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Thromb Haemost 2011; 105(02): 345-355
DOI: 10.1160/TH10-06-0345
Endothelium and Vascular Development
Schattauer GmbH

Expression and localisation of vascular ribonucleases in endothelial cells

Silvia Fischer   *
1   Department of Biochemistry, Justus-Liebig-University, Giessen, Germany
,
Miwako Nishio   *
2   Department of Health Care Sciences, Tokyo Medical and Dental University, Tokyo, Japan
,
Sara Dadkhahi
1   Department of Biochemistry, Justus-Liebig-University, Giessen, Germany
,
Julia Gansler
1   Department of Biochemistry, Justus-Liebig-University, Giessen, Germany
,
Mona Saffarzadeh
1   Department of Biochemistry, Justus-Liebig-University, Giessen, Germany
,
Aya Shibamiyama
1   Department of Biochemistry, Justus-Liebig-University, Giessen, Germany
,
Nicolé Kral
1   Department of Biochemistry, Justus-Liebig-University, Giessen, Germany
,
Nelli Baal
3   Department of Immunology and Transfusion Medicine, Justus-Liebig-University, Giessen, Germany
,
Takatoshi Koyama
2   Department of Health Care Sciences, Tokyo Medical and Dental University, Tokyo, Japan
,
Elisabeth Deindl
4   Walter-Brendel-Centre of Experimental Medicine, Ludwig-Maximilians-University, Munich, Germany
,
Klaus T. Preissner
1   Department of Biochemistry, Justus-Liebig-University, Giessen, Germany
› Author Affiliations Financial support: Studies were generously supported by grants (SFB 547, SPP 1190, FI 543/2–1) and the Excellence cluster “Cardio-pulmonary System” (ECCPS) and the IRTG-1566 (PROMISE) from the Deutsche Forschungsgemeinschaft (DFG, Bonn, Germany) received by Klaus T. Preissner and Silvia Fischer.
Further Information

Publication History

Received: 08 June 2010

Accepted after major revision: 05 November 2010

Publication Date:
25 November 2017 (online)

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Summary

The functions of extracellular RNA in the vascular system as new procoagulatory and permeability-increasing factor in vivoand in vitrowere shown to be counteracted by pancreatic type RNase1. Based on the identification of RNase1 in plasma and serum, it is proposed that the enzyme is expressed by vascular cells to contribute in the regulation of extracellular RNA. It is demonstrated that RNase1 and RNase5 (also termed angiogenin) were differentially expressed in various types of endothelial cells, whereby human umbilical vein endothelial cells (HUVEC) expressed and released the highest concentration of active RNase1. Expression and release of RNase5 were similar in all types of endothelial cells tested. Both RNases were constitutively produced and secreted, whereby a portion of RNase1, but not RNase5, was stored in Weibel-Palade bodies, co-localising with von Willlebrand factor and P-selectin. Accordingly, immediate release of RNase1 from these granules was demonstrated in vitroand in vivousing Weibel-Palade body exocytosis-inducing agents. Additionally, extracellular RNA or poly:IC (but not DNA) induced this short-term release of RNase1. Our results indicate that vascular RNase1 and RNase5 are mainly produced by vascular endothelial cells and can serve, depending on the vascular bed, different functions in vascular homeostasis and endothelial cell responses.

Keywords

Human pancreatic RNase1 - angiogenin - endothelial cells - WeibelPalade bodies

* These authors contributed equally to this study.


 

