title: EZH2 modulates angiogenesis in vitro and in a mouse model of limb ischemia.
abstract: Epigenetic mechanisms may regulate the expression of pro-angiogenic genes, thus affecting reparative angiogenesis in ischemic limbs. The enhancer of zest homolog-2 (EZH2) induces thtrimethylation of lysine 27 on histone H3 (H3K27me3), which represses gene transcription. We explored (i) if EZH2 expression is regulated by hypoxia and ischemia; (ii) the impact of EZH2 on the expression of two pro-angiogenic genes: eNOS and BDNF; (iii) the functional effect of EZH2 inhibition on cultured endothelial cells (ECs); (iv) the therapeutic potential of EZH2 inhibition in a mouse model of limb ischemia (LI). EZH2 expression was increased in cultured ECs exposed to hypoxia (control: normoxia) and in ECs extracted from mouse ischemic limb muscles (control: absence of ischemia). EZH2 increased the H3K27me3 abundance onto regulatory regions of eNOS and BDNF promoters. In vitro RNA silencing or pharmacological inhibition by 3-deazaneplanocin (DZNep) of EZH2 increased eNOS and BDNF mRNA and protein levels and enhanced functional capacities (migration, angiogenesis) of ECs under either normoxia or hypoxia. In mice with experimentally induced LI, DZNep increased angiogenesis in ischaemic muscles, the circulating levels of pro-angiogenic hematopoietic cells and blood flow recovery. Targeting EZH2 for inhibition may open new therapeutic avenues for patients with limb ischemia.
authors.affiliation: Bristol Heart Institute, School of Clinical Sciences, University of Bristol, England, UK.
authors.forename: Tijana
authors.initials: T
authors.lastname: Mitić
authors.affiliation: 1] Bristol Heart Institute, School of Clinical Sciences, University of Bristol, England, UK [2] Center for Cardiovascular Sciences, Queen's Medical Research Institute, University of Edinburgh, Scotland, UK.
authors.forename: Andrea
authors.initials: A
authors.lastname: Caporali
authors.affiliation: Bristol Heart Institute, School of Clinical Sciences, University of Bristol, England, UK.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Floris
authors.affiliation: Bristol Heart Institute, School of Clinical Sciences, University of Bristol, England, UK.
authors.forename: Marco
authors.initials: M
authors.lastname: Meloni
authors.affiliation: Bristol Heart Institute, School of Clinical Sciences, University of Bristol, England, UK.
authors.forename: Micol
authors.initials: M
authors.lastname: Marchetti
authors.affiliation: Laboratory of Epigenetics and Chromatin Dynamics, Mayo Clinic, Rochester, Minnesota, USA.
authors.forename: Raul
authors.initials: R
authors.lastname: Urrutia
authors.affiliation: 1] Bristol Heart Institute, School of Clinical Sciences, University of Bristol, England, UK [2] National Heart and Lung Institute, Hammersmith Campus, Imperial College of London, London, England, UK.
authors.forename: Gianni D
authors.initials: GD
authors.lastname: Angelini
authors.affiliation: 1] Bristol Heart Institute, School of Clinical Sciences, University of Bristol, England, UK [2] National Heart and Lung Institute, Hammersmith Campus, Imperial College of London, London, England, UK.
authors.forename: Costanza
authors.initials: C
authors.lastname: Emanueli
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D015687:Cell Hypoxia; D000071221:Enhancer of Zeste Homolog 2 Protein; D044127:Epigenesis, Genetic; D005263:Femoral Artery / Q000601:surgery; D006614:Hindlimb / Q000098:blood supply / Q000187:drug effects / Q000601:surgery; D006657:Histones / Q000235:genetics / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D000860:Hypoxia / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D007511:Ischemia / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D063151:Polycomb Repressive Complex 2 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D006657:Histones; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C048460:3-deazaneplanocin; C579455:BDNF protein, human; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; C498409:EZH2 protein, human; D000071221:Enhancer of Zeste Homolog 2 Protein; D063151:Polycomb Repressive Complex 2; D000241:Adenosine