Known players, new interplay in atherogenesis: Chronic shear stress and carbamylated-LDL induce and modulate expression of atherogenic LR11 in human coronary artery endothelium

Tarek M. Bajar; Wolfgang Winnicki; Eva-Theres Gensberger; Susanna I. Scharrer; Heinz Regele; Klaus Aumayr; Chantal Kopecky; Bernhard M. Gmeiner; Marcela Hermann; Robert Zeillinger; Gürkan Sengölge

doi:10.1160/TH12-12-0924

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2014; 111(02): 323-332
DOI: 10.1160/TH12-12-0924

Cardiovascular Biology and Cell Signalling

Schattauer GmbH

Known players, new interplay in atherogenesis: Chronic shear stress and carbamylated-LDL induce and modulate expression of atherogenic LR11 in human coronary artery endothelium

Authors

Tarek M. Bajar^*

¹Department of Medicine III, Division of Nephrology and Dialysis, Medical University of Vienna, Vienna, Austria
Wolfgang Winnicki^*

¹Department of Medicine III, Division of Nephrology and Dialysis, Medical University of Vienna, Vienna, Austria
Eva-Theres Gensberger

²Department of Medical Biochemistry, Max F. Perutz Laboratories, Medical University of Vienna, Vienna, Austria
Susanna I. Scharrer

¹Department of Medicine III, Division of Nephrology and Dialysis, Medical University of Vienna, Vienna, Austria
Heinz Regele

³Institute of Clinical Pathology, Medical University of Vienna, Vienna, Austria
Klaus Aumayr

³Institute of Clinical Pathology, Medical University of Vienna, Vienna, Austria
Chantal Kopecky

¹Department of Medicine III, Division of Nephrology and Dialysis, Medical University of Vienna, Vienna, Austria
Bernhard M. Gmeiner

⁴Department of Medical Chemistry and Pathobio-chemistry, Medical University of Vienna, Vienna, Austria
Marcela Hermann

²Department of Medical Biochemistry, Max F. Perutz Laboratories, Medical University of Vienna, Vienna, Austria
Robert Zeillinger

⁵Department of Obstetrics and Gynecology and Ludwig Boltzmann Cluster Translational Oncology, Medical University of Vienna, Vienna, Austria
Gürkan Sengölge

⁴Department of Medical Chemistry and Pathobio-chemistry, Medical University of Vienna, Vienna, Austria

Abstract

Summary

In this study we examined whether low-density lipoprotein (LDL) receptor family members represent a link between blood flow characteristics and modified low-density lipoproteins involved in endothelial injury, a pivotal factor in atherogenesis. We demonstrated the expression of pro-atherogenic LDL receptor relative (LR11) for the first time in human coronary artery endothelial cells (HCAEC) in vitro and in vivo. Next, LR11 expression and regulation were explored in HCAEC cultured conventionally or on the inner surface of hollow fiber capillaries under exposure to shear stress for 10 days in the presence or absence of LDL. There was no LR11 expression under static conditions. When exposed to chronic low shear stress (2.5 dynes/cm²) transmembrane and soluble endothelial-LR11 were detected in high levels irrespective of the type of LDL added (carbamylated or native). In contrast, chronic high shear stress (25 dynes/cm²) inhibited the LR11-inducing effect of LDL such that transmembrane and soluble LR11 expression became non-detectable with native LDL. Carbamylated LDL significantly counteracted this atheroprotective effect of high shear stress as shown by lower, yet sustained expression of soluble and transmembrane LR11. Oxidised LDL showed similar effects compared to carbamylated LDL but caused significantly lower LR11 expression under chronic high shear stress. Medium from HCAEC under LR11-inducing conditions enhanced vascular smooth muscle cell migration, which was abrogated by the anti-LR11 antibody. Expression of LR11 depended entirely on p38MAPK phosphorylation. We conclude that coronary endothelial LR11 expression modulated by LDL and chronic shear stress contributes to atherogenesis. LR11 and p38MAPK are potential targets for prevention of atherosclerosis.

