High-Dose Epinephrine Enhances Platelet Aggregation at the Expense of Procoagulant Activity

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

doi:10.1055/a-1420-7630

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2021; 121(10): 1337-1344
DOI: 10.1055/a-1420-7630

Cellular Haemostasis and Platelets

High-Dose Epinephrine Enhances Platelet Aggregation at the Expense of Procoagulant Activity

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

› Author Affiliations

Abstract

Platelet activation is characterized by shape change, granule secretion, activation of fibrinogen receptor (glycoprotein IIb/IIIa) sustaining platelet aggregation, and externalization of negatively charged aminophospholipids contributing to platelet procoagulant activity. Epinephrine (EPI) alone is a weak platelet activator. However, it is able to potentiate platelet activation initiated by other agonists. In this work, we investigated the role of EPI in the generation of procoagulant platelets. Human platelets were activated with convulxin (CVX), thrombin (THR) or protease-activated receptor (PAR) agonists, EPI, and combination thereof. Platelet aggregation was assessed by light transmission aggregometry or with PAC-1 binding by flow cytometry. Procoagulant collagen-and-THR (COAT) platelets, induced by combined activation with CVX-and-THR, were visualized by flow cytometry as Annexin-V-positive and PAC-1-negative platelets. Cytosolic calcium fluxes were monitored by flow cytometry using Fluo-3 indicator. EPI increased platelet aggregation induced by all agonist combinations tested. On the other hand, EPI dose-dependently reduced the formation of procoagulant COAT platelets generated by combined CVX-and-THR activation. We observed a decreased Annexin-V-positivity and increased binding of PAC-1 with the triple activation (CVX + THR + EPI) compared with CVX + THR. Calcium mobilization with triple activation was decreased with the higher EPI dose (1,000 µM) compared with CVX + THR calcium kinetics. In conclusion, when platelets are activated with CVX-and-THR, the addition of increasing concentrations of EPI (triple stimulation) modulates platelet response reducing cytosolic calcium mobilization, decreasing procoagulant activity, and enhancing platelet aggregation.

