Sodium–Calcium Exchanger Reverse Mode Sustains Dichotomous Ion Fluxes Required for Procoagulant COAT Platelet Formation

Alessandro Aliotta; Debora Bertaggia Calderara; Maxime G. Zermatten; Lorenzo Alberio

doi:10.1055/s-0040-171670

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2021; 121(03): 309-321
DOI: 10.1055/s-0040-171670

Cellular Haemostasis and Platelets

Sodium–Calcium Exchanger Reverse Mode Sustains Dichotomous Ion Fluxes Required for Procoagulant COAT Platelet Formation

Autoren

Alessandro Aliotta

¹Hemostasis and Platelet Research Laboratory, Division of Hematology and Central Hematology Laboratory, Lausanne University Hospital (CHUV) and University of Lausanne (UNIL), Lausanne, Switzerland
Debora Bertaggia Calderara

¹Hemostasis and Platelet Research Laboratory, Division of Hematology and Central Hematology Laboratory, Lausanne University Hospital (CHUV) and University of Lausanne (UNIL), Lausanne, Switzerland
Maxime G. Zermatten

¹Hemostasis and Platelet Research Laboratory, Division of Hematology and Central Hematology Laboratory, Lausanne University Hospital (CHUV) and University of Lausanne (UNIL), Lausanne, Switzerland
Lorenzo Alberio

¹Hemostasis and Platelet Research Laboratory, Division of Hematology and Central Hematology Laboratory, Lausanne University Hospital (CHUV) and University of Lausanne (UNIL), Lausanne, Switzerland

Abstract

Procoagulant collagen-and-thrombin (COAT)-activated platelets represent a subpopulation of activated platelets, which retain a coat of prohemostatic proteins and express phosphatidylserine on their surface. Dichotomous intracellular signaling generating procoagulant platelet activity instead of traditional aggregating endpoints is still not fully elucidated. It has been demonstrated that secondary messengers such as calcium and sodium play a critical role in platelet activation. Therefore, we developed a flow cytometric analysis to investigate intracellular ion fluxes simultaneously during generation of aggregating and procoagulant platelets. Human platelets were activated by convulxin-plus-thrombin. Cytosolic calcium, sodium, and potassium ion fluxes were visualized by specific ion probes and analyzed by flow cytometry. We observed high and prolonged intracellular calcium concentration, transient sodium increase, and fast potassium efflux in COAT platelets, whereas aggregating non-COAT platelets rapidly decreased their calcium content, maintaining higher cytosolic sodium, and experiencing lower and slower potassium depletion. Considering these antithetical patterns, we investigated the role of the sodium–calcium exchanger (NCX) during convulxin-plus-thrombin activation. NCX inhibitors, CBDMB and ORM-10103, dose-dependently reduced the global calcium mobilization induced by convulxin-plus-thrombin activation and dose-dependently prevented formation of procoagulant COAT platelets. Our data demonstrate that both NCX modes are used after convulxin-plus-thrombin-induced platelet activation. Non-COAT platelets use forward-mode NCX, thus pumping calcium out and moving sodium in, while COAT platelets rely on reverse NCX function, which pumps additional calcium into the cytosol, by extruding sodium. In conclusion, we described for the first time the critical and dichotomous role of NCX function during convulxin-plus-thrombin-induced platelet activation.

