FoxO1 and FoxM1 Transcription Factors Have Antagonistic Functions in Neonatal Cardiomyocyte Cell-Cycle Withdrawal and IGF1 Gene Regulation

Rationale: In the mammalian heart, cardiomyocytes withdraw from the cell cycle and initiate hypertrophic growth soon after birth, but the transcriptional regulatory mechanisms that control these neonatal transitions are not well-defined. Objective: Forkhead family transcription factors have been implicated as positive (forkhead box [Fox] transcription factor M1) and negative (FoxO1 and FoxO3) regulators of cardiomyocyte proliferation prenatally, but their regulatory interactions and functions in neonatal cell-cycle withdrawal have not been reported previously. Potential regulators of Fox activity, including the metabolic indicator AMP-activated protein kinase (AMPK), and Fox transcriptional targets (p21, p27, insulin-like growth factor 1 [IGF1]) also were examined. Methods and Results: In cultured neonatal rat cardiomyocytes, AMPK activates FoxOs, and AMPK inhibition is sufficient to induce cell proliferation. In vivo, combined loss of FoxO1 and FoxO3 specifically in cardiomyocytes leads to delayed cell-cycle withdrawal and increased expression of IGF1 and FoxM1. Conversely, cardiomyocyte-specific loss of FoxM1 results in decreased neonatal cardiomyocyte cell proliferation, decreased expression of IGF1, and increased expression of cell-cycle inhibitors p21 and p27. IGF1 is a direct downstream target of cardiac Fox transcription factors, which is negatively regulated by FoxOs and positively regulated by FoxM1, dependent on AMPK activation status. Conclusions: These data support a regulatory mechanism whereby the balance of FoxO and FoxM1 transcription factors integrates metabolic status, mediated by AMPK, and cell-cycle regulation, through competitive regulation of target genes, including IGF1, in neonatal cardiomyocytes.

[1]  M. Vinciguerra,et al.  Growth Factors, Nutrient Signaling, and Cardiovascular Aging , 2012, Circulation research.

[2]  Nan Jiang,et al.  Metabolic stress-induced activation of FoxO1 triggers diabetic cardiomyopathy in mice. , 2012, The Journal of clinical investigation.

[3]  E. Lam,et al.  FOXO3a represses VEGF expression through FOXM1-dependent and -independent mechanisms in breast cancer , 2011, Oncogene.

[4]  John McAnally,et al.  Regulation of Insulin-Like Growth Factor Signaling by Yap Governs Cardiomyocyte Proliferation and Embryonic Heart Size , 2011, Science Signaling.

[5]  D. Hardie.  AMP-activated protein kinase: an energy sensor that regulates all aspects of cell function. , 2011, Genes & development.

[6]  U. Schüller,et al.  Expression of FoxM1 Is Required for the Proliferation of Medulloblastoma Cells and Indicates Worse Survival of Patients , 2011, Clinical Cancer Research.

[7]  E. Lam,et al.  FOXO and FOXM1 in cancer: the FOXO-FOXM1 axis shapes the outcome of cancer chemotherapy. , 2011, Current drug targets.

[8]  T. Kalin,et al.  Expression of Foxm1 Transcription Factor in Cardiomyocytes Is Required for Myocardial Development , 2011, PloS one.

[9]  E. Olson,et al.  Transient Regenerative Potential of the Neonatal Mouse Heart , 2011, Science.

[10]  T. Kalin,et al.  Multiple faces of FoxM1 transcription factor , 2011, Cell cycle.

[11]  M. Giacca,et al.  Knockdown of Cyclin-dependent Kinase Inhibitors Induces Cardiomyocyte Re-entry in the Cell Cycle* , 2011, The Journal of Biological Chemistry.

[12]  R. DePinho,et al.  FoxO Transcription Factors Promote Cardiomyocyte Survival upon Induction of Oxidative Stress* , 2010, The Journal of Biological Chemistry.

[13]  T. Kalin,et al.  Forkhead Box M1 Transcription Factor Is Required for Macrophage Recruitment during Liver Repair , 2010, Molecular and Cellular Biology.

[14]  Shanru Li,et al.  Foxp1 coordinates cardiomyocyte proliferation through both cell-autonomous and nonautonomous mechanisms. , 2010, Genes & development.

[15]  Elizabeth M. Mandel,et al.  The BMP pathway acts to directly regulate Tbx20 in the developing heart , 2010, Development.

[16]  James D. Johnson,et al.  AMP-activated protein kinase confers protection against TNF-{alpha}-induced cardiac cell death. , 2009, Cardiovascular research.

[17]  Arunima Sengupta,et al.  FoxO Transcription Factors Promote Autophagy in Cardiomyocytes* , 2009, The Journal of Biological Chemistry.

[18]  Yajing Wang,et al.  AMP-Activated Protein Kinase Deficiency Enhances Myocardial Ischemia/Reperfusion Injury but Has Minimal Effect on the Antioxidant/Antinitrative Protection of Adiponectin , 2009, Circulation.

[19]  J. Whitsett,et al.  Forkhead Box m1 transcription factor is required for perinatal lung function , 2008, Proceedings of the National Academy of Sciences.

[20]  U. Laufs,et al.  Metabolic Switch and Hypertrophy of Cardiomyocytes following Treatment with Angiotensin II Are Prevented by AMP-activated Protein Kinase* , 2008, Journal of Biological Chemistry.

[21]  Raymond Sawaya,et al.  FoxM1B transcriptionally regulates vascular endothelial growth factor expression and promotes the angiogenesis and growth of glioma cells. , 2008, Cancer research.

