Extracellular RNA promotes leukocyte recruitment in the vascular system by mobilising proinflammatory cytokines

Silvia Fischer; Tobias Grantzow; Judith I. Pagel; Marlene Tschernatsch; Markus Sperandio; Klaus T. Preissner; Elisabeth Deindl

doi:10.1160/TH12-03-0186

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2012; 108(04): 730-741
DOI: 10.1160/TH12-03-0186

Wound Healing and Inflammation/Infection

Schattauer GmbH

Extracellular RNA promotes leukocyte recruitment in the vascular system by mobilising proinflammatory cytokines

Silvia Fischer^*

¹Department of Biochemistry, Medical School, Justus-Liebig-University, Giessen, Germany

,

Tobias Grantzow^*

²Walter-Brendel-Centre of Experimental Medicine, Ludwig-Maximilians-University, Munich, Germany

,

Judith I. Pagel

²Walter-Brendel-Centre of Experimental Medicine, Ludwig-Maximilians-University, Munich, Germany

,

Marlene Tschernatsch

³Department of Neurology, Justus-Liebig-University Giessen, Giessen

,

Markus Sperandio

²Walter-Brendel-Centre of Experimental Medicine, Ludwig-Maximilians-University, Munich, Germany

,

Klaus T. Preissner

¹Department of Biochemistry, Medical School, Justus-Liebig-University, Giessen, Germany

,

Elisabeth Deindl

²Walter-Brendel-Centre of Experimental Medicine, Ludwig-Maximilians-University, Munich, Germany

› Author Affiliations

Abstract

Summary

Extracellular RNA (eRNA), released from cells under conditions of injury or vascular disease, acts as potent prothrombotic factor and promotes vascular hyperpermeability related to oedema formation in vivo. In this study, we aimed to investigate the mechanism by which eRNA triggers inflammatory processes, particularly associated with different steps of leukocyte recruitment. Using intravital microscopy of murine cremaster muscle venules, eRNA (but not DNA) significantly induced leukocyte adhesion and transmigration in vivo, which was comparable in its effects to the function of tumour-necrosis-factor-α (TNF-α). In vitro, eRNA promoted adhesion and transmigration of monocytic cells on and across endothelial cell monolayers. eRNA-induced monocyte adhesion in vitro was mediated by activation of the vascular endothelial growth factor (VEGF)/VEGF-receptor-2 system and was abolished by neutralising antibodies against intercellular adhesion molecule-1 or the p2-inte-grin Mac-1. Additionally, eRNA induced the release of TNF-α from monocytic cells in a time- and concentration-dependent manner, which involved activation of TNF- α -converting enzyme (TACE) as well as the nuclear factor kB signalling machinery. In vivo, inhibiton of TACE significantly reduced eRNA-induced leukocyte adhesion. Our findings present evidence that eRNA in connection with tissue/vascular damage provokes a potent inflammatory response by inducing leukocyte recruitment and by mobilising proinflammatory cytokines from monocytes.

Keywords

Extracellular RNA - inflammation - leukocyte recruitment - TNF-α - endothelial cells - vasculature

