Endothelial CSN5 impairs NF-κB activation and monocyte adhesion to endothelial cells and is highly expressed in human atherosclerotic lesions

Yaw Asare; Erdenechimeg Shagdarsuren; Johannes A. Schmid; Pathricia V. Tilstam; Jochen Grommes; Omar El Bounkari; Anke K. Schütz; Christian Weber; Menno P. J. de Winther; Heidi Noels; Jürgen Bernhagen

doi:10.1160/TH13-02-0155

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2013; 110(01): 141-152
DOI: 10.1160/TH13-02-0155

Cardiovascular Biology and Cell Signalling

Schattauer GmbH

Endothelial CSN5 impairs NF-κB activation and monocyte adhesion to endothelial cells and is highly expressed in human atherosclerotic lesions

Authors

Yaw Asare

¹Institute of Biochemistry and Molecular Cell Biology, RWTH Aachen University, Aachen, Germany
Erdenechimeg Shagdarsuren

²Institute of Molecular Cardiovascular Research (IMCAR), RWTH Aachen University, Aachen, Germany
Johannes A. Schmid

³Center for Physiology and Pharmacology, Department of Vascular Biology and Thrombosis Research, Medical University Vienna, Vienna, Austria
Pathricia V. Tilstam

²Institute of Molecular Cardiovascular Research (IMCAR), RWTH Aachen University, Aachen, Germany
Jochen Grommes

²Institute of Molecular Cardiovascular Research (IMCAR), RWTH Aachen University, Aachen, Germany

⁴European Vascular Center Aachen-Maastricht, Department of Vascular Surgery, RWTH Aachen University, Aachen, Germany
Omar El Bounkari

¹Institute of Biochemistry and Molecular Cell Biology, RWTH Aachen University, Aachen, Germany
Anke K. Schütz

¹Institute of Biochemistry and Molecular Cell Biology, RWTH Aachen University, Aachen, Germany
Christian Weber

⁵Institute for Cardiovascular Prevention, Ludwig-Maximilians-University Munich, Munich, Germany

⁶Munich Heart Alliance, Munich, Germany

⁷Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands
Menno P. J. de Winther^*

⁷Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands
Heidi Noels^§

²Institute of Molecular Cardiovascular Research (IMCAR), RWTH Aachen University, Aachen, Germany
Jürgen Bernhagen^§

¹Institute of Biochemistry and Molecular Cell Biology, RWTH Aachen University, Aachen, Germany

⁸August-Lenz-Stiftung, Institute for Cardiovascular Research, Ludwig-Maximilians-University Munich, Munich, Germany

Abstract

Summary

The COP9 signalosome (CSN), a multifunctional protein complex involved in the regulation of cullin-RING-E3 ubiquitin ligases (CRLs), has emerged as a regulator of NF-κB signalling. As NF-κB drives the expression of pro-inflammatory and pro-atherosclerotic genes, we probed the yet unknown role of the CSN, in particular CSN5, on NF-KB-mediated atherogenic responses in endothelial cells. Co-immunoprecipitation in human umbilical vein endothelial cells (HUVECs) revealed the presence of a super-complex between IKK and CSN, which dissociates upon TNF-α stimulation. Furthermore, CSN5 silencing enhanced TNF-α-induced IKB-α degradation and NF-κB activity in luci-ferase reporter assays. This was paralleled by an increased NF-KB-driven upregulation of atherogenic chemokines and adhesion molecules, as measured by qPCR and flow cytometry, and translated into an enhanced arrest of THP-1 monocytes on TNF-α-stimulated, CSN5-depleted HUVECs. Reverse effects on NF-κB activity and THP-1 arrest were seen upon CSN5 overexpression. Finally, double-immunostaining confirmed the expression of CSN subunits in the endothelium of human atherosclerotic lesions, and revealed an increased expression of CSN5 which correlated with atheroprogression. In conclusion, endothelial CSN5 attenuates NF-KB-dependent pro-inflammatory gene expression and monocyte arrest on stimulated endothelial cells in vitro, suggesting that CSN5 might serve as a negative regulator of atherogenesis.

Note: The review process for this manuscript was fully handled by G. Y. H. Lip, Editor in Chief.

Keywords

Signal transduction - endothelial cells - cytokines - chemokines - atherosclerosis

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Referenzen

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