  • References

  • 1 Beintema JJ, Wietzes P, Weickmann JL. et al. The amino acid sequence of human pancreatic ribonuclease. Anal Biochem 1984; 136: 48-64.
    CrossrefPubMedSearch in Google Scholar
  • 2 Weickmann JL, Elson M, Glitz DG. Purification and characterization of human pancreatic ribonuclease. Biochemistry 1981; 20: 1272-1278.
    CrossrefPubMedSearch in Google Scholar
  • 3 Ribo M, Benito A, Canals A. et al. Purification of engineered human pancreatic ribonuclease. Methods Enzymol 2001; 341: 221-234.
    PubMedSearch in Google Scholar
  • 4 Beintema JJ, Hofsteenge J, Iwana M. et al. Amino acid sequence of the nonsecre-tory ribonuclease of human urine. Biochemistry 1988; 27: 4530-4538.
    CrossrefPubMedSearch in Google Scholar
  • 5 Bläser J, Triebel S, Kopp C. et al. A highly sensitive immunoenzymometric assay for the determination of angiogenin. Eur J Clin Chem Clin Biochem 1993; 31: 513-516.
    PubMedSearch in Google Scholar
  • 6 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.
    PubMedSearch in Google Scholar
  • 7 Leland PA, Staniszewski KE, Park C. et al. The ribonucleolytic activity of angiogenin. Biochemistry 2002; 41: 1343-1350.
    CrossrefPubMedSearch in Google Scholar
  • 8 Fett JW, Bethune JL, Vallee BL. Induction of angiogenesis by mixtures of two angiogenic proteins, angiogenin and acidic fibroblast growth factor, in the chicken chorioallantoic membrane. Biochem Biophys Res Commun 1987; 146: 1122-1131.
    CrossrefPubMedSearch in Google Scholar
  • 9 Fett JW, Strydom DJ, Lobb RR. et al. Isolation and characterization of angiogenin, an angiogenic protein from human carcinoma cells. Biochemistry 1985; 24: 5480-5486.
    CrossrefPubMedSearch in Google Scholar
  • 10 Hu GF, Riordan JF, Vallee BL. A putative angiogenin receptor in angiogenin-responsive human endothelial cells. Proc Natl Acad Sci USA 1997; 94: 2204-2209.
    CrossrefPubMedSearch in Google Scholar
  • 11 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.
    CrossrefPubMedSearch in Google Scholar
  • 12 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.
    CrossrefPubMedSearch in Google Scholar
  • 13 Petrova TV, Mäkinen TPM, Saarela J. et al. Lymphatic endothelial reprogramming of vascular endothelial cells by the Prox-1 homeobox transcription factor. EMBO J 2002; 21: 4593-4599.
    CrossrefPubMedSearch in Google Scholar
  • 14 Futami J, Tsushima Y, Murato Y. et al. Tissue-specific expression of pancreatic-type RNases and RNase inhibitor in humans. DNA Cell Biol 1997; 16: 413-419.
    CrossrefPubMedSearch in Google Scholar
  • 15 Sasso MP, Lombardi M, Confalone E. et al. The differential pattern of tissue-specific expression of ruminant pancreatic type ribonucleases may help to understand the evolutionary history of their genes. Gene 1999; 227: 205-212.
    CrossrefPubMedSearch in Google Scholar
  • 16 Beintema JJ. The primary structure of langur (Presbytis entellus) pancreatic ribonuclease: adaptive features in digestive enzymes in mammals. Mol Biol Evol 1990; 7: 470-477.
    PubMedSearch in Google Scholar
  • 17 Libonati M, Sorrentino S. Degradation of double-stranded RNA by mammalian pancreatic-type ribonucleases. Methods Enzymol 2001; 341: 234-248.
    PubMedSearch in Google Scholar
  • 18 Gaur D, Swaminathan S, Batra JK. Interaction of human pancreatic ribonuclease with human ribonuclease inhibitor: generation of inhibitor-resistant cytotoxic variants. J Biol Chem 2001; 276: 24978-24984.
    CrossrefPubMedSearch in Google Scholar
  • 19 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.
    CrossrefPubMedSearch in Google Scholar
  • 20 Fischer S, Gerriets T, Wessels C. et al. Extracellular RNA mediates endothelial-cell permeability via vascular endothelial growth factor. Blood 2007; 110: 2457-2465.