Keywords

blood platelets - procoagulant platelets - ion kinetics - flow cytometry

Volltext

Referenzen

References
1 Clemetson KJ. Platelets and primary haemostasis. Thromb Res 2012; 129 (03) 220-224
2 Heemskerk JW, Mattheij NJ, Cosemans JM. Platelet-based coagulation: different populations, different functions. J Thromb Haemost 2013; 11 (01) 2-16
3 van der Meijden PEJ, Heemskerk JWM. Platelet biology and functions: new concepts and clinical perspectives. Nat Rev Cardiol 2019; 16 (03) 166-179
4 Agbani EO, Poole AW. Procoagulant platelets: generation, function, and therapeutic targeting in thrombosis. Blood 2017; 130 (20) 2171-2179
5 Reddy EC, Rand ML. Procoagulant phosphatidylserine-exposing platelets in vitro and in vivo . Front Cardiovasc Med 2020; 7: 15
6 Alberio L, Safa O, Clemetson KJ, Esmon CT, Dale GL. Surface expression and functional characterization of alpha-granule factor V in human platelets: effects of ionophore A23187, thrombin, collagen, and convulxin. Blood 2000; 95 (05) 1694-1702
7 Alberio L, Ravanat C, Hechler B, Mangin PH, Lanza F, Gachet C. Delayed-onset of procoagulant signalling revealed by kinetic analysis of COAT platelet formation. Thromb Haemost 2017; 117 (06) 1101-1114
8 Dale GL. Procoagulant platelets: further details but many more questions. Arterioscler Thromb Vasc Biol 2017; 37 (09) 1596-1597
9 Dale GL, Friese P, Batar P. et al. Stimulated platelets use serotonin to enhance their retention of procoagulant proteins on the cell surface. Nature 2002; 415 (6868): 175-179
10 Mattheij NJ, Gilio K, van Kruchten R. et al. Dual mechanism of integrin αIIbβ3 closure in procoagulant platelets. J Biol Chem 2013; 288 (19) 13325-13336
11 Mattheij NJ, Swieringa F, Mastenbroek TG. et al. Coated platelets function in platelet-dependent fibrin formation via integrin αIIbβ3 and transglutaminase factor XIII. Haematologica 2016; 101 (04) 427-436
12 Topalov NN, Yakimenko AO, Canault M. et al. Two types of procoagulant platelets are formed upon physiological activation and are controlled by integrin α(IIb)β(3). Arterioscler Thromb Vasc Biol 2012; 32 (10) 2475-2483
13 Mammadova-Bach E, Nagy M, Heemskerk JWM, Nieswandt B, Braun A. Store-operated calcium entry in thrombosis and thrombo-inflammation. Cell Calcium 2019; 77: 39-48
14 Yang H, Kim A, David T. et al. TMEM16F forms a Ca2+-activated cation channel required for lipid scrambling in platelets during blood coagulation. Cell 2012; 151 (01) 111-122
15 Colucci G, Stutz M, Rochat S. et al. The effect of desmopressin on platelet function: a selective enhancement of procoagulant COAT platelets in patients with primary platelet function defects. Blood 2014; 123 (12) 1905-1916
16 Agbani EO, van den Bosch MT, Brown E. et al. Coordinated membrane ballooning and procoagulant spreading in human platelets. Circulation 2015; 132 (15) 1414-1424
17 Harper MT, Londoño JE, Quick K. et al. Transient receptor potential channels function as a coincidence signal detector mediating phosphatidylserine exposure. Sci Signal 2013; 6 (281) ra50
18 Kerbiriou-Nabias D, Arachiche A, Dachary-Prigent J. Phosphatidylserine exposure and calcium-activated potassium efflux in platelets. Br J Haematol 2011; 155 (02) 268-270
19 Mahaut-Smith MP. A role for platelet TRPC channels in the Ca2+ response that induces procoagulant activity. Sci Signal 2013; 6 (281) pe23
20 Aliotta A, Bertaggia Calderara D, Alberio L. Flow cytometric monitoring of dynamic cytosolic calcium, sodium, and potassium fluxes following platelet activation. Cytometry A 2020;
21 Wolfs JL, Wielders SJ, Comfurius P. et al. Reversible inhibition of the platelet procoagulant response through manipulation of the Gardos channel. Blood 2006; 108 (07) 2223-2228
22 Khananshvili D. Sodium-calcium exchangers (NCX): molecular hallmarks underlying the tissue-specific and systemic functions. Pflugers Arch 2014; 466 (01) 43-60
23 Roberts DE, Matsuda T, Bose R. Molecular and functional characterization of the human platelet Na(+) /Ca(2+) exchangers. Br J Pharmacol 2012; 165 (04) 922-936
24 Polgár J, Clemetson JM, Kehrel BE. et al. Platelet activation and signal transduction by convulxin, a C-type lectin from Crotalus durissus terrificus (tropical rattlesnake) venom via the p62/GPVI collagen receptor. J Biol Chem 1997; 272 (21) 13576-13583
25 Topalov NN, Kotova YN, Vasil'ev SA, Panteleev MA. Identification of signal transduction pathways involved in the formation of platelet subpopulations upon activation. Br J Haematol 2012; 157 (01) 105-115
26 Heemskerk JW, Siljander PR, Bevers EM, Farndale RW, Lindhout T. Receptors and signalling mechanisms in the procoagulant response of platelets. Platelets 2000; 11 (06) 301-306