[22]  Bruce J Aronow,et al.  Shared gene expression profiles in developing heart valves and osteoblast progenitor cells. , 2008, Physiological genomics.

[23]  M. V. van Amerongen,et al.  Features of cardiomyocyte proliferation and its potential for cardiac regeneration , 2008, Journal of cellular and molecular medicine.

[24]  E. Lam,et al.  Many forks in the path: cycling with FoxO , 2008, Oncogene.

[25]  A. Brunet,et al.  The FoxO code , 2008, Oncogene.

[26]  K. Yutzey,et al.  Regulation of Cardiomyocyte Proliferation and Myocardial Growth During Development by FOXO Transcription Factors , 2008, Circulation research.

[27]  T. Mak,et al.  FOXO3a is activated in response to hypoxic stress and inhibits HIF1-induced apoptosis via regulation of CITED2. , 2007, Molecular cell.

[28]  S. Gygi,et al.  The Energy Sensor AMP-activated Protein Kinase Directly Regulates the Mammalian FOXO3 Transcription Factor* , 2007, Journal of Biological Chemistry.

[29]  F. Conlon,et al.  Cardiac Progenitors and the Embryonic Cell Cycle , 2007, Cell cycle.

[30]  M. Surani,et al.  Cdkn1c (p57Kip2) is the major regulator of embryonic growth within its imprinted domain on mouse distal chromosome 7 , 2007, BMC Developmental Biology.

[31]  K. Yutzey,et al.  Developmental regulation of the mouse IGF-I exon 1 promoter region by calcineurin activation of NFAT in skeletal muscle. , 2007, American journal of physiology. Cell physiology.

[32]  A. Malik,et al.  Myocardium defects and ventricular hypoplasia in mice homozygous null for the Forkhead Box m1 transcription factor , 2007, Developmental dynamics : an official publication of the American Association of Anatomists.

[33]  W. R. MacLellan,et al.  Cardiac myocyte cell cycle control in development, disease, and regeneration. , 2007, Physiological reviews.

[34]  Katherine E Yutzey,et al.  Tbx20 regulation of endocardial cushion cell proliferation and extracellular matrix gene expression. , 2007, Developmental biology.

[35]  Yonghong Xiao,et al.  FoxOs Are Lineage-Restricted Redundant Tumor Suppressors and Regulate Endothelial Cell Homeostasis , 2007, Cell.

[36]  D. Castrillon,et al.  Foxo Transcription Factors Blunt Cardiac Hypertrophy by Inhibiting Calcineurin Signaling , 2006, Circulation.

[37]  E. Lam,et al.  The Forkhead Box M1 Protein Regulates the Transcription of the Estrogen Receptor α in Breast Cancer Cells* , 2006, Journal of Biological Chemistry.

[38]  E. Araki,et al.  AMPK and cell proliferation – AMPK as a therapeutic target for atherosclerosis and cancer , 2006, The Journal of physiology.

[39]  R. Schwartz,et al.  Cardiac-Specific Deletion of Gata4 Reveals Its Requirement for Hypertrophy, Compensation, and Myocyte Viability , 2006, Circulation research.

[40]  T. Asano,et al.  Adenosine Monophosphate-Activated Protein Kinase Suppresses Vascular Smooth Muscle Cell Proliferation Through the Inhibition of Cell Cycle Progression , 2005, Circulation research.

[41]  A. Goldberg,et al.  The FOXO3a Transcription Factor Regulates Cardiac Myocyte Size Downstream of AKT Signaling* , 2005, Journal of Biological Chemistry.

[42]  T. Unterman,et al.  FoxO proteins in insulin action and metabolism , 2005, Trends in Endocrinology & Metabolism.

[43]  K. Kaestner,et al.  The mouse Forkhead Box m1 transcription factor is essential for hepatoblast mitosis and development of intrahepatic bile ducts and vessels during liver morphogenesis. , 2004, Developmental biology.

[44]  J. Neilson,et al.  Genetic Loss of Calcineurin Blocks Mechanical Overload-induced Skeletal Muscle Fiber Type Switching but Not Hypertrophy* , 2004, Journal of Biological Chemistry.

[45]  W. Aird,et al.  The Akt-regulated Forkhead Transcription Factor FOXO3a Controls Endothelial Cell Viability through Modulation of the Caspase-8 Inhibitor FLIP* , 2004, Journal of Biological Chemistry.

[46]  M. Matsui,et al.  In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. , 2003, Molecular biology of the cell.

[47]  L. Field,et al.  Cardiomyocyte cell cycle regulation. , 2002, Circulation research.

[48]  D. Accili,et al.  The forkhead transcription factor Foxo1 (Fkhr) confers insulin sensitivity onto glucose-6-phosphatase expression. , 2001, The Journal of clinical investigation.

[49]  J. Molkentin,et al.  Calcineurin Promotes Protein Kinase C and c-Jun NH2-terminal Kinase Activation in the Heart , 2000, The Journal of Biological Chemistry.

[50]  Johannes Gerdes,et al.  The Ki‐67 protein: From the known and the unknown , 2000, Journal of cellular physiology.

[51]  P. Anversa,et al.  Insulin-like growth factor-1 receptor and its ligand regulate the reentry of adult ventricular myocytes into the cell cycle. , 1997, Experimental cell research.

[52]  H. Clevers,et al.  The winged-helix transcription factor Trident is expressed in cycling cells. , 1997, Nucleic acids research.

[53]  A. Gerdes,et al.  Rapid transition of cardiac myocytes from hyperplasia to hypertrophy during postnatal development. , 1996, Journal of molecular and cellular cardiology.