title: Epigenetic histone modification and cardiovascular lineage programming in mouse embryonic stem cells exposed to laminar shear stress.
abstract: Experimental evidence indicates that shear stress (SS) exerts a morphogenetic function during cardiac development of mouse and zebrafish embryos. However, the molecular basis for this effect is still elusive. Our previous work described that in adult endothelial cells, SS regulates gene expression by inducing epigenetic modification of histones and activation of transcription complexes bearing acetyltransferase activity. In this study, we evaluated whether SS treatment could epigenetically modify histones and influence cell differentiation in mouse embryonic stem (ES) cells. Cells were exposed to a laminar SS of 10 dyne per cm2/s(-1), or kept in static conditions in the presence or absence of the histone deacetylase inhibitor trichostatin A (TSA). These experiments revealed that SS enhanced lysine acetylation of histone H3 at position 14 (K14), as well as serine phosphorylation at position 10 (S10) and lysine methylation at position 79 (K79), and cooperated with TSA, inducing acetylation of histone H4 and phosphoacetylation of S10 and K14 of histone H3. In addition, ES cells exposed to SS strongly activated transcription from the vascular endothelial growth factor (VEGF) receptor 2 promoter. This effect was paralleled by an early induction of cardiovascular markers, including smooth muscle actin, smooth muscle protein 22-alpha, platelet-endothelial cell adhesion molecule-1, VEGF receptor 2, myocyte enhancer factor-2C (MEF2C), and alpha-sarcomeric actin. In this condition, transcription factors MEF2C and Sma/MAD homolog protein 4 could be isolated from SS-treated ES cells complexed with the cAMP response element-binding protein acetyltransferase. These results provide molecular basis for the SS-dependent cardiovascular commitment of mouse ES cells and suggest that laminar flow may be successfully applied for the in vitro production of cardiovascular precursors.
authors.affiliation: Laboratorio di Biologia Vascolare e Terapia Genica, Centro Cardiologico Fondazione I. Monzino IRCCS, Milan, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Illi
authors.forename: Alessandro
authors.initials: A
authors.lastname: Scopece
authors.forename: Simona
authors.initials: S
authors.lastname: Nanni
authors.forename: Antonella
authors.initials: A
authors.lastname: Farsetti
authors.forename: Liliana
authors.initials: L
authors.lastname: Morgante
authors.forename: Paolo
authors.initials: P
authors.lastname: Biglioli
authors.forename: Maurizio C
authors.initials: MC
authors.lastname: Capogrossi
authors.forename: Carlo
authors.initials: C
authors.lastname: Gaetano
mesh_terms: D000107:Acetylation; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage; D002478:Cells, Cultured / Q000166:cytology / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000502:physiology; D044127:Epigenesis, Genetic; D005318:Fetal Heart / Q000166:cytology*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000502:physiology; D006657:Histones / Q000502:physiology*; D006877:Hydroxamic Acids / Q000494:pharmacology; D064326:MEF2 Transcription Factors; D008745:Methylation; D051379:Mice; D009024:Morphogenesis; D009124:Muscle Proteins / Q000096:biosynthesis / Q000235:genetics; D018006:Myogenic Regulatory Factors / Q000502:physiology; D009687:Nuclear Proteins / Q000502:physiology; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D051901:Smad4 Protein; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013314:Stress, Mechanical; D015534:Trans-Activators / Q000502:physiology; D014157:Transcription Factors / Q000502:physiology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics; D048051:p38 Mitogen-Activated Protein Kinases / Q000502:physiology
chemical_list: D004268:DNA-Binding Proteins; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; D064326:MEF2 Transcription Factors; C578041:Mef2c protein, mouse; D009124:Muscle Proteins; D018006:Myogenic Regulatory Factors; D009687:Nuclear Proteins; D051901:Smad4 Protein; C490810:Smad4 protein, mouse; D015534:Trans-Activators; D014157:Transcription Factors; C012589:trichostatin A; D040301:Vascular Endothelial Growth Factor Receptor-2; D048051:p38 Mitogen-Activated Protein Kinases; D006655:Histone Deacetylases