Adenosine-to-inosine RNA editing controls cathepsin S expression in atherosclerosis by enabling HuR-mediated post-transcriptional regulation.
Adenosine-to-inosine (A-to-I) RNA editing, which is catalyzed by a family of adenosine deaminase acting on RNA (ADAR) enzymes, is important in the epitranscriptomic regulation of RNA metabolism. However, the role of A-to-I RNA editing in vascular disease is unknown. Here we show that cathepsin S mRNA (CTSS), which encodes a cysteine protease associated with angiogenesis and atherosclerosis, is highly edited in human endothelial cells. The 3' untranslated region (3' UTR) of the CTSS transcript contains two inverted repeats, the AluJo and AluSx+regions, which form a long stem-loop structure that is recognized by ADAR1 as a substrate for editing. RNA editing enables the recruitment of the stabilizing RNA-binding protein human antigen R (HuR; encoded by ELAVL1) to the 3' UTR of the CTSS transcript, thereby controlling CTSS mRNA stability and expression. In endothelial cells, ADAR1 overexpression or treatment of cells with hypoxia or with the inflammatory cytokines interferon-γ and tumor-necrosis-factor-α induces CTSS RNA editing and consequently increases cathepsin S expression. ADAR1 levels and the extent of CTSS RNA editing are associated with changes in cathepsin S levels in patients with atherosclerotic vascular diseases, including subclinical atherosclerosis, coronary artery disease, aortic aneurysms and advanced carotid atherosclerotic disease. These results reveal a previously unrecognized role of RNA editing in gene expression in human atherosclerotic vascular diseases.
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|Department of Cardiology, Center of Internal Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Konstantinos
K
Stellos
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Aikaterini
A
Gatsiou
Department of Clinical Therapeutics, Alexandra Hospital, National and Kapodistrian University of Athens, Athens, Greece.
Kimon
K
Stamatelopoulos
Department of Molecular Medicine and Surgery, Karolinska Institute, Stockholm, Sweden.
Ljubica
L
Perisic Matic
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
David
D
John
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Federica Francesca
FF
Lunella
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Nicolas
N
Jaé
Institute of Biochemistry, University of Giessen, Giessen, Germany.
Oliver
O
Rossbach
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.
Carolin
C
Amrhein
Department of Vascular Surgery, 1st Propaedeutic Department of Surgery, Hippocratio General Hospital, National and Kapodistrian University of Athens, Athens, Greece.
Frangiska
F
Sigala
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Reinier A
0000-0001-7944-9748
RA
Boon
Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance, Goethe University Frankfurt, Frankfurt, Germany.
Boris
B
Fürtig
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Yosif
Y
Manavski
Laboratory of Functional Genomics and Systems Biology, Max Delbrück Center for Molecular Medicine Berlin-Buch, Berlin, Germany.
Xintian
X
You
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Shizuka
S
Uchida
Department of Cardiology, Center of Internal Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Till
T
Keller
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Jes-Niels
JN
Boeckel
Cardiothoracic Surgery Unit, Department of Molecular Medicine and Surgery, Karolinska Institute, Stockholm, Sweden.
Anders
A
Franco-Cereceda
Cardiovascular Medicine Unit, Department of Medicine, Karolinska Institute, Stockholm, Sweden.|Department of Vascular and Endovascular Surgery, Technical University Munich, Munich, Germany.
Lars
0000-0001-5228-2634
L
Maegdefessel
Laboratory of Functional Genomics and Systems Biology, Max Delbrück Center for Molecular Medicine Berlin-Buch, Berlin, Germany.
Wei
W
Chen
Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance, Goethe University Frankfurt, Frankfurt, Germany.
Harald
H
Schwalbe
Institute of Biochemistry, University of Giessen, Giessen, Germany.
Albrecht
A
Bindereif
Cardiovascular Medicine Unit, Department of Medicine, Karolinska Institute, Stockholm, Sweden.
Per
P
Eriksson
Department of Molecular Medicine and Surgery, Karolinska Institute, Stockholm, Sweden.
Ulf
0000-0001-9212-3945
U
Hedin
Department of Cardiology, Center of Internal Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Andreas M
AM
Zeiher
Institute of Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany.|German Center of Cardiovascular Research (Deutsches Zentrum für Herz-Kreislaufforschung; DZHK), Rhein-Main Partner Site, Frankfurt, Germany.
Stefanie
S
Dimmeler
D020413:3' Untranslated Regions; D000241:Adenosine / Q000378:metabolism; D000243:Adenosine Deaminase / Q000235:genetics*; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001014:Aortic Aneurysm / Q000235:genetics; D050197:Atherosclerosis / Q000235:genetics*; D002340:Carotid Artery Diseases / Q000235:genetics; D002403:Cathepsins / Q000235:genetics*; D003324:Coronary Artery Disease / Q000235:genetics; D000067780:ELAV-Like Protein 1 / Q000235:genetics*; D005260:Female; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D055879:Gene Knock-In Techniques; D055785:Gene Knockdown Techniques; D059014:High-Throughput Nucleotide Sequencing; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D000860:Hypoxia / Q000235:genetics; D015151:Immunoblotting; D007288:Inosine / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D017393:RNA Editing / Q000187:drug effects / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D020413:3' Untranslated Regions; D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D014409:Tumor Necrosis Factor-alpha; D007288:Inosine; D007371:Interferon-gamma; D002403:Cathepsins; C028217:cathepsin S; C000595356:ADAR protein, human; D000243:Adenosine Deaminase; D000241:Adenosine