The acceleration of the propagation phase of thrombin generation in patients with steady-state sickle cell disease is associated with circulating erythrocyte-derived microparticles

 

 Buy Article Permissions and Reprints

Summary

Sickle cell disease (SCD) is linked to hypercoagulability and is characterised by high concentrations of erythrocyte-derived microparticles (Ed-MPs). However, the impact of procoagulant cell-derived microparticles on the thrombin generation process remains unclear. We analysed the alterations of each phase of thrombin generation (TG) in relation to the concentration of erythrocyte- or platelet-derived microparticles (Ed-MPs and Pd-MPs) in a cohort of patients with steady-state SCD. We studied 92 steady-state SCD patients, 19 of which were under treatment with hydroxyurea, and 30 healthy age- and sex-matched individuals. TG was assessed by calibrated automated thrombogram. Ed-MP and Pd-MP expressing or not phosphatidylserine (PS) were determined by means of flow cytometry. Procoagulant phospholipid-dependent activity in the plasma was evaluated by the Procoag-PPL assay. Levels of thrombomodulin and haemoglobin in the plasma as well as red blood cell and reticulocyte counts were measured. SCD patients, independently of the administration of hydroxyurea, were marked by a significant acceleration in the propagation phase of TG which correlated with the Ed-MP/PS⁺ concentration. TG was significantly attenuated in hydroxyurea-treated patients. In conclusion, the acceleration of the propagation phase of TG, driven by Ed-MP/PS⁺, is a major functional alteration in blood coagulation in patients with steady-state SCD. Treatment with hydroxyurea, in addition to the regulation of haemolysis, lowers Ed-MPs and attenuates thrombin generation. The thrombogram could be a useful tool for the diagnosis of hypercoagulability and optimisation of the treatment in patients with SCD.

