Plasma microparticles of sickle patients during crisis or ...

THROMBOSIS AND HEMOSTASIS| JULY 9, 2020

Plasma microparticles of sickle patients during crisis or taking hydroxyurea modify endothelium inflammatory properties

Yohann Garnier, Séverine Ferdinand, Marie Garnier, Kizzy-Clara Cita, Régine Hierso, Aurélie Claes, Philippe Connes, Marie-Dominique Hardy-Dessources, Claudine Lapouméroulie, Nathalie Lemonne, Maryse Etienne-Julan, Wassim El Nemer, Marc Romana

Blood (2020) 136 (2): 247–256.

https://doi.org/10.1182/blood.2020004853

Key Points

Plasma MPs released during crisis in SS patients increase endothelial ICAM-1 levels and neutrophil adhesion.

HU treatment lowers PS exposure on MPs, which abolishes their proinflammatory properties.

Abstract

Microparticles (MPs) are submicron extracellular vesicles exposing phosphatidylserine (PS), detected at high concentration in the circulation of sickle cell anemia (SS) patients. Several groups studied the biological effects of MPs generated ex vivo. Here, we analyzed for the first time the impact of circulating MPs on endothelial cells (ECs) from 60 sickle cell disease (SCD) patients. MPs were collected from SCD patients and compared with MPs isolated from healthy individuals (AA). Other plasma MPs were purified from SS patients before and 2 years after the onset of hydroxyurea (HU) treatment or during a vaso-occlusive crisis and at steady-state. Compared with AA MPs, SS MPs increased EC ICAM-1 messenger RNA and protein levels, as well as neutrophil adhesion. We showed that ICAM-1 overexpression was primarily caused by MPs derived from erythrocytes, rather than from platelets, and that it was abolished by MP PS capping using annexin V. MPs from SS patients treated with HU were less efficient to induce a proinflammatory phenotype in ECs compared with MPs collected before therapy. In contrast, MPs released during crisis increased ICAM-1 and neutrophil adhesion levels, in a PS-dependent manner, compared with MPs collected at steady-state. Furthermore, neutrophil adhesion was abolished by a blocking anti–ICAM-1 antibody. Our study provides evidence that MPs play a key role in SCD pathophysiology by triggering a proinflammatory phenotype of ECs. We also uncover a new mode of action for HU and identify potential therapeutics: annexin V and anti–ICAM-1 antibodies.

Subjects:

Red Cells, Iron, and Erythropoiesis, Sickle Cell Disease, Thrombosis and Hemostasis

Topics:

adhesions, cell-derived microparticles, endothelium, hydroxyurea, intercellular adhesion molecule 1, massively-parallel genome sequencing, mucopolysaccharidoses, neutrophils, plasma, electroconvulsive therapy

REFERENCES

1.Rees DC, Williams TN, Gladwin MT. Sickle-cell disease. Lancet. 2010;376(9757):2018-2031.

2.Saint-Martin C, Romana M, Bibrac A, et al. Universal newborn screening for haemoglobinopathies in Guadeloupe (French West Indies): a 27-year experience. J Med Screen. 2013;20(4):177-182.

3.Frenette PS. Sickle cell vasoocclusion: heterotypic, multicellular aggregations driven by leukocyte adhesion. Microcirculation. 2004;11(2):167-177.

4.Kaul DK, Hebbel RP. Hypoxia/reoxygenation causes inflammatory response in transgenic sickle mice but not in normal mice. J Clin Invest. 2000;106(3):411-420.

5.Chirico EN, Pialoux V. Role of oxidative stress in the pathogenesis of sickle cell disease. IUBMB Life. 2012;64(1):72-80.

6.Zhang D, Xu C, Manwani D, Frenette PS. Neutrophils, platelets, and inflammatory pathways at the nexus of sickle cell disease pathophysiology. Blood. 2016;127(7):801-809.

7.Su Y, Lei X, Wu L, Liu L. The role of endothelial cell adhesion molecules P-selectin, E-selectin and intercellular adhesion molecule-1 in leucocyte recruitment induced by exogenous methylglyoxal. Immunology. 2012;137(1):65-79.

8.Diamant M, Tushuizen ME, Sturk A, Nieuwland R. Cellular microparticles: new players in the field of vascular disease? Eur J Clin Invest. 2004;34(6):392-401.

9.Shet AS, Aras O, Gupta K, et al. Sickle blood contains tissue factor-positive microparticles derived from endothelial cells and monocytes. Blood. 2003;102(7):2678-2683.

