title: C2238/αANP modulates apolipoprotein E through Egr-1/miR199a in vascular smooth muscle cells in vitro.
abstract: Subjects carrying the T2238C ANP gene variant have a higher risk to suffer a stroke or myocardial infarction. The mechanisms through which T2238C/αANP exerts detrimental vascular effects need to be fully clarified. In the present work we aimed at exploring the impact of C2238/αANP (mutant form) on atherosclerosis-related pathways. As a first step, an atherosclerosis gene expression macroarray analysis was performed in vascular smooth muscle cells (VSMCs) exposed to either T2238/αANP (wild type) or C2238/αANP. The major finding was that apolipoprotein E (ApoE) gene expression was significantly downregulated by C2238/αANP and it was upregulated by T2238/αANP. We subsequently found that C2238/αANP induces ApoE downregulation through type C natriuretic peptide receptor (NPR-C)-dependent mechanisms involving the upregulation of miR199a-3p and miR199a-5p and the downregulation of DNAJA4. In fact, NPR-C knockdown rescued ApoE level. Upregulation of miR199a by NPR-C was mediated by a reactive oxygen species-dependent increase of the early growth response protein-1 (Egr-1) transcription factor. In fact, Egr-1 knockdown abolished the impact of C2238/αANP on ApoE and miR199a. Of note, downregulation of ApoE by C2238/αANP was associated with a significant increase in inflammation, apoptosis and necrosis that was completely rescued by the exogenous administration of recombinant ApoE. In conclusion, our study dissected a novel mechanism of vascular damage exerted by C2238/αANP that is mediated by ApoE downregulation. We provide the first demonstration that C2238/αANP downregulates ApoE in VSMCs through NPR-C-dependent activation of Egr-1 and the consequent upregulation of miR199a. Restoring ApoE levels could represent a potential therapeutic strategy to counteract the harmful effects of C2238/αANP.
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Stanzione
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.|Department of Medical-Surgical Sciences and Biotechnologies, Sapienza University of Rome, Latina, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Sciarretta
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Marchitti
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Bianchi
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Di Castro
authors.affiliation: Department of Clinical and Molecular Medicine, School of Medicine and Psychology, Sapienza University of Rome, Ospedale S. Andrea, Rome, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Scarpino
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Cotugno
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.|Department of Medical-Surgical Sciences and Biotechnologies, Sapienza University of Rome, Latina, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Frati
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.|Department of Clinical and Molecular Medicine, School of Medicine and Psychology, Sapienza University of Rome, Ospedale S. Andrea, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Volpe
authors.affiliation: IRCCS Neuromed, Pozzilli (Is), Sapienza University of Rome, Latina, Italy.|Department of Clinical and Molecular Medicine, School of Medicine and Psychology, Sapienza University of Rome, Ospedale S. Andrea, Rome, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Rubattu
mesh_terms: D001057:Apolipoproteins E / Q000235:genetics / Q000378:metabolism*; D017209:Apoptosis; D050197:Atherosclerosis / Q000235:genetics / Q000473:pathology; D009320:Atrial Natriuretic Factor / Q000235:genetics*; D015415:Biomarkers / Q000378:metabolism; D002470:Cell Survival; D003331:Coronary Vessels / Q000166:cytology; D051766:Early Growth Response Protein 1 / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000473:pathology; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D009154:Mutation / Q000235:genetics*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D009336:Necrosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D017461:Receptors, Atrial Natriuretic Factor / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology; D015854:Up-Regulation
chemical_list: D001057:Apolipoproteins E; D015415:Biomarkers; D051766:Early Growth Response Protein 1; D035683:MicroRNAs; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; C531408:mirn199 microRNA, human; D009320:Atrial Natriuretic Factor; D017461:Receptors, Atrial Natriuretic Factor