Keywords

platelets - procoagulant platelets - epinephrine - aggregation

Full Text

References

References
1 Agbani EO, Poole AW. Procoagulant platelets: generation, function, and therapeutic targeting in thrombosis. Blood 2017; 130 (20) 2171-2179
2 Reddy EC, Rand ML. Procoagulant phosphatidylserine-exposing platelets in vitro and in vivo . Front Cardiovasc Med 2020; 7: 15
3 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
4 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
5 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
6 Pasalic L, Wing-Lun E, Lau JK. et al. Novel assay demonstrates that coronary artery disease patients have heightened procoagulant platelet response. J Thromb Haemost 2018; 16 (06) 1198-1210
7 Prodan CI, Vincent AS, Dale GL. Coated-platelet levels are elevated in patients with transient ischemic attack. Transl Res 2011; 158 (01) 71-75
8 Prodan CI, Stoner JA, Cowan LD, Dale GL. Higher coated-platelet levels are associated with stroke recurrence following nonlacunar brain infarction. J Cereb Blood Flow Metab 2013; 33 (02) 287-292
9 Prodan CI, Joseph PM, Vincent AS, Dale GL. Coated-platelets in ischemic stroke: differences between lacunar and cortical stroke. J Thromb Haemost 2008; 6 (04) 609-614
10 Prodan CI, Vincent AS, Padmanabhan R, Dale GL. Coated-platelet levels are low in patients with spontaneous intracerebral hemorrhage. Stroke 2009; 40 (07) 2578-2580
11 Prodan CI, Vincent AS, Dale GL. Coated platelet levels correlate with bleed volume in patients with spontaneous intracerebral hemorrhage. Stroke 2010; 41 (06) 1301-1303
12 Prodan CI, Stoner JA, Gordon DL, Dale GL. Cerebral microbleeds in nonlacunar brain infarction are associated with lower coated-platelet levels. J Stroke Cerebrovasc Dis 2014; 23 (05) e325-e330
13 Prodan CI, Vincent AS, Kirkpatrick AC, Hoover SL, Dale GL. Higher levels of coated-platelets are observed in patients with subarachnoid hemorrhage but lower levels are associated with increased mortality at 30 days. J Neurol Sci 2013; 334 (1–2): 126-129
14 Prodan CI, Stoner JA, Dale GL. Lower coated-platelet levels are associated with increased mortality after spontaneous intracerebral hemorrhage. Stroke 2015; 46 (07) 1819-1825
15 Daskalakis M, Colucci G, Keller P. et al. Decreased generation of procoagulant platelets detected by flow cytometric analysis in patients with bleeding diathesis. Cytometry B Clin Cytom 2014; 86 (06) 397-409
16 Dale GL. Procoagulant platelets: further details but many more questions. Arterioscler Thromb Vasc Biol 2017; 37 (09) 1596-1597
17 Norgard NB, Hann CL, Dale GL. Cangrelor attenuates coated-platelet formation. Clin Appl Thromb Hemost 2009; 15 (02) 177-182
18 Norgard NB, Saya S, Hann CL, Hennebry TA, Schechter E, Dale GL. Clopidogrel attenuates coated-platelet production in patients undergoing elective coronary catheterization. J Cardiovasc Pharmacol 2008; 52 (06) 536-539
19 Kotova YN, Ataullakhanov FI, Panteleev MA. Formation of coated platelets is regulated by the dense granule secretion of adenosine 5'diphosphate acting via the P2Y12 receptor. J Thromb Haemost 2008; 6 (09) 1603-1605
20 van der Meijden PE, Feijge MA, Giesen PL, Huijberts M, van Raak LP, Heemskerk JW. Platelet P2Y12 receptors enhance signalling towards procoagulant activity and thrombin generation. A study with healthy subjects and patients at thrombotic risk. Thromb Haemost 2005; 93 (06) 1128-1136
21 Bertaggia Calderara D, Crettaz D, Aliotta A. et al. Generation of procoagulant collagen- and thrombin-activated platelets in platelet concentrates derived from buffy coat: the role of processing, pathogen inactivation, and storage. Transfusion 2018; 58 (10) 2395-2406
22 Dorsam RT, Tuluc M, Kunapuli SP. Role of protease-activated and ADP receptor subtypes in thrombin generation on human platelets. J Thromb Haemost 2004; 2 (05) 804-812
23 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
24 Haaland HD, Holmsen H. Potentiation by adrenaline of agonist-induced responses in normal human platelets in vitro. Platelets 2011; 22 (05) 328-337
25 Lanza F, Beretz A, Stierlé A, Hanau D, Kubina M, Cazenave JP. Epinephrine potentiates human platelet activation but is not an aggregating agent. Am J Physiol 1988; 255 (6, Pt 2): H1276-H1288
26 Ghilotti M, Lova P, Balduini C, Torti M. Epinephrine induces intracellular Ca2+ mobilization in thrombin-desensitized platelets: a role for GPIb-IX-V. Platelets 2007; 18 (02) 135-142
27 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
28 Aliotta A, Bertaggia Calderara D, Zermatten MG, Alberio L. Sodium-calcium exchanger reverse mode sustains dichotomous ion fluxes required for procoagulant COAT platelet formation. Thromb Haemost 2021; 121 (03) 309-321
29 Aliotta A, Bertaggia Calderara D, Alberio L. Flow cytometric monitoring of dynamic cytosolic calcium, sodium, and potassium fluxes following platelet activation. Cytometry A 2020; 97 (09) 933-944
30 von Känel R, Dimsdale JE. Effects of sympathetic activation by adrenergic infusions on hemostasis in vivo. Eur J Haematol 2000; 65 (06) 357-369
31 Austin AW, Wissmann T, von Kanel R. Stress and hemostasis: an update. Semin Thromb Hemost 2013; 39 (08) 902-912
32 Singh S, Malm CJ, Ramström S, Hesse C, Jeppsson A. Adrenaline enhances in vitro platelet activation and aggregation in blood samples from ticagrelor-treated patients. Res Pract Thromb Haemost 2018; 2 (04) 718-725
33 Spalding A, Vaitkevicius H, Dill S, MacKenzie S, Schmaier A, Lockette W. Mechanism of epinephrine-induced platelet aggregation. Hypertension 1998; 31 (02) 603-607
34 Olbrich C, Aepfelbacher M, Siess W. Epinephrine potentiates calcium mobilization and activation of protein kinases in platelets stimulated by ADP through a mechanism unrelated to phospholipase C. Cell Signal 1989; 1 (05) 483-492
35 Singh S, Damén T, Nygren A. et al. Adrenaline improves platelet reactivity in ticagrelor-treated healthy volunteers. Thromb Haemost 2019; 119 (05) 735-743
36 Martin AC, Zlotnik D, Bonete GP. et al. Epinephrine restores platelet functions inhibited by ticagrelor: A mechanistic approach. Eur J Pharmacol 2020; 866: 172798
37 Handtke S, Wesche J, Palankar R, Greinacher A, Thiele T. Function of large and small platelets differs, depending on extracellular calcium availability and type of inductor. Thromb Haemost 2020; 120 (07) 1075-1086
38 van der Meijden PEJ, Heemskerk JWM. Platelet biology and functions: new concepts and clinical perspectives. Nat Rev Cardiol 2019; 16 (03) 166-179
39 Estevez B, Du X. New concepts and mechanisms of platelet activation signaling. Physiology (Bethesda) 2017; 32 (02) 162-177
40 Banga HS, Simons ER, Brass LF, Rittenhouse SE. Activation of phospholipases A and C in human platelets exposed to epinephrine: role of glycoproteins IIb/IIIa and dual role of epinephrine. Proc Natl Acad Sci U S A 1986; 83 (23) 9197-9201
41 Siess W, Weber PC, Lapetina EG. Activation of phospholipase C is dissociated from arachidonate metabolism during platelet shape change induced by thrombin or platelet-activating factor. Epinephrine does not induce phospholipase C activation or platelet shape change. J Biol Chem 1984; 259 (13) 8286-8292
42 Crouch MF, Lapetina EG. A role for Gi in control of thrombin receptor-phospholipase C coupling in human platelets. J Biol Chem 1988; 263 (07) 3363-3371
43 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
44 Andropoulos DB, Soifer SJ, Schreiber MD. Plasma epinephrine concentrations after intraosseous and central venous injection during cardiopulmonary resuscitation in the lamb. J Pediatr 1990; 116 (02) 312-315
45 Wortsman J, Paradis NA, Martin GB. et al. Functional responses to extremely high plasma epinephrine concentrations in cardiac arrest. Crit Care Med 1993; 21 (05) 692-697
46 Fitzcharles-Bowe C, Denkler K, Lalonde D. Finger injection with high-dose (1:1,000) epinephrine: does it cause finger necrosis and should it be treated?. Hand (N Y) 2007; 2 (01) 5-11

Supplementary Material

Supplementary Material