10.Gerotziafas GT, Van Dreden P, Chaari M, et al. The acceleration of the propagation phase of thrombin generation in patients with steady-state sickle cell disease is associated with circulating erythrocyte-derived microparticles. Thromb Haemost. 2012;107(6):1044-1052.

11.Nébor D, Romana M, Santiago R, et al. Fetal hemoglobin and hydroxycarbamide moduate both plasma concentration and cellular origin of circulating microparticles in sickle cell anemia children. Haematologica. 2013;98(6):862-867.

12.Tantawy AA, Adly AA, Ismail EA, Habeeb NM, Farouk A. Circulating platelet and erythrocyte microparticles in young children and adolescents with sickle cell disease: Relation to cardiovascular complications. Platelets. 2013;24(8):605-614.

13.van Tits LJ, van Heerde WL, Landburg PP, et al. Plasma annexin A5 and microparticle phosphatidylserine levels are elevated in sickle cell disease and increase further during painful crisis. Biochem Biophys Res Commun. 2009;390(1):161-164.

14.Hierso R, Lemonne N, Villaescusa R, et al. Exacerbation of oxidative stress during sickle vaso-occlusive crisis is associated with decreased anti-band 3 autoantibodies rate and increased red blood cell-derived microparticle level: a prospective study. Br J Haematol. 2017;176(5):805-813.

15.Olatunya OS, Lanaro C, Longhini AL, et al. Red blood cells microparticles are associated with hemolysis markers and may contribute to clinical events among sickle cell disease patients. Ann Hematol. 2019;98(11):2507-2521.

16.van Beers EJ, Schaap MCL, Berckmans RJ, et al; CURAMA study group. Circulating erythrocyte-derived microparticles are associated with coagulation activation in sickle cell disease. Haematologica. 2009;94(11):1513-1519.

17.Burger D, Schock S, Thompson CS, Montezano AC, Hakim AM, Touyz RM. Microparticles: biomarkers and beyond. Clin Sci (Lond). 2013;124(7):423-441.

18.Ruf W, Rehemtulla A, Morrissey JH, Edgington TS. Phospholipid-independent and -dependent interactions required for tissue factor receptor and cofactor function. J Biol Chem. 1991;266(4):2158-2166.

19.Bach R, Gentry R, Nemerson Y. Factor VII binding to tissue factor in reconstituted phospholipid vesicles: induction of cooperativity by phosphatidylserine. Biochemistry. 1986;25(14):4007-4020.

20.Nebor D, Bowers A, Connes P, et al. Plasma concentration of platelet-derived microparticles is related to painful vaso-occlusive phenotype severity in sickle cell anemia. PLoS One. 2014;9(1):e87243.

21.Marsh A, Schiffelers R, Kuypers F, et al. Microparticles as biomarkers of osteonecrosis of the hip in sickle cell disease. Br J Haematol. 2015;168(1):135-138.

22.Vats R, Brzoska T, Bennewitz MF, et al. Platelet extracellular vesicles drive inflammasome-IL-1β-dependent lung injury in sickle cell disease. Am. J. Respir. Crit. Care Med. 2020;201(1):33-46.

23.Camus SM, Gausserès B, Bonnin P, et al. Erythrocyte microparticles can induce kidney vaso-occlusions in a murine model of sickle cell disease. Blood. 2012;120(25):5050-5058.

24.Bochmann I, Ebstein F, Lehmann A, et al. T lymphocytes export proteasomes by way of microparticles: a possible mechanism for generation of extracellular proteasomes. J Cell Mol Med. 2014;18(1):59-68.

25.Wang JG, Williams JC, Davis BK, et al. Monocytic microparticles activate endothelial cells in an IL-1β-dependent manner. Blood. 2011;118(8):2366-2374.

26.Garnier Y, Ferdinand S, Connes P, et al. Decrease of externalized phosphatidylserine density on red blood cell-derived microparticles in SCA patients treated with hydroxycarbamide. Br J Haematol. 2018;182(3):448-451.

27.Robert S, Poncelet P, Lacroix R, Raoult D, Dignat-George F. More on: calibration for the measurement of microparticles: value of calibrated polystyrene beads for flow cytometry-based sizing of biological microparticles. J Thromb Haemost. 2011;9(8):1676-1678, NaN-1682.

28.Lacroix R, Plawinski L, Robert S, et al. Leukocyte- and endothelial-derived microparticles: a circulating source for fibrinolysis. Haematologica. 2012;97(12):1864-1872.

29.Schweitzer KM, Vicart P, Delouis C, et al. Characterization of a newly established human bone marrow endothelial cell line: distinct adhesive properties for hematopoietic progenitors compared with human umbilical vein endothelial cells. Lab Invest. 1997;76(1):25-36.