27 do Céu Monteiro M, Sansonetty F, Gonçalves MJ, O'Connor JE. Flow cytometric kinetic assay of calcium mobilization in whole blood platelets using Fluo-3 and CD41. Cytometry 1999; 35 (04) 302-310
28 Jardin I, Ben Amor N, Bartegi A, Pariente JA, Salido GM, Rosado JA. Differential involvement of thrombin receptors in Ca2+ release from two different intracellular stores in human platelets. Biochem J 2007; 401 (01) 167-174
29 Arachiche A, Kerbiriou-Nabias D, Garcin I, Letellier T, Dachary-Prigent J. Rapid procoagulant phosphatidylserine exposure relies on high cytosolic calcium rather than on mitochondrial depolarization. Arterioscler Thromb Vasc Biol 2009; 29 (11) 1883-1889
30 Vergara C, Latorre R, Marrion NV, Adelman JP. Calcium-activated potassium channels. Curr Opin Neurobiol 1998; 8 (03) 321-329
31 Warny M, Kelly CP. Monocytic cell necrosis is mediated by potassium depletion and caspase-like proteases. Am J Physiol 1999; 276 (03) C717-C724
32 Lang PA, Kaiser S, Myssina S, Wieder T, Lang F, Huber SM. Role of Ca2+-activated K+ channels in human erythrocyte apoptosis. Am J Physiol Cell Physiol 2003; 285 (06) C1553-C1560
33 El Kebir D, József L, Khreiss T, Filep JG. Inhibition of K+ efflux prevents mitochondrial dysfunction, and suppresses caspase-3-, apoptosis-inducing factor-, and endonuclease G-mediated constitutive apoptosis in human neutrophils. Cell Signal 2006; 18 (12) 2302-2313
34 Borin M, Siffert W. Stimulation by thrombin increases the cytosolic free Na+ concentration in human platelets. Studies with the novel fluorescent cytosolic Na+ indicator sodium-binding benzofuran isophthalate. J Biol Chem 1990; 265 (32) 19543-19550
35 Sage SO, Rink TJ, Mahaut-Smith MP. Resting and ADP-evoked changes in cytosolic free sodium concentration in human platelets loaded with the indicator SBFI. J Physiol 1991; 441: 559-573
36 Tomasiak M, Stelmach H, Rusak T, Ciborowski M, Radziwon P. The involvement of Na+/K(+)-ATPase in the development of platelet procoagulant response. Acta Biochim Pol 2007; 54 (03) 625-639
37 Roberts DE, Bose R. Reverse mode Na+/Ca2+ exchange in the collagen activation of human platelets. Ann N Y Acad Sci 2002; 976: 345-349
38 Roberts DE, McNicol A, Bose R. Mechanism of collagen activation in human platelets. J Biol Chem 2004; 279 (19) 19421-19430
39 Secondo A, Pannaccione A, Molinaro P. et al. Molecular pharmacology of the amiloride analog 3-amino-6-chloro-5-[(4-chloro-benzyl)amino]-n-[[(2,4-dimethylbenzyl)-amino]iminomethyl]-pyrazinecarboxamide (CB-DMB) as a pan inhibitor of the Na+-Ca2+ exchanger isoforms NCX1, NCX2, and NCX3 in stably transfected cells. J Pharmacol Exp Ther 2009; 331 (01) 212-221
40 Jost N, Nagy N, Corici C. et al. ORM-10103, a novel specific inhibitor of the Na+/Ca2+ exchanger, decreases early and delayed afterdepolarizations in the canine heart. Br J Pharmacol 2013; 170 (04) 768-778
41 Kholmukhamedov A, Janecke R, Choo HJ, Jobe SM. The mitochondrial calcium uniporter regulates procoagulant platelet formation. J Thromb Haemost 2018; 16 (11) 2315-2321
42 Harper MT, Molkentin JD, Poole AW. Protein kinase C alpha enhances sodium-calcium exchange during store-operated calcium entry in mouse platelets. Cell Calcium 2010; 48 (06) 333-340
43 Staiano RI, Granata F, Secondo A. et al. Expression and function of Na+/Ca2+ exchangers 1 and 3 in human macrophages and monocytes. Eur J Immunol 2009; 39 (05) 1405-1418
44 Harper AG, Sage SO. A key role for reverse Na+/Ca2+ exchange influenced by the actin cytoskeleton in store-operated Ca2+ entry in human platelets: evidence against the de novo conformational coupling hypothesis. Cell Calcium 2007; 42 (06) 606-617
45 Lytton J. Na+/Ca2+ exchangers: three mammalian gene families control Ca2+ transport. Biochem J 2007; 406 (03) 365-382
46 Lemos VS, Poburko D, Liao CH, Cole WC, van Breemen C. Na+ entry via TRPC6 causes Ca2+ entry via NCX reversal in ATP stimulated smooth muscle cells. Biochem Biophys Res Commun 2007; 352 (01) 130-134
47 Bye AP, Unsworth AJ, Gibbins JM. Platelet signaling: a complex interplay between inhibitory and activatory networks. J Thromb Haemost 2016; 14 (05) 918-930
48 Heemskerk JW, Harper MT, Cosemans JM, Poole AW. Unravelling the different functions of protein kinase C isoforms in platelets. FEBS Lett 2011; 585 (12) 1711-1716
49 Gilio K, Harper MT, Cosemans JM. et al. Functional divergence of platelet protein kinase C (PKC) isoforms in thrombus formation on collagen. J Biol Chem 2010; 285 (30) 23410-23419
50 Choo HJ, Saafir TB, Mkumba L, Wagner MB, Jobe SM. Mitochondrial calcium and reactive oxygen species regulate agonist-initiated platelet phosphatidylserine exposure. Arterioscler Thromb Vasc Biol 2012; 32 (12) 2946-2955

Zusatzmaterial

Supplementary Material (PDF) (opens in new window)