Increased platelet aggregation and in vivo platelet activation after granulocyte colony-stimulating factor administration

Alexander O. Spiel; Johann Bartko; Michael Schwameis; Christa Firbas; Jolanta Siller-Matula; Matthias Schuetz; Manuela Weigl; Bernd Jilma

doi:10.1160/TH10-08-0530

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 105(04): 655-662
DOI: 10.1160/TH10-08-0530

Platelets and Blood Cells

Schattauer GmbH

Increased platelet aggregation and in vivo platelet activation after granulocyte colony-stimulating factor administration

A randomised controlled trial

Authors

Alexander O. Spiel

¹Department of Clinical Pharmacology, Medical University of Vienna, Vienna, Austria

²Department of Emergency Medicine, Medical University of Vienna, Vienna, Austria
Johann Bartko

¹Department of Clinical Pharmacology, Medical University of Vienna, Vienna, Austria
Michael Schwameis

¹Department of Clinical Pharmacology, Medical University of Vienna, Vienna, Austria
Christa Firbas

¹Department of Clinical Pharmacology, Medical University of Vienna, Vienna, Austria
Jolanta Siller-Matula

¹Department of Clinical Pharmacology, Medical University of Vienna, Vienna, Austria
Matthias Schuetz

³Department of Nuclear Medicine, Medical University of Vienna, Vienna, Austria
Manuela Weigl

⁴Department of Anaesthesiology, General Intensive Care and Pain Control, Medical University of Vienna, Vienna, Austria
Bernd Jilma

¹Department of Clinical Pharmacology, Medical University of Vienna, Vienna, Austria

Abstract

Summary

Granulocyte colony-stimulating factor (G-CSF) stimulates the bone marrow to produce granulocytes and stem cells and is widely used to accelerate neutrophil recovery after chemotherapy. Interestingly, specific G-CSF receptors have been demonstrated not only on myeloid cells, but also on platelets. Data on the effects of G-CSF on platelet function are limited and partly conflicting. The objective of this study was to determine the effect of G-CSF on platelet aggregation and in vivo platelet activation. Seventy-eight, healthy volunteers were enrolled into this randomised, placebo-controlled trial. Subjects received 5 μg/kg methionyl human granulocyte colony-stimulating factor (r-metHuG-CSF, filgrastim) or placebo subcutaneously for four days. We determined platelet aggregation with a whole blood impedance aggregometer with various, clinically relevant platelet agonists (adenosine diphosphate [ADP], collagen, arachidonic acid [AA], ristocetin and thrombin receptor activating peptide 6 [TRAP]). Filgrastim injection significantly enhanced ADP (+40%), collagen (+60%) and AA (+75%) -induced platelet aggregation (all p<0.01 as compared to placebo and p<0.001 as compared to baseline). In addition, G-CSF enhanced ristocetin-induced platelet aggregation (+18%) whereas TRAP-induced platelet aggregation decreased slightly (-14%) in response to filgrastim. While baseline aggregation with all agonists was only slightly but insignificantly higher in women than in men, this sex difference was enhanced by G-CSF treatment, and became most pronounced for ADP after five days (p<0.001). Enhanced platelet aggregation translated into a 75% increase in platelet activation as measured by circulating soluble P-selectin. G-CSF enhances platelet aggregation and activation in humans. This may put patients suffering from cardiovascular disease and cancer at risk for thrombotic events.

Keywords

G-CSF - platelets - whole blood aggregometry - randomised controlled trial - gender difference

