Proliferative stimulation of the vascular endothelin-1 axis in vitro and ex vivo by infection with Chlamydia pneumoniae

Jan Marco Kern; Viola Maass; Jan Rupp; Matthias Maass

doi:10.1160/TH09-02-0128

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2009; 102(04): 743-753
DOI: 10.1160/TH09-02-0128

Wound Healing and Inflammation/Infection

Schattauer GmbH

Proliferative stimulation of the vascular endothelin-1 axis in vitro and ex vivo by infection with Chlamydia pneumoniae

Authors

Jan Marco Kern

¹Institute of Medical Microbiology, Hygiene and Infectious Diseases, Paracelsus Medical Private University, University Hospital Salzburg, Austria
Viola Maass

¹Institute of Medical Microbiology, Hygiene and Infectious Diseases, Paracelsus Medical Private University, University Hospital Salzburg, Austria
Jan Rupp

²Institute of Medical Microbiology and Hygiene, University Hospital Schleswig-Holstein Campus Luebeck, Luebeck, Germany
Matthias Maass

¹Institute of Medical Microbiology, Hygiene and Infectious Diseases, Paracelsus Medical Private University, University Hospital Salzburg, Austria

Abstract

Summary

Endothelin-1 (ET-1) is a vasoactive peptide that modifies vascular function via the G-protein coupled transmembrane receptors, Endothelin-A receptor (ETAR) and Endothelin-B receptor (ETBR). Dysregulation of the ET-1 axis plays a role in atherosclerotic development as it triggers cell proliferation, inflammation, and vasoconstriction.The respiratory pathogen Chlamydia pneumoniae (Cp) has been recovered from atherosclerotic lesions, and related to atherogenesis, via activation of vascular small GTPases and leukocyte recruitment. Cp effectively reprograms host cell signalling and is able to enter an intracellular persistent state in vascular cells that is refractory to antibiotics. Upon chlamydial infection, vascular smooth muscle cells, which do not produce significant ET-1 under physiological conditions were switched into a fundamental source of ET-1 mRNA and protein in a p38-MAP-kinase-dependent pathway. Endothelial cells did not overproduce ET-1 but showed upregulation of mitogenic ETAR mRNA and protein while the counterbalancing ETBR, which regulates ET-1 clearance, remained unaffected.This disruption of the ET-1 axis was confirmed in an ex vivo mouse aortic ring model, and resulted in endothelial cell proliferation that could be abrogated by ETAR-siRNA and the selective ETAR-antagonist BQ-123. Chronic chlamydial infection of the vascular wall might represent a permanent noxious stimulus linked to the endothelial cell proliferation characteristic of early atherosclerosis. Suppression of this deleterious paracrine loop by ETAR antagonism opens up a new option of preventing possible vascular sequelae of otherwise untreatable chronic chlamydial infection. In conclusion, this is the first study to demonstrate infection to dysregulate the ET-1 axis towards inducing a proatherogenic proliferative phenotype.

Keywords

Chlamydia pneumoniae - Endothelin-1 - Atherosclerosis - Proliferation - Endothelial cells

