title: Ezh2-mediated repression of a transcriptional pathway upstream of Mmp9 maintains integrity of the developing vasculature.
abstract: Maintenance of vascular integrity is required for embryogenesis and organ homeostasis. However, the gene expression programs that stabilize blood vessels are poorly understood. Here, we show that the histone methyltransferase Ezh2 maintains integrity of the developing vasculature by repressing a transcriptional program that activates expression of Mmp9. Inactivation of Ezh2 in developing mouse endothelium caused embryonic lethality with compromised vascular integrity and increased extracellular matrix degradation. Genome-wide approaches showed that Ezh2 targets Mmp9 and its activators Fosl1 and Klf5. In addition, we uncovered Creb3l1 as an Ezh2 target that directly activates Mmp9 gene expression in the endothelium. Furthermore, genetic inactivation of Mmp9 rescued vascular integrity defects in Ezh2-deficient embryos. Thus, epigenetic repression of Creb3l1, Fosl1, Klf5 and Mmp9 by Ezh2 in endothelial cells maintains the integrity of the developing vasculature, potentially linking this transcriptional network to diseases with compromised vascular integrity.
authors.affiliation: Gladstone Institute of Cardiovascular Disease, and Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, CA 94158, USA Program in Physiology and Experimental Medicine, The Hospital for Sick Children, Toronto, Ontario M5G0A4, Canada Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5S 1A8, Canada Heart and Stroke Richard Lewar Centre of Excellence, Toronto, Ontario M5G 1L7, Canada paul.delgadoolguin@sickkids.ca bbruneau@gladstone.ucsf.edu.
authors.forename: Paul
authors.initials: P
authors.lastname: Delgado-Olguín
authors.affiliation: Heart and Stroke Richard Lewar Centre of Excellence, Toronto, Ontario M5G 1L7, Canada Toronto General Research Institute, University Health Network, Toronto, Ontario M5G 1L7, Canada Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario M5S 1A1, Canada.
authors.forename: Lan T
authors.initials: LT
authors.lastname: Dang
authors.affiliation: Gladstone Institute of Cardiovascular Disease, and Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, CA 94158, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: He
authors.affiliation: Gladstone Institute of Cardiovascular Disease, and Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, CA 94158, USA.
authors.forename: Sean
authors.initials: S
authors.lastname: Thomas
authors.affiliation: Program in Physiology and Experimental Medicine, The Hospital for Sick Children, Toronto, Ontario M5G0A4, Canada.
authors.forename: Lijun
authors.initials: L
authors.lastname: Chi
authors.affiliation: Gladstone Institute of Cardiovascular Disease, and Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, CA 94158, USA.
authors.forename: Tatyana
authors.initials: T
authors.lastname: Sukonnik
authors.affiliation: Heart and Stroke Richard Lewar Centre of Excellence, Toronto, Ontario M5G 1L7, Canada Toronto General Research Institute, University Health Network, Toronto, Ontario M5G 1L7, Canada Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario M5S 1A1, Canada.
authors.forename: Nadiya
authors.initials: N
authors.lastname: Khyzha
authors.affiliation: Laboratory of Lymphocyte Signaling, The Rockefeller University, New York, NY 10021, USA.
authors.forename: Marc-Werner
authors.initials: MW
authors.lastname: Dobenecker
authors.affiliation: Heart and Stroke Richard Lewar Centre of Excellence, Toronto, Ontario M5G 1L7, Canada Toronto General Research Institute, University Health Network, Toronto, Ontario M5G 1L7, Canada Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario M5S 1A1, Canada.
authors.forename: Jason E
authors.initials: JE
authors.lastname: Fish
authors.affiliation: Gladstone Institute of Cardiovascular Disease, and Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, CA 94158, USA Department of Pediatrics, Cardiovascular Research Institute, and Institute for Regeneration Medicine, University of California San Francisco, San Francisco, CA 94158, USA paul.delgadoolguin@sickkids.ca bbruneau@gladstone.ucsf.edu.
authors.forename: Benoit G
authors.initials: BG
authors.lastname: Bruneau
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D001808:Blood Vessels / Q000196:embryology*; D015153:Blotting, Western; D047369:Chromatin Immunoprecipitation; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D003959:Diamines; D000071221:Enhancer of Zeste Homolog 2 Protein; D063185:Epigenetic Repression / Q000235:genetics / Q000502:physiology*; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D017403:In Situ Hybridization; D051741:Kruppel-Like Transcription Factors; D008156:Luciferases; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D046529:Microscopy, Electron, Transmission; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009930:Organic Chemicals; D063151:Polycomb Repressive Complex 2 / Q000235:genetics / Q000378:metabolism*; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D015398:Signal Transduction / Q000502:physiology*; D000099169:Fos-Related Antigen 1
chemical_list: D052160:Benzothiazoles; D017362:Cyclic AMP Response Element-Binding Protein; D017931:DNA Primers; D003959:Diamines; D000071221:Enhancer of Zeste Homolog 2 Protein; D051741:Kruppel-Like Transcription Factors; D008156:Luciferases; D020780:Matrix Metalloproteinase 9; D009419:Nerve Tissue Proteins; D009930:Organic Chemicals; D063151:Polycomb Repressive Complex 2; D016760:Proto-Oncogene Proteins c-fos; D011804:Quinolines; D000099169:Fos-Related Antigen 1; C119923:Creb3l1 protein, mouse; C118391:Klf5 protein, mouse; C098022:SYBR Green I; C498410:Ezh2 protein, mouse; C501830:Mmp9 protein, mouse
keywords: Endothelium; Epigenetics; Extracellular matrix; Ezh2; Histone methylation; Mmp9; Mouse; Vascular development; Vascular stability