Full Text

References

References
1 Lieschke GJ, Burgess AW. Granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor (1). N Engl J Med 1992; 327: 28-35.
2 Majolino I, Cavallaro AM, Scime R. Peripheral blood stem cells for allogeneic transplantation. Bone Marrow Transplant 1996; 18 (Suppl. 02) 171-174.
3 Metcalf D. The molecular biology and functions of the granulocyte-macrophage colony-stimulating factors. Blood 1986; 67: 257-267.
4 Shimoda K, Okamura S, Harada N. et al. Identification of a functional receptor for granulocyte colony-stimulating factor on platelets. J Clin Invest 1993; 91: 1310-1313.
5 Avenarius HJ, Freund M, Deinhardt J. et al. Effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on circulating platelets. Ann Hematol 1992; 65: 6-9.
6 Kaptan K, Ifran A, Beyan C. et al. Recombinant human granulocyte colony-stimulating factor (rhG-CSF) promotes in vitro platelet aggregation. Hematology 2007; 12: 441-444.
7 Shimoda K, Okamura S, Inaba S. et al. Granulocyte colony-stimulating factor and platelet aggregation. Lancet 1993; 341: 633.
8 LeBlanc R, Roy J, Demers C. et al. A prospective study of G-CSF effects on hemostasis in allogeneic blood stem cell donors. Bone Marrow Transplant 1999; 23: 991-996.
9 Anderlini P, Champlin RE. Biologic and molecular effects of granulocyte colony-stimulating factor in healthy individuals: recent findings and current challenges. Blood 2008; 111: 1767-1772.
10 Spiel AO, Siller-Matula J, Firbas C. et al. Single dose granulocyte colony-stimulating factor markedly enhances shear-dependent platelet function in humans. Platelets 2010; 21: 464-469.
11 Chong BH, Murray B, Berndt MC. et al. Plasma P-selectin is increased in thrombotic consumptive platelet disorders. Blood 1994; 83: 1535-1541.
12 Jilma B, Dirnberger E, Eichler HG. et al. Sex differences in circulating P-selectin, E-selectin and thrombomodulin. Br J Haematol 1996; 95: 575-576.
13 Stohlawetz P, Hergovich N, Stiegler G. et al. Differential induction of P-selectin expression on platelets by two cell separators during plateletpheresis and the effect of gender on the release of soluble P-selectin. Transfusion 1998; 38: 24-30.
14 Mayr FB, Spiel AO, Leitner JM. et al. Duffy antigen modifies the chemokine response in human endotoxemia. Crit Care Med 2008; 36: 159-165.
15 Patriccia R, Antonucci E, Maggini N. et al. Comparison of methods for monitoring residual platelet reactivity after clopidogrel by point-of-care tests on whole blood in high-risk patients. Thromb Haemost 2010; 104: 287-292.
16 Derhaschnig U, Jilma B. Assessment of platelets and the endothelium in patients presenting with acute coronary syndromes--is there a future?. Thromb Haemost 2009; 102: 1144-1148.
17 Siller-Matula JM, Gouya G, Wolzt M. et al. Cross validation of the Multiple Electrode Aggregometry. A prospective trial in healthy volunteers. Thromb Haemost 2009; 102: 397-403.
18 Sibbing D, Schulz S, Braun S. et al. Antiplatelet effects of clopidogrel and bleeding in patients undergoing coronary stent placement. J Thromb Haemost 2010; 8: 250-256.
19 Kalb ML, Potura L, Scharbert G. et al. The effect of ex vivo anticoagulants on whole blood platelet aggregation. Platelets 2009; 20: 7-11.
20 Siller-Matula JM, Christ G, Lang IM. et al. Multiple electrode aggregometry predicts stent thrombosis better than the vasodilator-stimulated phosphoprotein phosphorylation assay. J Thromb Haemost 2010; 8: 351-359.
21 Sibbing D, Braun S, Jawansky S. et al. Assessment of ADP-induced platelet aggregation with light transmission aggregometry and multiple electrode platelet aggregometry before and after clopidogrel treatment. Thromb Haemost 2008; 99: 121-126.
22 Sibbing D, Morath T, Braun S. et al. Clopidogrel response status assessed with Multiplate point-of-care analyses and the incidence and timing of stent thrombosis over six months following coronary stenting. Thromb Haemost 2010; 103: 151-159.
23 Kotzailias N, Graninger M, Knechtelsdorfer M. et al. Acute effects of hyperglycaemia on plasma concentration of soluble P-selectin and von Willebrand factor in healthy volunteers -a prospective randomised double blind controlled study. Thromb Res 2009; 123: 452-459.
24 Derhaschnig U, Reiter R, Knobl P. et al. Recombinant human activated protein C (rhAPC; drotrecogin alfa [activated]) has minimal effect on markers of coagulation, fibrinolysis, and inflammation in acute human endotoxemia. Blood 2003; 102: 2093-2098.
