Nkx2-5 Represses Gata1 Gene Expression and Modulates the Cellular Fate of Cardiac Progenitors During Embryogenesis

Background— Recent studies suggest that the hematopoietic and cardiac lineages have close ontogenic origins, and that an early mesodermal cell population has the potential to differentiate into both lineages. Studies also suggest that specification of these lineages is inversely regulated. However, the transcriptional networks that govern the cell fate specification of these progenitors are incompletely defined. Methods and Results— Here, we show that Nkx2-5 regulates the hematopoietic/erythroid fate of the mesoderm precursors early during cardiac morphogenesis. Using transgenic technologies to isolate Nkx2-5 expressing cells, we observed an induction of the erythroid molecular program, including Gata1, in the Nkx2-5–null embryos. We further observed that overexpression of Nkx2-5 with an Nkx2-5–inducible embryonic stem cell system significantly repressed Gata1 gene expression and suppressed the hematopoietic/erythroid potential, but not the endothelial potential, of the embryonic stem cells. This suppression was cell-autonomous, and was partially rescued by overexpressing Gata1. In addition, we demonstrated that Nkx2-5 binds to the Gata1 gene enhancer and represses the transcriptional activity of the Gata1 gene. Conclusions— Our results demonstrate that the hematopoietic/erythroid cell fate is suppressed via Nkx2-5 during mesodermal fate determination, and that the Gata1 gene is one of the targets that are suppressed by Nkx2-5.

[1]  D. Garry,et al.  Myogenic regulatory factors transactivate the Tceal7 gene and modulate muscle differentiation. , 2010, The Biochemical journal.

[2]  Masayuki Yamamoto,et al.  GATA factor switching during erythroid differentiation , 2010, Current opinion in hematology.

[3]  Y. Kaneda,et al.  A histone H3 lysine 36 trimethyltransferase links Nkx2-5 to Wolf–Hirschhorn syndrome , 2009, Nature.

[4]  M. Kyba,et al.  A conserved role for Hox paralog group 4 in regulation of hematopoietic progenitors. , 2009, Stem cells and development.

[5]  R. Hammer,et al.  Nkx2–5 transactivates the Ets-related protein 71 gene and specifies an endothelial/endocardial fate in the developing embryo , 2009, Proceedings of the National Academy of Sciences.

[6]  G. Daley,et al.  Mesodermal patterning activity of SCL. , 2008, Experimental hematology.

[7]  M. Kyba,et al.  ER71 acts downstream of BMP, Notch, and Wnt signaling in blood and vessel progenitor specification. , 2008, Cell stem cell.

[8]  D. Yelon,et al.  Vessel and blood specification override cardiac potential in anterior mesoderm. , 2007, Developmental cell.

[9]  C. Lowrey,et al.  Expression of GATA-1 in a non-hematopoietic cell line induces beta-globin locus control region chromatin structure remodeling and an erythroid pattern of gene expression. , 2007, Journal of molecular biology.

[10]  D. Clapham,et al.  In Brief , 2006, Nature Reviews Drug Discovery.

[11]  E. Olson,et al.  A Common Progenitor at the Heart of Development , 2006, Cell.

[12]  S. Kattman,et al.  Multipotent flk-1+ cardiovascular progenitor cells give rise to the cardiomyocyte, endothelial, and vascular smooth muscle lineages. , 2006, Developmental cell.

[13]  H. Harigae.  GATA transcription factors and hematological diseases. , 2006, The Tohoku journal of experimental medicine.

[14]  Gordon Keller,et al.  Sequential development of hematopoietic and cardiac mesoderm during embryonic stem cell differentiation. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[15]  G. Keller,et al.  Embryonic stem cell differentiation: emergence of a new era in biology and medicine. , 2005, Genes & development.

[16]  Daniel J Garry,et al.  Transcriptional Regulation of Cardiac Progenitor Cell Populations , 2004, Circulation research.