Full Text

References

References
1 Libby P, Ridker PM, Maseri A. Inflammation and Atherosclerosis. Circulation 2002; 105: 1135-1143.
2 Ross R. Atherosclerosis - an inflammatory disease. N Engl J Med 1999; 340: 115-126.
3 Prasad A, Zhu J, Halcox HPJ, Quyyumi AA. et al. Predisposition to atherosclerosis by infections - Role of endothelial dysfunction. Circulation 2002; 106: 184-190.
4 Maass M, Jahn J, Gieffers J. et al. Detection of Chlamydia pneumoniae within peripheral blood monocytes of patients with unstable angina or myocardial infarction. J Infect Dis 2000; 181: 449-451.
5 Campbell LA, O’Brien ER, Cappucio AL. et al. Detection of Chlamydia pneumoniae (TWAR) in human coronary atherectomy tissues. J Infect Dis 1995; 172: 585-588.
6 Kuo CC, Shor A, Campbell LA. et al. Demonstration of Chlamydia pneumoniae in atherosclerotic lesions of coronary arteries. J Infect Dis 1993; 167: 841-849.
7 Maass M, Bartels C, Engel PM. et al. Endovascular presence of viable Chlamydia pneumoniae is a common phenomenon in coronary artery disease. J Am Coll Cardiol 1998; 187: 103-106.
8 Rupp J, Hellwig-Bürgel T, Wobbe V. et al. Chlamydia pneumoniae infection promotes a proliferative phenotype in the vasculature through Egr-1 activation in vitro and in vivo. Proc Natl Acad Sci USA 2005; 102: 3447-3452.
9 O’Connor CM, Dunne MW, Pfeffer MA. et al. Azithromycin for the secondary prevention of coronary heart disease events: the WIZARD study: a randomised controlled trial. JAMA 2003; 290: 1459-1466.
10 Grayston JT, Kronmal RA, Jackson LA. et al. Azithromycin for the secondary prevention of coronary events. N Engl J Med 2005; 352: 1637-1645.
11 Anderson JL, Muhlestein JB. Antibiotic trials for coronary artery disease. Tex Heart Inst J 2004; 31: 33-38.
12 Jaff MR, Dale RA, Creager MA. et al. Anti- Chlamydial antibiotic therapy for symptom improvement in peripheral artery disease: prospective evaluation of rifalazil effect on vascular symptoms of intermittent claudication and other endpoints in chlamydia pneumoniae seropositive patients (PROVIDENCE-1). Circulation 2009; 119: 452-458.
13 Gieffers J, Füllgraf H, Jahn J. et al. Chlamydia pneumoniae infection in circulating human monocytes is refractory to antibiotic treatment. Circulation 2001; 103: 351-356.
14 Gieffers J, Rupp J, Gebert A. et al. First-choice antibiotics at subinhibitory concentrations induce persistence of Chlamydia pneumoniae. Antimicrob Agents Chemother 2004; 48: 1402-1405.
15 Böhm F, Pernow J. The importance of endothelin-1 for vascular dysfunction in cardiovascular disease. Cardiovasc Res 2007; 76: 8-18.
16 Ivey ME, Osman N, Little PJ. Endothelin-1 signalling in vascular smooth muscle: Pathways controlling cellular associated with atherosclerosis. Atherosclerosis 2008; 199: 237-247.
17 Ishizawa K, Yoshizumi M, Tsuchiya K. et al. Dual effect of Endothelin-1: induction of mesangial cell migration and facilitation of monocyte recruitment through MCP-1 production by mesangial cells. Hypertens Res 2004; 27: 433-440.
18 Lerman A, Edwards BS, Hallett JW. Circulating and tissue endothelin immunoreactivity in advanced atherosclerosis. N Engl J Med 1991; 325: 997-1001.
19 Ohlstein EH, Arleth A, Bryan H. The selective endothelin ETA receptor antagonist BQ 123 antagonizes endothelin-1 mediated mitogenesis. Eur J Pharmacol 1992; 13: 85-88.
20 Li L, Fink GD, Watts SW. et al. Endothelin-1 increases vascular superoxide via endothelin(A)NADPH oxidase pathway in low-renin hypertension. Circulation 2003; 107: 1053-1058.
21 Ak G, Buyukberber S, Sevince A. et al. The relation between plasma endothelin-1 levels and metabolic control, risk factors, treatment modalities, and diabetic microangiopathy in patiens with type 2 diabetes mellitus. J Diabetes Complications 2001; 15: 150-157.
22 Khan IA. Role of endothelin-1 in myocardial infarction. Chest 2005; 127: 1474-1476.
23 Dhaun N, Pollock DM, Goddard J. et al. Selective and mixed endothelin receptor antagonism in cardiovascular disease. Trends Pharmacol Sci 2007; 28: 573-579.
24 Maass M, Harig U. Evaluation of culture conditions used for isolation of Chlamydia pneumoniae. Am J Clin Path 1995; 103: 141-148.