30.Brun M, Bourdoulous S, Couraud PO, Elion J, Krishnamoorthy R, Lapoumeroulie C. Hydroxyurea downregulates endothelin-1 gene expression and upregulates ICAM-1 gene expression in cultured human endothelial cells. Pharmacogenomics J. 2003;3(4):215-226.

31.Tone M, Tone Y, Fairchild PJ, Wykes M, Waldmann H. Regulation of CD40 function by its isoforms generated through alternative splicing. Proc Natl Acad Sci USA. 2001;98(4):1751-1756.

32.Turhan A, Weiss LA, Mohandas N, Coller BS, Frenette PS. Primary role for adherent leukocytes in sickle cell vascular occlusion: a new paradigm. Proc Natl Acad Sci USA. 2002;99(5):3047-3051.

33.Setty BNY, Betal SG. Microvascular endothelial cells express a phosphatidylserine receptor: a functionally active receptor for phosphatidylserine-positive erythrocytes. Blood. 2008;111(2):905-914.

34.Rautou PE, Leroyer AS, Ramkhelawon B, et al. Microparticles from human atherosclerotic plaques promote endothelial ICAM-1-dependent monocyte adhesion and transendothelial migration. Circ Res. 2011;108(3):335-343.

35.Al-Nedawi K, Meehan B, Kerbel RS, Allison AC, Rak J. Endothelial expression of autocrine VEGF upon the uptake of tumor-derived microvesicles containing oncogenic EGFR. Proc Natl Acad Sci USA. 2009;106(10):3794-3799.

36.Garnier Y, Ferdinand S, Etienne-Julan M, et al. Differences of microparticle patterns between sickle cell anemia and hemoglobin SC patients. PLoS One. 2017;12(5):e0177397.

37.Barry OP, Praticò D, Savani RC, FitzGerald GA. Modulation of monocyte-endothelial cell interactions by platelet microparticles. J Clin Invest. 1998;102(1):136-144.

38.Nomura S, Tandon NN, Nakamura T, Cone J, Fukuhara S, Kambayashi J. High-shear-stress-induced activation of platelets and microparticles enhances expression of cell adhesion molecules in THP-1 and endothelial cells. Atherosclerosis. 2001;158(2):277-287.

39.Camus SM, De Moraes JA, Bonnin P, et al. Circulating cell membrane microparticles transfer heme to endothelial cells and trigger vasoocclusions in sickle cell disease. Blood. 2015;125(24):3805-3814.

40.Frenette PS, Atweh GF. Sickle cell disease: old discoveries, new concepts, and future promise. J Clin Invest. 2007;117(4):850-858.

41.Belcher JD, Mahaseth H, Welch TE, et al. Critical role of endothelial cell activation in hypoxia-induced vasoocclusion in transgenic sickle mice. Am J Physiol Heart Circ Physiol. 2005;288(6):2715-2725.

42.Ataga KI, Kutlar A, Kanter J, et al. Crizanlizumab for the prevention of pain crises in sickle cell disease. N Engl J Med. 2017;376(5):429-439.

43.Kutlar A, Embury SH. Cellular adhesion and the endothelium: P-selectin. Hematol Oncol Clin North Am. 2014;28(2):323-339.

44.Lard LR, Mul FPJ, de Haas M, Roos D, Duits AJ. Neutrophil activation in sickle cell disease. J Leukoc Biol. 1999;66(3):411-415.

45.Lum AF, Wun T, Staunton D, Simon SI. Inflammatory potential of neutrophils detected in sickle cell disease. Am J Hematol. 2004;76(2):126-133.

46.Canalli AA, Franco-Penteado CF, Saad STO, Conran N, Costa FF. Increased adhesive properties of neutrophils in sickle cell disease may be reversed by pharmacological nitric oxide donation. Haematologica. 2008;93(4):605-609.

47.Hidalgo A, Chang J, Jang JE, Peired AJ, Chiang EY, Frenette PS. Heterotypic interactions enabled by polarized neutrophil microdomains mediate thromboinflammatory injury. Nat Med. 2009;15(4):384-391.

48.Chiang EY, Hidalgo A, Chang J, Frenette PS. Imaging receptor microdomains on leukocyte subsets in live mice. Nat Methods. 2007;4(3):219-222.

49.Tsao PS, Ma XL, Lefer AM. Activated neutrophils aggravate endothelial dysfunction after reperfusion of the ischemic feline myocardium. Am Heart J. 1992;123(6):1464-1471.

50.Jang J-E, Hod EA, Spitalnik SL, Frenette PS. CXCL1 and its receptor, CXCR2, mediate murine sickle cell vaso-occlusion during hemolytic transfusion reactions. J Clin Invest. 2011;121(4):1397-1401.

