The C2238/αANP variant is a negative modulator of both viability and function of coronary artery smooth muscle cells.
BACKGROUND
Abnormalities of vascular smooth muscle cells (VSMCs) contribute to development of vascular disease. Atrial natriuretic peptide (ANP) exerts important effects on VSMCs. A common ANP molecular variant (T2238C/αANP) has recently emerged as a novel vascular risk factor.


OBJECTIVE
We aimed at identifying effects of CC2238/αANP on viability, migration and motility in coronary artery SMCs, and the underlying signaling pathways.


RESULTS
Cells were exposed to either TT2238/αANP or CC2238/αANP. At the end of treatment, cell viability, migration and motility were evaluated, along with changes in oxidative stress pathway (ROS levels, NADPH and eNOS expression), on Akt phosphorylation and miR21 expression levels. CC2238/αANP reduced cell vitality, increased apoptosis and necrosis, increased oxidative stress levels, suppressed miR21 expression along with consistent changes of its molecular targets (PDCD4, PTEN, Bcl2) and of phosphorylated Akt levels. As a result of increased oxidative stress, CC2238/αANP markedly stimulated cell migration and increased cell contraction. NPR-C gene silencing with specific siRNAs restored cell viability, miR21 expression, and reduced oxidative stress induced by CC2238/αANP. The cAMP/PKA/CREB pathway, driven by NPR-C activation, significantly contributed to both miR21 and phosphoAkt reduction upon CC2238/αANP. miR21 overexpression by mimic-hsa-miR21 rescued the cellular damage dependent on CC2238/αANP.


CONCLUSIONS
CC2238/αANP negatively modulates viability through NPR-C/cAMP/PKA/CREB/miR21 signaling pathway, and it augments oxidative stress leading to increased migratory and vasoconstrictor effects in coronary artery SMCs. These novel findings further support a damaging role of this common αANP variant on vessel wall and its potential contribution to acute coronary events.
IRCCS Neuromed, Pozzilli (Isernia), Italy; Department of Clinical and Molecular Medicine, School of Medicine and Psychology, University Sapienza of Rome, Ospedale S. Andrea, Rome, Italy.
Speranza
S
Rubattu
IRCCS Neuromed, Pozzilli (Isernia), Italy.
Simona
S
Marchitti
IRCCS Neuromed, Pozzilli (Isernia), Italy.
Franca
F
Bianchi
IRCCS Neuromed, Pozzilli (Isernia), Italy.
Sara
S
Di Castro
IRCCS Neuromed, Pozzilli (Isernia), Italy.
Rosita
R
Stanzione
IRCCS Neuromed, Pozzilli (Isernia), Italy.
Maria
M
Cotugno
Department of Clinical and Molecular Medicine, School of Medicine and Psychology, University Sapienza of Rome, Ospedale S. Andrea, Rome, Italy.
Cristina
C
Bozzao
IRCCS Neuromed, Pozzilli (Isernia), Italy.
Sebastiano
S
Sciarretta
IRCCS Neuromed, Pozzilli (Isernia), Italy; Department of Clinical and Molecular Medicine, School of Medicine and Psychology, University Sapienza of Rome, Ospedale S. Andrea, Rome, Italy.
Massimo
M
Volpe
D017209:Apoptosis / Q000187:drug effects*; D009320:Atrial Natriuretic Factor / Q000235:genetics / Q000494:pharmacology*; D015153:Blotting, Western; D002097:C-Reactive Protein / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D000242:Cyclic AMP / Q000494:pharmacology; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D035683:MicroRNAs / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D011110:Polymorphism, Genetic / Q000235:genetics*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects*; D000099153:Pentraxins
D009320:Atrial Natriuretic Factor; D002097:C-Reactive Protein; D000242:Cyclic AMP; D017362:Cyclic AMP Response Element-Binding Protein; D017868:Cyclic AMP-Dependent Protein Kinases; D035683:MicroRNAs; D009419:Nerve Tissue Proteins; D051057:Proto-Oncogene Proteins c-akt; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D000099153:Pentraxins; C115163:CREB1 protein, human; C524940:MIRN21 microRNA, human