25 Winer J, Jung CS, Shackel I. et al. Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring of gene expression in cardiac myocytes in vitro. Anal Biochem 1999; 270: 41-49.
26 Yang Z, Oemar BS, Carrel T. et al. Different proliferative properties of smooth muscle cells of human arterial and venous bypass vessels: role of PDGF receptors, mitogen-activated protein kinase, and cyclin-dependent kinase inhibitors. Circulation 1998; 97: 181-187.
27 Rupprecht HJ, Blankenberg S, Bickel C. et al. Impact of viral and bacterial infectious burden on longterm prognosis in patients with coronary artery disease. Circulation 2001; 104: 25-31.
28 Shor A, Kuo CC, Patton DL. Detection of Chlamydia pneumoniae in the coronary artery atheroma plaque. South Afr Med J 1992; 82: 158-161.
29 Gaydos CA, Summersgill JT, Sahney NN. et al. Replication of Chlamydia pneumoniae in vitro in human macrophages, endothelial cells, and aortic artery smooth muscle cells. Infect Immun 1996; 64: 1614-1620.
30 Godzik KL, O’Brien ER, Wang AK. et al. In vitro susceptibility of human vascular wall cells to infection with Chlamydia pneumoniae. J Clin Microbiol 1995; 33: 2411-2414.
31 Summersgill JT, Molestina RE, Miller RD, Ramirez JA. Interactions of Chlamydia pneumoniae with human endothelial cells. J Infect Dis 2000; 181: 479-482.
32 Fryer RH, Schwobe EP. Woods et al. Chlamydia species infect human vascular endothelial cells and induce procoagulant activity. J Invest Med 1997; 45: 168-174.
33 Dechend R, Maass M, Gieffers J. et al. Chlamydia pneumoniae infection of vascular smooth muscle and endothelial cells activates NF-κB and induces tissue factor and PAI-1 expression: a potential link to accelerated atherosclerosis. Circulation 1999; 100: 1369-1373.
34 Matsuura A, Yamochi W, Hirata KI. et al. Stimulatory interaction between vascular endothelial growth factor and endothelin-1 on each gene expression. Hypertension 1998; 32: 89-95.
35 Hayasaki Y, Nakajiama M, Kitano Y. et al. ICAM-1 expression on cardiac myocytes and aortic endothelial cells via their specific endothelin receptor subtype. Biochem Biophys Res Comm 1996; 229: 817-824.
36 Hohnemeier H, Pinto YM, Horkay F. et al. Endothelin-converting-enzyme-1 mRNA expression in human cardiovascular diseases. J Cardiovasc Pharmacol 1998; 31: 52-54.
37 Hirata Y, Emori T, Eguchi S. et al. Endothelin receptor subtype B mediates synthesis of nitric oxide by cultured bovine endothelial cells. J Clin Invest 1993; 91: 1367-1373.
38 Shah R. Endothelins in health and disease. Eur J Int Med 2007; 18: 272-282.
39 Woods M, Mitchell JA, Wood EG. et al. Endothelin-1 is induced by cytokines in human vascular smooth muscle cells: evidence for intracellular endothelin-converting enzyme. Mol Pharmacol 1999; 55: 902-909.
40 Zeither AM, Ihling C, Pistorius K. et al. Increased tissue edothelin immunoreactivity in atherosclerotic lesions associated with acute coronary syndromes. Lancet 1994; 344: 1405-1406.
41 Woods M, Wood EG, Bardswell SC. et al. Role for nuclear factor-kappa B and signal transducer and activator of transcription 1/ interferon regulatory factor-1 in cytokine-induced endothelin-1 release in human vascular smooth muscle cells. Mol Pharmacol 2003; 64: 923-931.
42 Zhang WM, Zhou J, Ye QJ. Endothelin-1 enhances proliferation of lung cancer cells by increaseing intracellular free Ca2+. Life Sci 2008; 82: 764-771.
43 Böhm F, Beltran E, Pernow J. Endothelin receptor blockade improves endothelial function in atherosclerotic patients on angiotensin converting enzyme inhibition. J Int Med 2005; 257: 263-271.
44 Halcox JPJ, Nour KRA, Zalos G. et al. Endogenous endothelin in human coronary vascular function. Differential contribution of endothelin receptor types A and B. Hypertension 2007; 49: 1134-1141.
45 Kowala MC, Rose PM, Stein PD. Selective blockade of the endothelin subtype A receptor decreases early atherosclerosis in hamster fed cholesterol. Am J Pathol 1995; 146: 819-826.
46 Barton M, Haudenschild CC, d’Uscio LV. Endothelin ETA receptor blockade restores NO-mediated endothelial function and inhibits atherosclerosis in apoEdeficient mice. Proc Natl Acad Sci USA 1998; 95: 14367-14372.
47 Kern JM, Maass V, Maass M. Molecular pathogenesis of chronic Chlamydia pneumoniae infection: a brief overview. Clin Microbiol Infect 2009; 15: 36-41.