Oliver Borst1, Malte Schaub1, Britta Walker1, Evi Schmid1, Patrick Münzer1, Jakob Voelkl1, Ioana Alesutan1, José M Rodríguez1, Sebastian Vogel1, Tanja Schoenberger1, Katja Metzger1, Dominik Rath1, Anja Umbach1, Dietmar Kuhl1, Iris I Müller1, Peter Seizer1, Tobias Geisler1, Meinrad Gawaz1, Florian Lang2. 1. From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.). 2. From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.). florian.lang@uni-tuebingen.de meinrad.gawaz@med.uni-tuebingen.de.
Abstract
OBJECTIVE: Atherosclerosis, an inflammatory disease of arterial vessel walls, requires migration and matrix metalloproteinase (MMP)-9-dependent invasion of monocytes/macrophages into the vascular wall. MMP-9 expression is stimulated by transcription factor nuclear factor-κB, which is regulated by inhibitor κB (IκB) and thus IκB kinase. Regulators of nuclear factor-κB include serum- and glucocorticoid-inducible kinase 1 (SGK1). The present study explored involvement of SGK1 in vascular inflammation and atherogenesis. APPROACH AND RESULTS: Gene-targeted apolipoprotein E (ApoE)-deficient mice without (apoe(-/-)sgk1(+/+)) or with (apoe(-/-)sgk1(-/-)) additional SGK1 knockout received 16-week cholesterol-rich diet. According to immunohistochemistry atherosclerotic lesions in aorta and carotid artery, vascular CD45(+) leukocyte infiltration, Mac-3(+) macrophage infiltration, vascular smooth muscle cell content, MMP-2, and MMP-9 positive areas in atherosclerotic tissue were significantly less in apoe(-/-)sgk1(-/-)mice than in apoe(-/-)sgk1(+/+)mice. As determined by Boyden chamber, thioglycollate-induced peritonitis and air pouch model, migration of SGK1-deficient CD11b(+)F4/80(+) macrophages was significantly diminished in vitro and in vivo. Zymographic MMP-2 and MMP-9 production, MMP-9 activity and invasion through matrigel in vitro were significantly less in sgk1(-/-) than in sgk1(+/+)macrophages and in control plasmid-transfected or inactive (K127N)SGK1-transfected than in constitutively active (S422D)SGK1-transfected THP-1 cells. Confocal microscopy revealed reduced macrophage number and macrophage MMP-9 content in plaques of apoe(-/-)sgk1(-/-) mice. In THP-1 cells, MMP-inhibitor GM6001 (25 μmol/L) abrogated (S422D)SGK1-induced MMP-9 production and invasion. According to reverse transcription polymerase chain reaction, MMP-9 transcript levels were significantly reduced in sgk1(-/-)macrophages and strongly upregulated in (S422D)SGK1-transfected THP-1 cells compared with control plasmid-transfected or (K127N)SGK1-transfected THP-1 cells. According to immunoblotting and confocal microscopy, phosphorylation of IκB kinase and inhibitor κB and nuclear translocation of p50 were significantly lower in sgk1(-/-)macrophages than in sgk1(+/+)macrophages and significantly higher in (S422D)SGK1-transfected THP-1 cells than in control plasmid-transfected or (K127N)SGK1-transfected THP-1 cells. Treatment of (S422D)SGK1-transfected THP-1 cells with IκB kinase-inhibitor BMS-345541 (10 μmol/L) abolished (S422D)SGK1-induced increase of MMP-9 transcription and gelatinase activity. CONCLUSIONS: SGK1 plays a pivotal role in vascular inflammation during atherogenesis. SGK1 participates in the regulation of monocyte/macrophage migration and MMP-9 transcription via regulation of nuclear factor-κB.