51.Lang F, Abed M, Lang E, Föller M. Oxidative stress and suicidal erythrocyte death. Antioxid Redox Signal. 2014;21(1):138-153.

52.Barber LA, Palascak MB, Joiner CH, Franco RS. Aminophospholipid translocase and phospholipid scramblase activities in sickle erythrocyte subpopulations. Br J Haematol. 2009;146(4):447-455.

53.Cao W, Huang J, Wu J, et al. Study of 99mTc-annexin V uptake in apoptotic cell models of Parkinson’s disease. Nucl Med Commun. 2007;28(12):895-901.

54.Lahorte CMM, van de Wiele C, Bacher K, et al. Biodistribution and dosimetry study of 123I-rh-annexin V in mice and humans. Nucl Med Commun. 2003;24(8):871-880.

55.Cornelissen B, Lahorte C, Kersemans V, et al. In vivo apoptosis detection with radioiodinated Annexin V in LoVo tumour-bearing mice following tipifarnib (Zarnestra, R115777) farnesyltransferase inhibitor therapy. Nucl Med Biol. 2005;32(3):233-239.

56.Belhocine T, Steinmetz N, Hustinx R, et al. Increased uptake of the apoptosis-imaging agent (99m)Tc recombinant human Annexin V in human tumors after one course of chemotherapy as a predictor of tumor response and patient prognosis. Clin Cancer Res. 2002;8(9):2766-2774.

57.Kartachova MS, Valdés Olmos RA, Haas RL, Hoebers FJ, van Herk M, Verheij M. 99mTc-HYNIC-rh-annexin-V scintigraphy: visual and quantitative evaluation of early treatment-induced apoptosis to predict treatment outcome. Nucl Med Commun. 2008;29(1):39-44.

58.Rybczynska AA, Boersma HH, de Jong S, et al. Avenues to molecular imaging of dying cells: focus on cancer. Med Res Rev. 2018;38(6):1713-1768.

59.Rand ML, Wang H, Pluthero FG, et al. Diannexin, an annexin A5 homodimer, binds phosphatidylserine with high affinity and is a potent inhibitor of platelet-mediated events during thrombus formation. J Thromb Haemost. 2012;10(6):1109-1119.

60.Shen XD, Ke B, Zhai Y, et al. Diannexin, a novel annexin V homodimer, protects rat liver transplants against cold ischemia-reperfusion injury. Am J Transplant. 2007;7(11):2463-2471.

61.Teoh NC, Ito Y, Field J, et al. Diannexin, a novel annexin V homodimer, provides prolonged protection against hepatic ischemia-reperfusion injury in mice. Gastroenterology. 2007;133(2):632-646.

62.Wever KE, Wagener FADTG, Frielink C, et al. Diannexin protects against renal ischemia reperfusion injury and targets phosphatidylserines in ischemic tissue. PLoS One. 2011;6(8):e24276.

63.McVey M, Tabuchi A, Kuebler WM. Microparticles and acute lung injury. Am J Physiol Lung Cell Mol Physiol. 2012;303(5):L364-L381.

64.Wu D, Birukov K. Endothelial cell mechano-metabolomic coupling to disease states in the lung microvasculature. Front Bioeng Biotechnol. 2019;July 19(7):172.

65.Michiels C, Arnould T, Remacle J. Endothelial cell responses to hypoxia: initiation of a cascade of cellular interactions. Biochim Biophys Acta. 2000;1497(1):1-10.

66.Robert S, Lacroix R, Poncelet P, et al. High-sensitivity flow cytometry provides access to standardized measurement of small-size microparticles--brief report. Arterioscler Thromb Vasc Biol. 2012;32(4):1054-1058.

67.Hebbel RP, Key NS. Microparticles in sickle cell anaemia: promise and pitfalls. Br J Haematol. 2016;174(1):16-29.

68.Zennadi R, Whalen EJ, Soderblom EJ, et al. Erythrocyte plasma membrane-bound ERK1/2 activation promotes ICAM-4-mediated sickle red cell adhesion to endothelium. Blood. 2012;119(5):1217-1227.

69.Telen MJ. Beyond hydroxyurea: new and old drugs in the pipeline for sickle cell disease. Blood. 2016;127(7):810-819.

70.Francis RB Jr., Johnson CS. Vascular occlusion in sickle cell disease: current concepts and unanswered questions. Blood. 1991;77(7):1405-1414.

71.Hebbel RP, Vercellotti GM. The endothelial biology of sickle cell disease. J Lab Clin Med. 1997;129(3):288-293.

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