25 Stohlawetz P, Horvath M, Pernerstorfer T. et al. Effects of nitric oxide on platelet activation during plateletpheresis and in vivo tracking of biotinylated platelets in humans. Transfusion 1999; 39: 506-514.
26 Blann AD, Nadar SK, Lip GY. The adhesion molecule P-selectin and cardiovascular disease. Eur Heart J 2003; 24: 2166-2179.
27 Mayr FB, Spiel AO, Leitner JM. et al. Racial differences in endotoxin-induced tissue factor-triggered coagulation. J Thromb Haemost 2009; 7: 634-640.
28 Anderlini P, Przepiorka D, Seong D. et al. Clinical toxicity and laboratory effects of granulocyte-colony-stimulating factor (filgrastim) mobilization and blood stem cell apheresis from normal donors, and analysis of charges for the procedures. Transfusion 1996; 36: 590-595.
29 Jilma B, Hergovich N, Stohlawetz P. et al. Circadian variation of granulocyte colony stimulating factor levels in man. Br J Haematol 1999; 106: 368-370.
30 Stohlawetz PJ, Dzirlo L, Hergovich N. et al. Effects of erythropoietin on platelet reactivity and thrombopoiesis in humans. Blood 2000; 95: 2983-2989.
31 Haiden N, Cardona F, Schwindt J. et al. Changes in thrombopoiesis and platelet reactivity in extremely low birth weight infants undergoing erythropoietin therapy for treatment of anaemia of prematurity. Thromb Haemost 2005; 93: 118-123.
32 Falanga A, Marchetti M, Evangelista V. et al. Neutrophil activation and hemostatic changes in healthy donors receiving granulocyte colony-stimulating factor. Blood 1999; 93: 2506-2514.
33 van Os R, van Schie ML, Willemze R. et al. Proteolytic enzyme levels are increased during granulocyte colony-stimulating factor-induced hematopoietic stem cell mobilization in human donors but do not predict the number of mobilized stem cells. J Hematother Stem Cell Res 2002; 11: 513-521.
34 Cerletti C, Evangelista V, Molino M. et al. Platelet activation by polymorphonu-clear leukocytes: role of cathepsin G and P-selectin. Thromb Haemost 1995; 74: 218-223.
35 Renesto P, Chignard M. Enhancement of cathepsin G-induced platelet activation by leukocyte elastase: consequence for the neutrophil-mediated platelet activation. Blood 1993; 82: 139-144.
36 Wohner N, Keresztes Z, Sotonyi P. et al. Neutrophil granulocyte-dependent proteolysis enhances platelet adhesion to the arterial wall under high-shear flow. J Thromb Haemost 2010; 8: 1624-1631.
37 Willis F, Theti D, Dean S. et al. Pegfilgrastim successfully mobilizes megakaryocyte progenitors into the peripheral blood in subjects with solid tumours. Bone Marrow Transplant 2008; 42: 167-173.
38 Lee DS, Larson MG, Lunetta KL. et al. Clinical and genetic correlates of soluble P-selectin in the community. J Thromb Haemost 2008; 6: 20-31.
39 Jilma B, Hildebrandt J, Kapiotis S. et al. Effects of estradiol on circulating P-selectin. J Clin Endocrinol Metab 1996; 81: 2350-2355.
40 Ridker PM, Buring JE, Rifai N. Soluble P-selectin and the risk of future cardiovascular events. Circulation 2001; 103: 491-495.
41 Kisucka J, Chauhan AK, Zhao BQ. et al. Elevated levels of soluble P-selectin in mice alter blood-brain barrier function, exacerbate stroke, and promote atherosclerosis. Blood 2009; 113: 6015-6022.
42 Elwood PC, Beswick AD, Sharp DS. et al. Whole blood impedance platelet aggregometry and ischemic heart disease. The Caerphilly Collaborative Heart Disease Study. Arteriosclerosis 1990; 10: 1032-1036.
43 Elwood PC, Renaud S, Sharp DS. et al. Ischemic heart disease and platelet aggregation. The Caerphilly Collaborative Heart Disease Study. Circulation 1991; 83: 38-44.
44 Fukumoto Y, Miyamoto T, Okamura T. et al. Angina pectoris occurring during granulocyte colony-stimulating factor-combined preparatory regimen for auto-logous peripheral blood stem cell transplantation in a patient with acute myelogenous leukaemia. Br J Haematol 1997; 97: 666-668.
45 Vij R, Adkins DR, Brown RA. et al. Unstable angina in a peripheral blood stem and progenitor cell donor given granulocyte-colony-stimulating factor. Transfusion 1999; 39: 542-543.
46 Hill JM, Syed MA, Arai AE. et al. Outcomes and risks of granulocyte colony-stimulating factor in patients with coronary artery disease. J Am Coll Cardiol 2005; 46: 1643-1648.
47 Zohlnhofer D, Ott I, Mehilli J. et al. Stem cell mobilization by granulocyte colony-stimulating factor in patients with acute myocardial infarction: a randomized controlled trial. J Am Med Assoc 2006; 295: 1003-1010.