C2238/αANP modulates apolipoprotein E through Egr-1/miR199a in vascular smooth muscle cells in vitro

Cell Death Dis. 2015 Dec 31;6(12):e2033. doi: 10.1038/cddis.2015.370.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apolipoproteins E / genetics
  • Apolipoproteins E / metabolism*
  • Apoptosis
  • Atherosclerosis / genetics
  • Atherosclerosis / pathology
  • Atrial Natriuretic Factor / genetics*
  • Biomarkers / metabolism
  • Cell Survival
  • Coronary Vessels / cytology
  • Early Growth Response Protein 1 / metabolism*
  • Humans
  • Inflammation / pathology
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Models, Biological
  • Muscle, Smooth, Vascular / cytology*
  • Mutation / genetics*
  • Myocytes, Smooth Muscle / metabolism*
  • Necrosis
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reactive Oxygen Species / metabolism
  • Real-Time Polymerase Chain Reaction
  • Receptors, Atrial Natriuretic Factor / metabolism
  • Umbilical Veins / cytology
  • Up-Regulation

Substances

  • Apolipoproteins E
  • Biomarkers
  • Early Growth Response Protein 1
  • MicroRNAs
  • RNA, Messenger
  • Reactive Oxygen Species
  • mirn199 microRNA, human
  • Atrial Natriuretic Factor
  • Receptors, Atrial Natriuretic Factor