title: Oxidative Stress-Induced miR-200c Disrupts the Regulatory Loop Among SIRT1, FOXO1, and eNOS.
abstract: Reactive oxygen species (ROS) play a pivotal role in different pathologic conditions, including ischemia, diabetes, and aging. We previously showed that ROS enhance miR-200c expression, causing endothelial cell (EC) apoptosis and senescence. Herein, we dissect the interaction among miR-200c and three strictly related proteins that modulate EC function and ROS production: sirtuin 1 (SIRT1), endothelial nitric oxide synthase (eNOS), and forkhead box O1 (FOXO1). Moreover, the role of miR-200c on ROS modulation was also investigated.
  
  
  
  We demonstrated that miR-200c directly targets SIRT1, eNOS, and FOXO1; via this mechanism, miR-200c decreased NO and increased the acetylation of SIRT1 targets, that is, FOXO1 and p53. FOXO1 acetylation inhibited its transcriptional activity on target genes, that is, SIRT1 and the ROS scavengers, catalase and manganese superoxide dismutase. In keeping, miR-200c increased ROS production and induced p66Shc protein phosphorylation in Ser-36; this mechanism upregulated ROS and inhibited FOXO1 transcription, reinforcing this molecular circuitry. These in vitro results were validated in three in vivo models of oxidative stress, that is, human skin fibroblasts from old donors, femoral arteries from old mice, and a murine model of hindlimb ischemia. In all cases, miR-200c was higher versus control and its targets, that is, SIRT1, eNOS, and FOXO1, were downmodulated. In the mouse hindlimb ischemia model, anti-miR-200c treatment rescued these targets and improved limb perfusion. Innovation and Conclusion: miR-200c disrupts SIRT1/FOXO1/eNOS regulatory loop. This event promotes ROS production and decreases NO, contributing to endothelial dysfunction under conditions of increased oxidative stress such as aging and ischemia. Antioxid. Redox Signal. 27, 328-344.
authors.affiliation: 1 Vascular Pathology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Carlomosti
authors.affiliation: 2 Department of Experimental Medicine, University of Rome , Sapienza, Rome, Italy .
authors.forename: Marco
authors.initials: M
authors.lastname: D'Agostino
authors.affiliation: 1 Vascular Pathology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Sara
authors.initials: S
authors.lastname: Beji
authors.affiliation: 3 Department of Biology and Biotechnology "Charles Darwin," Sapienza University , Rome, Italy .|4 Institute of Cell Biology and Neurobiology (IBCN) , National Research Council of Italy (CNR), Rome, Italy .
authors.forename: Alessio
authors.initials: A
authors.lastname: Torcinaro
authors.affiliation: 4 Institute of Cell Biology and Neurobiology (IBCN) , National Research Council of Italy (CNR), Rome, Italy .
authors.forename: Roberto
authors.initials: R
authors.lastname: Rizzi
authors.affiliation: 5 Molecular Cardiology Laboratory, IRCCS-Policlinico San Donato , Milan, Italy .
authors.forename: Germana
authors.initials: G
authors.lastname: Zaccagnini
authors.affiliation: 5 Molecular Cardiology Laboratory, IRCCS-Policlinico San Donato , Milan, Italy .
authors.forename: Biagina
authors.initials: B
authors.lastname: Maimone
authors.affiliation: 1 Vascular Pathology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Valeria
authors.initials: V
authors.lastname: Di Stefano
authors.affiliation: 4 Institute of Cell Biology and Neurobiology (IBCN) , National Research Council of Italy (CNR), Rome, Italy .|6 Santa Lucia Foundation-FSL-IRCCS , Rome, Italy .
authors.forename: Francesca
authors.initials: F
authors.lastname: De Santa
authors.affiliation: 7 Molecular and Cell Biology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Sonia
authors.initials: S
authors.lastname: Cordisco
authors.affiliation: 1 Vascular Pathology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Annalisa
authors.initials: A
authors.lastname: Antonini
authors.affiliation: 1 Vascular Pathology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Roberta
authors.initials: R
authors.lastname: Ciarapica
authors.affiliation: 7 Molecular and Cell Biology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Elena
authors.initials: E
authors.lastname: Dellambra
authors.affiliation: 5 Molecular Cardiology Laboratory, IRCCS-Policlinico San Donato , Milan, Italy .
authors.forename: Fabio
authors.initials: F
authors.lastname: Martelli
authors.affiliation: 8 Unità di Biologia Vascolare e Medicina Rigenerativa, Centro Cardiologico Monzino, IRCCS , Milan, Italy .
authors.forename: Daniele
authors.initials: D
authors.lastname: Avitabile
authors.affiliation: 1 Vascular Pathology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Maurizio Colognesi
authors.initials: MC
authors.lastname: Capogrossi
authors.affiliation: 1 Vascular Pathology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS , FLMM, Rome, Italy .
authors.forename: Alessandra
authors.initials: A
authors.lastname: Magenta
mesh_terms: D000107:Acetylation; D000818:Animals; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D000071161:Forkhead Box Protein O1 / Q000378:metabolism*; D057809:HEK293 Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D009569:Nitric Oxide / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D018384:Oxidative Stress; D010766:Phosphorylation; D017382:Reactive Oxygen Species / Q000378:metabolism; D056564:Sirtuin 1 / Q000235:genetics* / Q000378:metabolism; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1 / Q000378:metabolism
chemical_list: C085126:FOXO1 protein, human; D000071161:Forkhead Box Protein O1; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D017382:Reactive Oxygen Species; C528024:SHC1 protein, human; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D009569:Nitric Oxide; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; C447939:SIRT1 protein, human; D056564:Sirtuin 1
keywords: aging; free radicals; microRNA; nitric oxide; vascular