    CrossrefPubMedSearch in Google Scholar
  • 21 Jaffe EA, Nachman RL, Becker CG. et al. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745-2756.
    CrossrefPubMedSearch in Google Scholar
  • 22 Kanse SM, Benakour O, Kanthou C. et al. Induction of vascular SMC proliferation by urokinase indicates a novel mechanism of action in vasoproliferative disorders. Arterioscler Thromb Vasc Biol 1997; 17: 2848-2854.
    CrossrefPubMedSearch in Google Scholar
  • 23 Rosenthal A, Gotlieb A. Macrovascular endothelial cells from porcine aorta. In: Cell Culture Techniques in Heart and Vessel Research. Springer-Verlag; Heidelberg: 1990. pp. 158-178.
    CrossrefSearch in Google Scholar
  • 24 Weksler BB, Subileau EA Perriere. et al. Blood-brain barrier-specific properties of a human adult brain endothelial cell line. Faseb J 2005; 19: 1872-1874.
    CrossrefPubMedSearch in Google Scholar
  • 25 Edgell C-JS, McDonald CC, Graham JB. Permanent cell line expressing factor VIII-related antigen established by hybridization. Proc Natl Acad Sci USA 1983; 80: 3734-3737.
    CrossrefPubMedSearch in Google Scholar
  • 26 Zimmermann SB, Sandeen G. A sensitive assay for pancreatic ribonuclease. Anal Biochem 1965; 10: 444-449.
    CrossrefPubMedSearch in Google Scholar
  • 27 Blagoveshchenskaya AD, Hannah MJ, Allen S. et al. Selective and Signal-dependent recruitment of membrane proteins to secretory granules formed by heterologously expressed von Willebrand Factor. Mol Biol Cell 2002; 13: 1582-1593.
    CrossrefPubMedSearch in Google Scholar
  • 28 Fischer S, Nishio M, Peters SC. et al. Signaling mechanism of extracellular RNA in endothelial cells. FASEB J 2009; 23: 2100-2109.
    CrossrefPubMedSearch in Google Scholar
  • 29 Reinders JH, De Groot PG, Gonsalves MD. et al. Isolation of a storage and secre-tory organelle containing von Willebrand protein from cultured endothelial cells. Biochim Biophys Acta 1984; 804: 361-369.
    CrossrefPubMedSearch in Google Scholar
  • 30 Wagner DD, Olmsted JB, Marder VJ. Immunolocalization of von Willebrand protein in Weibel-Palade bodies of human endothelial cells. J Cell Biol 1982; 95: 355-360.
    CrossrefPubMedSearch in Google Scholar
  • 31 Fiedler U, Scharpfenecker M, Koidl S. et al. The Tie-2 ligand Angiopoietin-2 is stored in and rapidly released upon stimulation from endothelial cell Weibel-Palade bodies. Blood 2004; 103: 4150-4156.
    CrossrefPubMedSearch in Google Scholar
  • 32 Kaufmann JE, Oksche A, Wollheim CB. et al. Vasopressin-induced von Wille-brand factor secretion from endothelial cells involves V2 receptors and cAMP. J Clin Invest 2000; 106: 107-116.
    CrossrefPubMedSearch in Google Scholar
  • 33 Dardik R, Livnat T, Seligsohn U. Variable effects of alpha v suppression on VEGFR-2 expression in endothelial cells of different vascular beds. Thromb Haemost 2009; 102: 975-982.
    Thieme ConnectPubMedSearch in Google Scholar
  • 34 Sorrentino S, Naddeo M, Russo A. et al. Degradation of double-stranded RNA by huma pancreatic ribonuclease: crucial role of noncatalytic basic amino acid residues. Biochemistry 2003; 42: 10182-10190.
    CrossrefPubMedSearch in Google Scholar
  • 35 Robb AO, Din JN, Mills NL. et al. The influence of the menstrual cycle, normal pregnancy and pre-eclampsia on platelet activation. Thromb Haemost 2010; 103: 372-378.
    Thieme ConnectPubMedSearch in Google Scholar
  • 36 Pavlov N, Hatzi E, Bassaglia Y. et al. Angiogenin distribution in human term placenta and expression by cultured trophoblastic cells. Angiogenesis 2003; 6: 317-330.
    CrossrefPubMedSearch in Google Scholar
  • 37 Rybak SM. Angiogenin mRNA in human tumor and normal cells. Biochem. Biophys Res Commun 1989; 146: 1240-1248.
    PubMedSearch in Google Scholar
  • 38 Loverix S, Steyaert J. Ribonucleases: from prototypes to therapeutic targets?. Curr Med Chem 2003; 10: 779-785.
