Loss of p27 phosphorylation at Ser10 accelerates early atherogenesis by promoting leukocyte recruitment via RhoA/ROCK. (July 2015)

• Record Type: Journal Article
• Authors: Molina-Sánchez, P.; Chèvre, R.; Rius, C.; Fuster, J.J.; Andrés, V.
• Is Part Of: Journal of molecular and cellular cardiology. Volume 84(2015:Jul.)
• Page Start: 84
• Page End: 94
• Publication Date: 2015-07
• Subjects: apoE−/− apolipoprotein E-null -- apoE−/−p27Ser10Ala apolipoprotein E-null p27 Serine-10-Alanine -- BM bone marrow -- CKI cyclin-dependent kinase inhibitor -- DMEM Dulbecco's modified eagle medium -- EC(s) endothelial cell(s) -- ECGS endothelial cell growth supplement -- EDTA ethylenediaminetetraacetic acid -- ERM ezrin/radixin/moesin -- E-Sel E-selectin -- FBS fetal bovine serum -- GFP green fluorescent protein -- HBSS Hank's Balanced Salt Solution -- HFD high fat diet -- HRP horseradish peroxidase -- ICAM-1 intercellular adhesion molecule-1 -- IV intravenous -- L-Sel L-selectin -- mAECs mouse aorta endothelial cells -- Mafia Macrophage Fas-Induced Apoptosis -- P-Sel P-selectin -- p27 p27Kip1 -- p27-phospho-Ser-10 p27 phosphorylation at serine 10 -- p27Ser10Ala p27 serine-10-alanine -- PBS phosphate buffered saline -- PCR polymerase chain reaction -- qPCR quantitative real-time PCR -- ROCK Rho-associated coiled-coil containing protein kinase -- Ser10 serine 10 -- SDS sodium dodecyl sulfate -- TNFα tumor necrosis factor-α -- VCAM-1 vascular cell adhesion molecule-1; Atherosclerosis -- Endothelial cell -- Leukocyte recruitment -- p27 -- RhoA; Cardiology -- Periodicals; Heart Diseases -- Periodicals; Molecular Biology -- Periodicals; Cardiologie -- Périodiques; Cardiology; Electronic journals; Periodicals; 616.12
• DOI: 10.1016/j.yjmcc.2015.04.013
• Languages: English
• ISSNs: 0022-2828

Abstract

Abstract: Reduced phosphorylation of the tumor suppressor p27 Kip1 (p27) at serine 10 (Ser10) is a hallmark of advanced human and mouse atherosclerosis. Apolipoprotein E-null mice defective for this posttranslational modification (apoE −/− p27Ser10Ala) exhibited increased atherosclerosis burden at late disease states. Here, we investigated the regulation of p27 phosphorylation in Ser10 at the very initial stages of atherosclerosis and its impact on endothelial-leukocyte interaction and early plaque formation. Hypercholesterolemia in fat-fed apoE −/− mice is associated with a rapid downregulation of p27-phospho-Ser10 in primary endothelial cells (ECs) and in aorta prior to the development of macroscopically-visible lesions. We find that lack of p27 phosphorylation at Ser10 enhances the expression of adhesion molecules in aorta of apoE −/− mice and ECs, and augments endothelial-leukocyte interactions and leukocyte recruitment in vivo . These effects correlated with increased RhoA/Rho-associated coiled-coil containing protein kinase (ROCK) signaling in ECs, and inhibition of this pathway with fasudil reduced leukocyte-EC interactions to control levels in the microvasculature of p27Ser10Ala mice. Moreover, apoE −/− p27Ser10Ala mice displayed increased leukocyte recruitment and homing to atherosusceptible arteries and augmented early plaque development, which could be blunted with fasudil. In conclusion, our studies demonstrate a very rapid reduction in p27-phospho-Ser10 levels at the onset of atherogenesis, which contributes to early plaque build-up through RhoA/ROCK-induced integrin expression in ECs and enhanced leukocyte recruitment. Graphical abstract: Highlights: p27 is dephosphorylated at Ser10 in aorta of apoE-/- mice before ateroma development. Defective p27-phospho-Ser10 enhances VCAM-1 expression in aorta and endothelium. Defective p27-phopho-Ser10 augments leukocyte recruitment and early atherogenesis. RhoA/ROCK inhibition blunts proatherogenic effect of defective p27-phospho-Ser10.