Keywords

Atherosclerosis - endothelium - modified lipoproteins - shear stress - smooth muscle cell

Full Text

References

References
1 Apostolov EO. et al. Chronic uremia stimulates LDL carbamylation and atherosclerosis. J Am Soc Nephrol 2010; 21: 1852-1857.
2 Apostolov EO. et al. Carbamylated low-density lipoprotein induces monocyte adhesion to endothelial cells through intercellular adhesion molecule-1 and vascular cell adhesion molecule-1. Arterioscler Thromb Vasc Biol 2007; 27: 826-832.
3 Gleissner CA. et al. Effects of native and modified low-density lipoproteins on monocyte recruitment in atherosclerosis. Hypertension 2007; 50: 276-283.
4 Ok E. et al. Carbamylated low-density lipoprotein induces death of endothelial cells: a link to atherosclerosis in patients with kidney disease. Kidney Int 2005; 68: 173-178.
5 Ku DN. et al. Pulsatile flow and atherosclerosis in the human carotid bifurcation. Positive correlation between plaque location and low oscillating shear stress. Arteriosclerosis 1985; 5: 293-302.
6 Chatzizisis YS. et al. Role of endothelial shear stress in the natural history of coronary atherosclerosis and vascular remodeling: molecular, cellular, and vascular behavior. J Am Coll Cardiol 2007; 49: 2379-2393.
7 Wang Z. et al. Protein carbamylation links inflammation, smoking, uremia and atherogenesis. Nat Med 2007; 13: 1176-1184.
8 Weiner DE. et al. Chronic kidney disease as a risk factor for cardiovascular disease and all-cause mortality: a pooled analysis of community-based studies. J Am Soc Nephrol 2004; 15: 1307-1315.
9 Horiuchi S. et al. Scavenger receptors for oxidized and glycated proteins. Amino Acids 2003; 25: 283-292.
10 Asci G. et al. Carbamylated low-density lipoprotein induces proliferation and increases adhesion molecule expression of human coronary artery smooth muscle cells. Nephrology 2008; 13: 480-486.
11 Basnakian AG. et al. Carbamylated LDL. Adv Clin Chem 2010; 51: 25-52.
12 Shah SV. et al. Novel mechanisms in accelerated atherosclerosis in kidney disease. J Ren Nutr 2008; 18: 65-69.
13 Kanaki T. et al. Expression of LR11, a mosaic LDL receptor family member, is markedly increased in atherosclerotic lesions. Arterioscler Thromb Vasc Biol 1999; 19: 2687-2695.
14 Tanaga K. et al. LRP1B attenuates the migration of smooth muscle cells by reducing membrane localization of urokinase and PDGF receptors. Arterioscler Thromb Vasc Biol 2004; 24: 1422-1428.
15 Zhu Y. et al. LR11, an LDL receptor gene family member, is a novel regulator of smooth muscle cell migration. Circ Res 2004; 94: 752-758.
16 Morwald S. et al. A novel mosaic protein containing LDL receptor elements is highly conserved in humans and chickens. Arterioscler Thromb Vasc Biol 1997; 17: 996-1002.
17 Böhm C. et al. SorLA signaling by regulated intramembrane proteolysis. J Biol Chem 2006; 281: 14547-14553.
18 Ohwaki K. et al. A secreted soluble form of LR11, specifically expressed in inti-mal smooth muscle cells, accelerates formation of lipid-laden macrophages. Ar- terioscler Thromb Vasc Biol 2007; 27: 1050-1056.
19 Matsuo M. et al. Development of an immunoassay for the quantification of soluble LR11, a circulating marker of atherosclerosis. Clin Chem 2009; 55: 1801-1808.
20 Takahashi M. et al. Enhanced circulating soluble LR11 in patients with coronary organic stenosis. Atherosclerosis 2010; 210: 581-584.
21 Burkitt MJ. A critical overview of the chemistry of copper-dependent low density lipoprotein oxidation: roles of lipid hydroperoxides, alpha-tocopherol, thiols, and ceruloplasmin. Arch Biochem Biophys 2001; 394: 117-135.
22 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using realtime quantitative PCR and the 2 AACt Method. Methods 2001; 25: 402-408.
23 Bajari TM. et al. A model for modulation of leptin activity by association with clusterin. Faseb J 2003; 17: 1505-1507.
24 Poling J. et al. Induction of smooth muscle cell migration during arteriogenesis is mediated by rap2. Arterioscler Thromb Vasc Biol 2011; 31: 2297-2305.
