Myeloid zinc finger 1 (Mzf1) differentially modulates murine cardiogenesis by interacting with an Nkx2.5 cardiac enhancer

Stefanie A Doppler; Astrid Werner; Melanie Barz; Harald Lahm; Marcus-André Deutsch; Martina Dreßen; Matthias Schiemann; Bernhard Voss; Serge Gregoire; Rajarajan Kuppusamy; Sean M Wu; Rüdiger Lange; Markus Krane

doi:10.1371/journal.pone.0113775

Myeloid zinc finger 1 (Mzf1) differentially modulates murine cardiogenesis by interacting with an Nkx2.5 cardiac enhancer

PLoS One. 2014 Dec 1;9(12):e113775. doi: 10.1371/journal.pone.0113775. eCollection 2014.

Affiliations

¹ Department of Experimental Surgery, Department of Cardiovascular Surgery, Deutsches Herzzentrum München, Technische Universität München (TUM), Munich, Germany.
² Institute for Medical Microbiology, Immunology and Hygiene, Technische Universität München (TUM), Munich, Germany; Clinical Cooperation Groups "Antigen-specific Immunotherapy" and "Immune-Monitoring", Helmholtz Center Munich (Neuherberg), TUM, Munich, Germany.
³ Cardiovascular Research Center, Division of Cardiology, Harvard Medical School, Department of Medicine, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
⁴ Division of Cardiovascular Medicine, Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California, United States of America; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, United States of America.
⁵ Department of Experimental Surgery, Department of Cardiovascular Surgery, Deutsches Herzzentrum München, Technische Universität München (TUM), Munich, Germany; DZHK (German Center for Cardiovascular Research) - partner site Munich Heart Alliance, Munich, Germany.

Abstract

Vertebrate heart development is strictly regulated by temporal and spatial expression of growth and transcription factors (TFs). We analyzed nine TFs, selected by in silico analysis of an Nkx2.5 enhancer, for their ability to transactivate the respective enhancer element that drives, specifically, expression of genes in cardiac progenitor cells (CPCs). Mzf1 showed significant activity in reporter assays and bound directly to the Nkx2.5 cardiac enhancer (Nkx2.5 CE) during murine ES cell differentiation. While Mzf1 is established as a hematopoietic TF, its ability to regulate cardiogenesis is completely unknown. Mzf1 expression was significantly enriched in CPCs from in vitro differentiated ES cells and in mouse embryonic hearts. To examine the effect of Mzf1 overexpression on CPC formation, we generated a double transgenic, inducible, tetOMzf1-Nkx2.5 CE eGFP ES line. During in vitro differentiation an early and continuous Mzf1 overexpression inhibited CPC formation and cardiac gene expression. A late Mzf1 overexpression, coincident with a second physiological peak of Mzf1 expression, resulted in enhanced cardiogenesis. These findings implicate a novel, temporal-specific role of Mzf1 in embryonic heart development. Thereby we add another piece of puzzle in understanding the complex mechanisms of vertebrate cardiac development and progenitor cell differentiation. Consequently, this knowledge will be of critical importance to guide efficient cardiac regenerative strategies and to gain further insights into the molecular basis of congenital heart malformations.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation
Cell Line
Cells, Cultured
Computer Simulation
Embryonic Stem Cells / cytology
Enhancer Elements, Genetic*
Gene Expression Regulation, Developmental
HEK293 Cells
Heart / embryology*
Homeobox Protein Nkx-2.5
Homeodomain Proteins / genetics*
Humans
Kruppel-Like Transcription Factors / genetics*
Kruppel-Like Transcription Factors / metabolism*
Mice
Mice, Transgenic
Transcription Factors / genetics*

Substances

Homeobox Protein Nkx-2.5
Homeodomain Proteins
Kruppel-Like Transcription Factors
Mzf1 protein, mouse
Nkx2-5 protein, mouse
Transcription Factors

Grants and funding

This work was supported by the Dr. Rusche Forschungspreis 2011 of the Deutsche Gesellschaft für Thorax-, Herz- und Gefäßchirurgie (http://www.dshf.de/dr_rusche_forschungsprojekt_projekte.php). Grant Support (Doppler et al.): The study was supported by Dr. Rusche Forschungsprojekt (2011) of the DSHF and DGTHG. Marcus-André Deutsch (MAD) is supported by Dr. Rusche Forschungsprojekt (2014) of the DSHF and DGTHG. Rüdiger Lange (RL) is supported by Bayerische Forschungsstiftung (AZ-1012-12). Markus Krane (MK) is supported by Deutsche Stiftung für Herzforschung (F/37/11), Deutsches Zentrum für Herz Kreislauf Forschung (DZHK B 13-050A), Deutsche Forschungsgemeinschaft – Sachmittelantrag (KR3770/7-1), Deutsches Zentrum für Herz Kreislauf Forschung (DZHK B 14-013SE), and Deutsche Forschungsgemeinschaft – Sachmittelantrag (KR3770/9-1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.