OBJECTIVE:Atherosclerosis, an inflammatory disease of arterial vessel walls, requires migration and matrix metalloproteinase (MMP)-9-dependent invasion of monocytes/macrophages into the vascular wall. MMP-9 expression is stimulated by transcription factor nuclear factor-κB, which is regulated by inhibitor κB (IκB) and thus IκB kinase. Regulators of nuclear factor-κB include serum- and glucocorticoid-inducible kinase 1 (SGK1). The present study explored involvement of SGK1 in vascular inflammation and atherogenesis. APPROACH AND RESULTS: Gene-targeted apolipoprotein E (ApoE)-deficient mice without (apoe(-/-)sgk1(+/+)) or with (apoe(-/-)sgk1(-/-)) additional SGK1 knockout received 16-week cholesterol-rich diet. According to immunohistochemistry atherosclerotic lesions in aorta and carotid artery, vascular CD45(+) leukocyte infiltration, Mac-3(+) macrophage infiltration, vascular smooth muscle cell content, MMP-2, and MMP-9 positive areas in atherosclerotic tissue were significantly less in apoe(-/-)sgk1(-/-)mice than in apoe(-/-)sgk1(+/+)mice. As determined by Boyden chamber, thioglycollate-induced peritonitis and air pouch model, migration of SGK1-deficientCD11b(+)F4/80(+) macrophages was significantly diminished in vitro and in vivo. Zymographic MMP-2 and MMP-9 production, MMP-9 activity and invasion through matrigel in vitro were significantly less in sgk1(-/-) than in sgk1(+/+)macrophages and in control plasmid-transfected or inactive (K127N)SGK1-transfected than in constitutively active (S422D)SGK1-transfected THP-1 cells. Confocal microscopy revealed reduced macrophage number and macrophage MMP-9 content in plaques of apoe(-/-)sgk1(-/-) mice. In THP-1 cells, MMP-inhibitor GM6001 (25 μmol/L) abrogated (S422D)SGK1-induced MMP-9 production and invasion. According to reverse transcription polymerase chain reaction, MMP-9 transcript levels were significantly reduced in sgk1(-/-)macrophages and strongly upregulated in (S422D)SGK1-transfected THP-1 cells compared with control plasmid-transfected or (K127N)SGK1-transfected THP-1 cells. According to immunoblotting and confocal microscopy, phosphorylation of IκB kinase and inhibitor κB and nuclear translocation of p50 were significantly lower in sgk1(-/-)macrophages than in sgk1(+/+)macrophages and significantly higher in (S422D)SGK1-transfected THP-1 cells than in control plasmid-transfected or (K127N)SGK1-transfected THP-1 cells. Treatment of (S422D)SGK1-transfected THP-1 cells with IκB kinase-inhibitor BMS-345541 (10 μmol/L) abolished (S422D)SGK1-induced increase of MMP-9 transcription and gelatinase activity. CONCLUSIONS:SGK1 plays a pivotal role in vascular inflammation during atherogenesis. SGK1 participates in the regulation of monocyte/macrophage migration and MMP-9 transcription via regulation of nuclear factor-κB.
Authors: Saskia N I von Ungern-Sternberg; Sebastian Vogel; Britta Walker-Allgaier; Sascha Geue; Andreas Maurer; Anna-Maria Wild; Patrick Münzer; Madhumita Chatterjee; David Heinzmann; Elisabeth Kremmer; Oliver Borst; Patricia Loughran; Alma Zernecke; Matthew D Neal; Timothy R Billiar; Meinrad Gawaz; Peter Seizer Journal: Thromb Haemost Date: 2017-11-30 Impact factor: 5.249
Authors: Mariángeles Clauzure; Ángel G Valdivieso; Andrea V Dugour; Consuelo Mori; María M Massip-Copiz; María Á Aguilar; Verónica Sotomayor; Cristian J A Asensio; Juan M Figueroa; Tomás A Santa-Coloma Journal: Immunology Date: 2021-05-02 Impact factor: 7.215
Authors: M Chatterjee; S N I von Ungern-Sternberg; P Seizer; F Schlegel; M Büttcher; N A Sindhu; S Müller; A Mack; M Gawaz Journal: Cell Death Dis Date: 2015-11-19 Impact factor: 8.469