25 Schreier B. et al. Consequences of epidermal growth factor receptor (ErbB1) loss for vascular smooth muscle cells from mice with targeted deletion of ErbB1. Arterioscler Thromb Vasc Biol 2011; 31: 1643-1652.
26 Duran-Prado M. et al. Cortistatin inhibits migration and proliferation of human vascular smooth muscle cells and decreases neointimal formation on carotid artery ligation. Circ Res 2013; 112: 1444-1455.
27 Shaik SS. et al. Low intensity shear stress increases endothelial ELR+ CXC che-mokine production via a focal adhesion kinase-p38 β MAPK-NF-kB pathway. J Biol Chem 2009; 284: 5945-5955.
28 Wang WY. et al. OxLDL stimulates lipoprotein-associated phospholipase A2 expression in THP-1 monocytes via PI3K and p38 MAPK pathways. Cardiovasc Res 2010; 85: 845-852.
29 Ettelaie C. et al. p38alpha phosphorylates serine 258 within the cytoplasmic domain of tissue factor and prevents its incorporation into cell-derived micropar-ticles. Biochim Biophys Acta 2013; 1833: 613-621.
30 Petry A. et al. NOX2 and NOX4 mediate proliferative response in endothelial cells. Antioxid Redox Signal 2006; 8: 1473-1484.
31 Zhu Y. et al. Enhanced expression of the LDL receptor family member LR11 increases migration of smooth muscle cells in vitro. Circulation 2002; 105: 1830-1836.
32 Malek A, Izumo S. Physiological fluid shear stress causes downregulation of en-dothelin-1 mRNA in bovine aortic endothelium. Am J Physiol 1992; 263: C389-396.
33 Chiu JJ. et al. Shear stress increases ICAM-1 and decreases VCAM-1 and E-se-lectin expressions induced by tumor necrosis factor-[alpha] in endothelial cells. Arterioscler Thromb Vasc Biol 2004; 24: 73-79.
34 Cheng C. et al. Shear stress-induced changes in atherosclerotic plaque composition are modulated by chemokines. J Clin Invest 2007; 117: 616-626.
35 de Nigris F. et al. Beneficial effects of antioxidants and L-arginine on oxidation-sensitive gene expression and endothelial NO synthase activity at sites of disturbed shear stress. Proc Natl Acad Sci USA 2003; 100: 1420-1425.
36 de Nigris F. et al. Beneficial effects of pomegranate juice on oxidation-sensitive genes and endothelial nitric oxide synthase activity at sites of perturbed shear stress. Proc Natl Acad Sci USA 2005; 102: 4896-4901.
37 Won D. et al. Relative reduction of endothelial nitric-oxide synthase expression and transcription in atherosclerosis-prone regions of the mouse aorta and in an in vitro model of disturbed flow. Am J Pathol 2007; 171: 1691-1704.
38 Warboys CM. et al. Acute and chronic exposure to shear stress have opposite effects on endothelial permeability to macromolecules. Am J Physiol Heart Circ Physiol 2010; 298: H1850-1856.
39 Apostolov EO. et al. Carbamylated low-density lipoprotein: nontraditional risk factor for cardiovascular events in patients with chronic kidney disease. J Renal Nutr 2012; 22: 134-138.
40 Apostolov EO. et al. Scavenger receptors of endothelial cells mediate the uptake and cellular proatherogenic effects of carbamylated LDL. Arterioscl Thromb Vasc Biol 2009; 29: 1622-1630.
41 Dai G. et al. Distinct endothelial phenotypes evoked by arterial waveforms derived from atherosclerosis-susceptible and -resistant regions of human vascula-ture. Proc Natl Acad Sci USA 2004; 101: 14871-14876.
42 Illi B. et al. Shear stress-mediated chromatin remodeling provides molecular basis for flow-dependent regulation of gene expression. Circulation Res 2003; 93: 155-161.
43 Breton-Romero R. et al. Critical role of hydrogen peroxide signaling in the sequential activation of p38 MAPK and eNOS in laminar shear stress. Free Rad Biol Med 2012; 52: 1093-1100.
44 Ronco C. et al. Cardiorenal syndrome. J Am Coll Cardiol 2008; 52: 1527-1539.
45 Verbeke FH. et al. Local shear stress and brachial artery functions in end-stage renal disease. J Am Soc Nephrol 2007; 18: 621-628.

Supplementary Material

Online Supplementary Material (PDF) (PDF)