[17]  M. Lotze,et al.  Second‐generation tetracycline‐regulatable promoter: repositioned tet operator elements optimize transactivator synergy while shorter minimal promoter offers tight basal leakiness , 2004, The journal of gene medicine.

[18]  W. Giles,et al.  Nkx2-5 Pathways and Congenital Heart Disease Loss of Ventricular Myocyte Lineage Specification Leads to Progressive Cardiomyopathy and Complete Heart Block , 2004, Cell.

[19]  T. Rabbitts,et al.  Lmo2 and Scl/Tal1 convert non-axial mesoderm into haemangioblasts which differentiate into endothelial cells in the absence of Gata1 , 2003, Development.

[20]  G. Cossu,et al.  Mesoangioblasts--vascular progenitors for extravascular mesodermal tissues. , 2003, Current opinion in genetics & development.

[21]  J. D. Engel,et al.  A minigene containing four discrete cis elements recapitulates GATA‐1 gene expression in vivo , 2002, Genes to cells : devoted to molecular & cellular mechanisms.

[22]  J. McGhee,et al.  The GATA family (vertebrates and invertebrates). , 2002, Current opinion in genetics & development.

[23]  Y. Hiroi,et al.  Csx/Nkx2-5 Is Required for Homeostasis and Survival of Cardiac Myocytes in the Adult Heart* , 2002, The Journal of Biological Chemistry.

[24]  Peter W Zandstra,et al.  Efficiency of embryoid body formation and hematopoietic development from embryonic stem cells in different culture systems. , 2002, Biotechnology and bioengineering.

[25]  S. Orkin,et al.  The search for the hemangioblast. , 2002, Journal of hematotherapy & stem cell research.

[26]  David Baltimore,et al.  Germline Transmission and Tissue-Specific Expression of Transgenes Delivered by Lentiviral Vectors , 2002, Science.

[27]  Y Fujiwara,et al.  Different sequence requirements for expression in erythroid and megakaryocytic cells within a regulatory element upstream of the GATA-1 gene. , 1999, Development.

[28]  S. Izumo,et al.  The cardiac homeobox gene Csx/Nkx2.5 lies genetically upstream of multiple genes essential for heart development. , 1999, Development.

[29]  S. Nishikawa,et al.  Progressive lineage analysis by cell sorting and culture identifies FLK1+VE-cadherin+ cells at a diverging point of endothelial and hemopoietic lineages. , 1998, Development.

[30]  M. Kennedy,et al.  A common precursor for hematopoietic and endothelial cells. , 1998, Development.

[31]  S. Orkin,et al.  A "knockdown" mutation created by cis-element gene targeting reveals the dependence of erythroid cell maturation on the level of transcription factor GATA-1. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[32]  J. D. Engel,et al.  GATA-1 transcription is controlled by distinct regulatory mechanisms during primitive and definitive erythropoiesis. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[33]  D. Durocher,et al.  The atrial natriuretic factor promoter is a downstream target for Nkx-2.5 in the myocardium , 1996, Molecular and cellular biology.

[34]  S. Nishikawa,et al.  In vitro analysis of epiblast tissue potency for hematopoietic cell differentiation. , 1996, Development.

[35]  A. Lassar,et al.  Induction of avian cardiac myogenesis by anterior endoderm. , 1995, Development.

[36]  Ruili Li,et al.  Myogenic and morphogenetic defects in the heart tubes of murine embryos lacking the homeo box gene Nkx2-5. , 1995, Genes & development.

[37]  Daniel J Garry,et al.  Persistent expression of the ATP-binding cassette transporter, Abcg2, identifies cardiac SP cells in the developing and adult heart. , 2004, Developmental biology.

[38]  E. Stanley,et al.  Efficient Cre-mediated deletion in cardiac progenitor cells conferred by a 3'UTR-ires-Cre allele of the homeobox gene Nkx2-5. , 2002, The International journal of developmental biology.