revealed that miR-1 and miR-133a are upregulated during cardiac differentiation of P19.CL6 cells, and also during ESC and iPSC EB differentiation. Forced overexpression of both miRNAs promoted mesodermal commitment and a concomitant decrease in the expression of neural differentiation markers. Moreover, overexpression of miR-1 enhanced the cardiac differentiation of P19.CL6, while miR-133a reduced it with respect to control cells. Teratoma formation experiments with P19.CL6 cells confirmed the influence of miR-1 and miR-133a during in vivo differentiation. Finally, inhibition of both miRNAs during P19.CL6 cardiac differentiation had opposite results to their overexpression. In conclusion, gene regulation involving miR-1 and miR-133a controls the mesodermal and cardiac fate of pluripotent stem cells. Copyright © 2014 John Wiley & Sons, Ltd.
authors.affiliation: Department of Cardiovascular Development and Repair, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain.|Immunology and Oncology Department, National Centre for Biotechnology, CSIC, Madrid, Spain.
authors.forename: Alberto
authors.initials: A
authors.lastname: Izarra
authors.affiliation: Department of Cardiovascular Development and Repair, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain.|Cardiovascular Area, CIMUS, Instituto de Investigación Sanitaria, University of Santiago de Compostela, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Moscoso
authors.affiliation: Department of Cardiovascular Development and Repair, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain.|Immunology and Oncology Department, National Centre for Biotechnology, CSIC, Madrid, Spain.
authors.forename: Susana
authors.initials: S
authors.lastname: Cañón
authors.affiliation: Department of Cardiovascular Development and Repair, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain.
authors.forename: Candelas
authors.initials: C
authors.lastname: Carreiro
authors.affiliation: Departament de Medicina i Cirurgia Animals, Universitat Autònoma de Barcelona, Spain.
authors.forename: Dolors
authors.initials: D
authors.lastname: Fondevila
authors.affiliation: Unidad de Animal de Laboratorio, PRBB, Barcelona, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Martín-Caballero
authors.affiliation: Department of Cardiovascular Development and Repair, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Blanca
authors.affiliation: Department of Cardiovascular Development and Repair, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain.
authors.forename: Iñigo
authors.initials: I
authors.lastname: Valiente
authors.affiliation: Department of Cardiovascular Development and Repair, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain.|Instituto de Investigación Sanitaria INCLIVA, Valencia, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Díez-Juan
authors.affiliation: Department of Cardiovascular Development and Repair, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain.|Immunology and Oncology Department, National Centre for Biotechnology, CSIC, Madrid, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Bernad
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D019070:Cell Lineage / Q000235:genetics*; D005786:Gene Expression Regulation; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology; D016513:Mice, SCID; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D009206:Myocardium / Q000166:cytology*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*
chemical_list: D035683:MicroRNAs; C517731:Mirn1 microRNA, mouse; C517732:Mirn133 microRNA, mouse
keywords: P19; cardiac differentiation; embryonic stem cells; induced pluripotent stem cells; microRNAs; pluripotent stem cells