• References

• 1 Austin H, Key NS, Benson JM. et al. Sickle cell trait and the risk of venous thromboembolism among blacks. Blood 2007; 110: 908-912.
  CrossrefPubMedGoogle Scholar
• 2 Prengler M, Pavlakis SG, Prohovnik I. et al. Sickle cell disease : the neurological complications. Ann Neurol 2002; 51: 543-552.
  CrossrefPubMedGoogle Scholar
• 3 Ataga KI, Cappellini MD, Rachmilewitz EA. Betathalassaemia and sickle cell anaemia as paradigms of hypercoagulability. Br J Haematol 2007; 139: 3-13.
  CrossrefPubMedGoogle Scholar
• 4 Au NH, Wong AY, Vickars L. et al. Two new examples of Hb S. Etienne [beta 92(F8)HisGln] in association with venous thrombosist. Hemoglobin 2009; 33: 95-100.
  CrossrefPubMedGoogle Scholar
• 5 Ataga KI. Hypercoagulability and thrombotic complications in hemolytic anemias. Haematologica 2009; 94: 1481-1484.
  CrossrefPubMedGoogle Scholar
• 6 Hagger D, Wolff S, Owen J. et al. Changes in coagulation and fibrinolysis in patients with sickle cell disease compared with healthy black controls. Blood Coagul Fibrinolysis 1995; 6: 93-99.
  CrossrefPubMedGoogle Scholar
• 7 De Franceschi L, Cappellini MD, Olivieri O. Thrombosis and sickle cell disease. Semin Thromb Hemost 2011; 37: 226-236.
  Article in Thieme ConnectPubMedGoogle Scholar
• 8 Kaul DK, Hebbel RP. Hypoxia/reoxygenation causes inflammatory response in transgenic sickle mice but not in normal mice. J Clin Invest 2000; 106: 411-420.
  CrossrefPubMedGoogle Scholar
• 9 Osarogiagbon UR, Choong S, Belcher JD. et al. Reperfusion injury pathophysiology in sickle transgenic mice. Blood 2000; 96: 314-320.
  PubMedGoogle Scholar
• 10 Gavins FN, Russell J, Senchenkova EL. et al. Mechanisms of enhanced thrombus formation in cerebral microvessels of mice expressing hemoglobin-S. Blood 2011; 117: 4125-4133.
  CrossrefPubMedGoogle Scholar
• 11 Proença-Ferreira R, Franco-Penteado CF, Traina F. et al. Increased adhesive properties of platelets in sickle cell disease: roles for alphaIIb beta3-mediated ligand inding, diminished cAMP signalling and increased phosphodiesterase 3A activity. Br J Haematol 2010; 149: 280-288.
  CrossrefPubMedGoogle Scholar
• 12 van Beers EJ, Schaap MC, Berckmans RJ. et al CURAMA study group. Circulating erythrocyte-derived microparticles are associated with coagulation activation in sickle cell disease. Haematologica 2009; 94: 1513-1519.
  CrossrefPubMedGoogle Scholar
• 13 Allan D, Limbrick AR, Thomas P. et al. Release of spectrin-free spicules on reoxygenation of sickled erythrocytes. Nature 1982; 295: 612-613.
  CrossrefPubMedGoogle Scholar
• 14 Zwaal RFA, Schroit AJ. Pathophysiologic implications of membrane phospholipids asymmetry in blood cells. Blood 1997; 89: 1121-1132.
  PubMedGoogle Scholar
• 15 Setty BN, Kulkani S, Stuart MJ. Role of erythrocyte phosphatidylserine in sickle red cell-endothelial adhesion. Blood 2002; 99: 1564-1571.
  CrossrefPubMedGoogle Scholar
• 16 Tait JF, Gibson D. Measurement of membrane phospholipid asymmetry in normal and sickle-cell erythrocytes by means of annexin V binding. J Lab Clin Med 1994; 123: 741-748.
  PubMedGoogle Scholar
• 17 Stein PD, Beemath A, Meyers FA. et al. Deep venous thrombosis and pulmonary embolism in hospitalized patients with sickle cell disease. Am J Med 2006; 119 897 e7-11.
  PubMedGoogle Scholar
• 18 Hendrick AM. Autoimmune haemolytic anaemia - a high-risk disorder for thromboembolism?. Hematology 2003; 8: 53-56.
  CrossrefPubMedGoogle Scholar
• 19 Devaux PF, Zachowski A. Maintenance and consequences of membrane phospholipid asymmetry. Chem Phys Lipids 1994; 73: 107-120.
  CrossrefPubMedGoogle Scholar
• 20 Fadok VA, Voelker DR, Campbell PA. et al. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J Immunol 1992; 148: 2207-2216.
  PubMedGoogle Scholar
• 21 Hebbel RP, Eaton JW, Balasingam M. et al. Spontaneous oxygen radical generation by sickle erythrocytes. J Clin Invest 1982; 70: 1253-1259.
  CrossrefPubMedGoogle Scholar
• 22 Setty BN, Rao AK, Stuart MJ. Thrombophilia in sickle cell disease: the red cell connection. Blood 2001; 98: 3228-3233.
  CrossrefPubMedGoogle Scholar
• 23 Devaux PF. Static and dynamic lipid asymmetry in cell membranes. Biochemistry 1991; 30: 1163-1173.
  CrossrefPubMedGoogle Scholar
• 24 Westerman MP, Green D, Gilman-Sachs A. et al. Coagulation changes in individual with sickle ceel trait. Am J Hematol 2002; 69: 89-94.
  CrossrefPubMedGoogle Scholar
• 25 Helley D, Girot R, Guillin MC. et al. Sickle cell disease: relation between procoagulant activity of red blood cells from different phenotypes and in vivo blood coagulation activation. Br J Haematol 1997; 99: 268-272.
  CrossrefPubMedGoogle Scholar
• 26 Setty BN, Kulkarni S, Rao AK. et al. Fetal hemoglobin in sickle cell disease: relationship to erythrocyte phosphatidylserine exposure and coagulation activation. Blood 2000; 96: 1119-1124.
  PubMedGoogle Scholar
• 27 Rees DC, Williams TN, Gladwin MT. Sickle-cell disease. Lancet 2010; 376: 2018-2031.
  CrossrefPubMedGoogle Scholar
• 28 Charache S, Terrin ML, Moore RD. et al. Effect of hydroxyurea on the frequency of painful crisis in sickle cell anemia. Investigators of the Multicenter study of Hydroxyurea in sickle cell anemia. N Engl J Med 1995; 332: 1317-1322.
  CrossrefPubMedGoogle Scholar
• 29 Westerman MP, Cole ER, Wu K. The effect of spicules obtained from sickle red cells on clotting activity. Br J Haematol 1984; 56: 557-562.
  CrossrefPubMedGoogle Scholar
• 30 Brummel-Ziedins KE, Orfeo T, Rosendaal FR. et al. Empirical and theoretical phenotypic discrimination. J Thromb Haemost 2009; 7 (Suppl. 01) 181-186.
  CrossrefPubMedGoogle Scholar
• 31 Freyssinet JM, Toti F. Formation of procoagulant microparticles and properties. Thromb Res 2010; 125 (Suppl. 01) S46-48.
  CrossrefPubMedGoogle Scholar
• 32 Piccin A, Murphy WG, Smith OP. Circulating microparticles: pathophysiology and clinical implications. Blood Rev 2007; 21: 157-171.
  CrossrefPubMedGoogle Scholar
• 33 Ataga KI, Key NS. Hypercoagulability in sickle cell disease: new approaches to an old problem. Hematology Am Soc Hematol Educ Prog 2007; 91-96.
  PubMedGoogle Scholar
• 34 Westerman M, Pizzey A, Hirschman J. et al. Microvesicles in haemoglobinopathies offer insights into mechanisms of hypercoagulability, haemolysis and the effects of therapy. Br J Haematol 2008; 142: 126-135.
  CrossrefPubMedGoogle Scholar
• 35 Bridges KR, Barabino GD, Brugnara C. et al. A multiparameter analysis of sickle erythrocytes in patients undergoing hydroxyurea therapy. Blood 1996; 88: 4701-4710.
  PubMedGoogle Scholar
• 36 Covas TC, de Lucenas I, Vianna Palma PBonini. Effects of hydroxyurea on the membrane of erythrocytes and platelets in sickle cell anemia. Haematologica 2004; 89: 273-280.
  PubMedGoogle Scholar
• 37 Setty BN, Kulkarni S, Dampier CD. et al. Fetal haemoglobin in sickle cell anemia: relationship to erythrocyte adhesion markers and adhesion. Blood 2001; 97: 2568-2573.
  CrossrefPubMedGoogle Scholar
• 38 Hebbel RP, Boogaerts MA, Eaton JW. et al. Erythrocyte adherence to endothelium in sickle-cell anemia. A possible determinant of disease severity. N Engl J Med 1980; 302: 992-995.
  CrossrefPubMedGoogle Scholar