    CrossrefPubMedSearch in Google Scholar
  • 39 Rajashekhar G, Loganath A, Roy AC. et al. Hypoxia up-regulated angiogenin and down-regulated vascular cell adhesion molecule-1 expression and secretion in human placental trophoblasts. J Soc Gynecol Invest 2005; 12: 310-319.
    CrossrefPubMedSearch in Google Scholar
  • 40 Houge G, Doskeland SO. Divergence towards a dead end? Cleavage of the divergent domains of ribosomal RNA in apoptosis. Experientia 1996; 52: 963-967.
    CrossrefPubMedSearch in Google Scholar
  • 41 Lee FS, Vallee BL. Structure and action of mammalian ribonuclease (angiogenin) inhibitor. Prog Nucl Acid Res Mol Biol 1993; 44: 1-30.
    PubMedSearch in Google Scholar
  • 42 Blackburn P, Moore SA. Pancreatic ribonuclease. In: The Enzymes. 1982. 15 pp. 317-433.
    Search in Google Scholar
  • 43 Wu Y, Mikulski SM, Ardelt W. et al. A cytotoxic ribonuclease. Study of the mechanism of onconase cytotoxicity. J Biol Chem 1993; 268: 10686-10693.
    PubMedSearch in Google Scholar
  • 44 Bonfanti R, Furie BC, Furie B. et al. PADGE (GMP140) is a component of WeibelPalade bodies of human endothelial cells. Blood 1989; 73: 1109-1112.
    PubMedSearch in Google Scholar
  • 45 Vischer UM, Wagner DD. CD63 is a component of Weibel-Palade bodies of human endothelial cells. Blood 1993; 82: 1184-1191.
    PubMedSearch in Google Scholar
  • 46 Huber D, Cramer EM, Kaufmann JE. et al. Tissue-type plasminogen activator (t-PA) is stored in Weibel-Palade bodies in human endothelial cells in vitro and in vivo. Blood 2002; 99: 3637-3645.
    CrossrefPubMedSearch in Google Scholar
  • 47 Muldowney JAS, Painter CA, Sanders-Bush E. et al. Acute tissue-type plasminogen activator release in human microvascular endothelial cells: The roles of Gαq, PLC-β, IP3 and 5,6-epoxyeicosatrienoic acid. Thromb Haemost 2007; 97: 263-271.
    Thieme ConnectPubMedSearch in Google Scholar
  • 48 Wolff JEA, Munstermann G, Grebenkamper K. et al. Gamma-glutamyl-transpeptidase does not act as a cystine transporter in brain microvessels. Neurochem Res 1998; 23: 1175-1178.
    CrossrefPubMedSearch in Google Scholar
  • 49 Hamilton KK, Sims PJ. Changes in cytosolic Ca2+associated with von Willebrand factor release in human endothelial cell monolayers using the fluorescent probe indo-1. J Clin Invest 1987; 79: 600-608.
    CrossrefPubMedSearch in Google Scholar
  • 50 Schluter T, Bohnensack R. Serotonin-induced secretion of von Willebrand factor from human umbilical vein endothelial cells via the cyclic AMP-signaling systems independent of increased cytoplasmic calcium concentration. Biochem Pharmacol 1999; 57: 1191-1197.
    CrossrefPubMedSearch in Google Scholar
  • 51 Bhatia R, Matsushita K, Yamakuchi M. et al. Ceramide triggers Weibel-Palade body exocytosis. Circ Res 2004; 95: 319-324.
    CrossrefPubMedSearch in Google Scholar
  • 52 Löwenberg EC, Charunwatthana P, Cohen S. et al. Severe malaria is associated with a deficiency of von Willebrand factor cleaving protease, ADAMT13. Thromb Haemost 2009; 103: 181-187.
    Thieme ConnectPubMedSearch in Google Scholar
  • 53 Haberichter SS, Merricks EP, Fahs SA. et al. Re-establishment of VWF-dependent Weibel-Palade bodies in VWD endothelial cells. Blood 2005; 105: 145-152.
    CrossrefPubMedSearch in Google Scholar
  • 54 Methia N, Andre P, Denis CV. et al. Localized reduction of atheroscleorosis in von Willebrand factor-deficient mice. Blood 2001; 98: 1424-1428.
    CrossrefPubMedSearch in Google Scholar
  • 55 Barrabes S, Pages-Pons L, Radcliffe CM. et al. Glycosylation of serum ribonuclease 1 indicates a major endothelial origin and reveals an increase in core fucosylation in pancreatic cancer. Glycobiol 2007; 17: 388-400.
    CrossrefPubMedSearch in Google Scholar