Full Text

References

References
1 Wieczorek AJ, Rhyner C, Block LH. Isolation and characterization of an RNA-proteolipid complex assiciated with the malignant state in humans. Proc Natl Acad Sci USA 1985; 82: 3455-3459.
2 Garcia-Olmo DC, Ruiz-Piqueras R, Garcia-Olmo D. Circulating nucleic acids in plasma and serum (CNAPS) and its relation to stem cells and cancer metastasis: state of the issue. Histol Histopathol 2004; 19: 575-583.
3 Kannemeier C, Shibamiya A, Nakazawa F. et al. Extracellular RNA constitutes a natural procoagulant cofactor in blood coagulation. Proc Natl Acad Sci USA 2007; 104: 6388-6393.
4 Fischer S, Gerriets T, Wessels C. et al. Extracellular RNA mediates endothelial-cell permeability via vascular endothelial growth factor. Blood 2007; 110: 2457-2465.
5 Fischer S, Nishio M, Peters SC. et al. Signaling mechanism of extracellular RNA in endothelial cells. FASEB J 2009; 23: 2100-2109.
6 Landre JBP, Hewett PW, Olivot J-M. et al. Human endothelial cells selectively express large amounts of pancreatic-type ribonuclease (RNase 1). J Cell Biochem 2002; 86: 540-552.
7 Moenner M, Hatzi E, Bader J. Secretion of ribonucleases by normal and immortalized cells grown in serum-free culture conditions. In Vitro Cell Dev Biol Anim 1997; 33: 553-561.
8 Weickmann JL, Olson EM, Glitz DG. Immunological assay of pancreatic ribonuclease in serum as an indicator of pancreatic cancer. Cancer Res 1984; 44: 1682-1687.
9 Fischer S, Nishio M, Dadkhahi S. et al. Expression and localisation of vascular ribonucleases in endothelial cells. Thromb Haemost 2011; 105: 345-355.
10 Sperandio M, Frommhold D, Babushkina I. et al. Alpha 2,3-sialyltransferase-IV is essential for L-selectin ligand function in inflammation. Eur J Immunol 2006; 36: 3207-3215.
11 Frommhold D, Kamphues A, Hepper I. et al. RAGE and ICAM-1 cooperate in mediating leukocyte recruitment during acute inflammation in vivo. Blood 2010; 116: 841-849.
12 Weksler BB, Subileau Perrie’re EA. et al. Blood-brain barrier-specific properties of a human adult brain endothelial cell line. Faseb J 2005; 19: 1872-1874.
13 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidi-nium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-159.
14 Black RA, Rauch CT, Kozlosky CJ. et al. A metalloproteinase disintegrin that releases tumor-necrosis factor-alpha from cells. Nature 1997; 385: 729-733.
15 Tsui NB, Ng EK, Lo YM. Stability of endogenous and added RNA in blood specimens, serum, and plasma. Clin Chem 2002; 48: 1647-1653.
16 Kopreski MS, Benko FA, Gocke CD. Circulating RNA as a tumor marker: detection of 5T4 mRNA in breast and lung cancer patient serum. Ann NY Acad Sci 2001; 945: 172-178.
17 Reddi KK, Holland JF. Elevated serum ribonuclease in patients with pancreatic cancer. Proc Natl Acad Sci USA 1976; 73: 2308-2310.
18 Rosi A, Guidoni L, Luciani AM. et al. RNA-lipid complexes released from the plasma emembrane of human colon carcinoma cells. Cancer Lett 1988; 39: 153-160.
19 Sisco KL. Is RNA in serum bound to nucleoprotein complexes?. Clin Chem 2000; 47: 1744-1745.
20 Stroun M, Anker P, Beljanski M. et al. Presence of RNA in the nucleoprotein complex spontaneously released by human lymphocytes and frog auricles in culture. Cancer Res 1978; 38: 3546-3554.
21 Ekström K, Valadi H, Sjöstrand M. et al. Characterization of mRNA and microRNA in human mast cell-derived exosomes and their transfer to other mast cells and blood CD34 progenitor cells. J Extracell Vesicles 2012; 01: 1-12.
22 Clauss M, Weich H, Breier G. et al. The Vascular Endothelial Growth Factor Receptor Flt-1 mediates biological activities: Implications for a functional role of Placenta Growth Factor in monocyte activation and chemotaxis. J Biol Chem 1996; 271: 17629-17634.
23 Akuzawa N, Kurabayashi M, Ohyama Y. et al. Zinc finger transcripton factor Egr-1 activates Flt-1 gene expression in THP-1 cells on induction for macrophage differentiation. Arterioscler Thromb Vasc Biol 2000; 20: 377-384.
24 Heil M, Clauss M, Suzuki K. et al. Vascular endothelial growth factor (VEGF) stimulates monocyte migration through endothelial monolayers via increased integrin expression. Europ J Cell Biol 2000; 79: 850-857.
25 Buscher K, Riese SB, Shakibaei M. et al. The transmembrane domains of L-selectin and CD44 regulate receptor cell surface positioning and leukocyte adhesion under flow. J Biol Chem 2010; 285: 13490-13497.
26 Melder RJ, Yuan J, Munn LL. et al. Erythrocytes enhance lymphocyte rolling and arrest in vivo. Microvasc Res 2000; 59: 316-322.
27 Gupta M, Mahanty S, Ahmed R. et al. Monocyte-derived human macrophages and peripheral blood mononuclear cells infected with ebola virus secrete MIP-1alpha and TNF-alpha and inhibit poly-IC-induced IFN-alpha in vitro. Virology 2001; 284: 20-25.
28 Serbina NV, Cherny M, Shi C. et al. Distinct responses of human monocyte subsets to Aspergillus fumigatus conidia. J Immunol 2009; 183: 2678-2687.
29 Abraham E. NF-kappaB activation. Crit Care Med 2000; 28: N100-N104.
30 Jung YJ, Isaacs JS, Lee S. et al. Il-1beta-mediated up-regulation of HIF-1alpha via an NFkappaB/COX-2 pathway identifies HIF-1 as a critical link between inflammation an oncogenesis. FASEB J 2003; 17: 2115-2117.
31 Scott AJ, O'Dea KP, O'Callaghan D. et al. Reactive oxygen species and p38 MAPK mediate TNF-alpha converting enzyme (TACE/ADAM17) activation in primary human monocytes. J Biol Chem 2011; 24: 1-13.
32 Maltzan K, Tan W, Pruett SB. Investigation of the role of TNF-α converting enzyme (TACE) in the inhibition of cell surface and soluble TNF-α production by acute ethanol exposure. Plos ONE 2012; 07: e29890.
33 Domonkos A, Udvardy A, Lazlo L. et al. Receptor-like properties of the 26 kDa transmembrane form of TNF. Eur Cytokine Netw 2001; 12: 411-419.
34 Peschon JJ, Slack JL, Reddy P. et al. An essential role for ectodomain shedding in mammalian development. Science 1998; 282: 1281-1284.
35 Fan H, Derynck R. Ectodomain shedding of TGF-alpha and other transmembrane proteins is induced by receptor tyrosine kinase activation and MAP kinase signaling cascades. EMBO J 1999; 18: 6962-6972.