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Thromb Haemost 2018; 118(01): 132-142
DOI: 10.1160/TH17-04-0291
Cellular Haemostasis and Platelets
Schattauer GmbH Stuttgart

Release of Platelet-Derived Sphingosine-1-Phosphate Involves Multidrug Resistance Protein 4 (MRP4/ABCC4) and Is Inhibited by Statins

Katja Vogt
,
Shailaja Mahajan-Thakur
,
Robert Wolf
,
Susanne Bröderdorf
,
Conny Vogel
,
Andreas Böhm
,
Christoph A. Ritter
,
Markus Gräler
,
Stefan Oswald
,
Andreas Greinacher
,
Heyo K. Kroemer
,
Gabriele Jedlitschky   *
,
Bernhard H. Rauch   *
Further Information

Publication History

26 April 2017

10 October 2017

Publication Date:
05 January 2018 (online)

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Abstract

Sphingosine-1-phosphate (S1P) is a potent lipid mediator released from activated platelets by an adenosine triphosphate (ATP)-dependent export mechanism. A candidate transport protein is the multidrug resistance protein 4 (MRP4/ABCC4), an ATP-dependent transporter highly expressed in platelets. Furthermore, several statins are known to affect platelet functions and exhibit antithrombotic properties. This study determines the involvement of MRP4 in the transport of S1P and a possible interference by statins. Transport studies in membrane vesicles of Sf9 cells containing recombinant human MRP4 revealed that MRP4 mediates ATP-dependent transport of fluorescein- and tritium-labelled S1P. Also, ATP-dependent S1P transport in platelet membrane vesicles containing endogenous MRP4 was inhibited by the MRP inhibitor MK571 and the MRP4-selective compound Ceefourin-1. Confocal microscopy using fluorescein-labelled S1P as well as boron-dipyrromethene (BODIPY)-labelled sphingosine indicated association of S1P and MRP4 in human platelets. In MRP4-deficient mice, agonist-induced S1P secretion was reduced compared with matched wild-type C57Bl/6 mice and platelet S1P concentrations were lower. Fluvastatin and rosuvastatin interfered with MRP4 function inhibiting ATP-dependent cGMP (cyclic guanosine monophosphate) uptake into MRP4-containing vesicles, inhibited MRP4-mediated S1P transport in vitro and significantly attenuated endogenous S1P release from agonist-activated platelet ex vivo. These data suggest that release of S1P from platelets depends on MRP4 and statins can interfere with this transport process. Potentially, this may be relevant for the pleiotropic anti-inflammatory effects of statins and their effect on modulating atherothrombosis.

Keywords

sphingosine-1-phosphate - MRP4 - platelets - statins - ABC transporter

* Gabriele Jedlitschky and Bernhard H. Rauch contributed equally to this study.


Supplementary Material

 

  • References

  • 1 Nurden AT. Platelets, inflammation and tissue regeneration. Thromb Haemost 2011; 105 (Suppl. 01) S13-S33
    Thieme ConnectPubMedSearch in Google Scholar
  • 2 Obinata H, Hla T. Sphingosine 1-phosphate in coagulation and inflammation. Semin Immunopathol 2012; 34 (01) 73-91
    CrossrefPubMedSearch in Google Scholar
  • 3 Rauch BH. Sphingosine 1-phosphate as a link between blood coagulation and inflammation. Cell Physiol Biochem 2014; 34 (01) 185-196
    CrossrefPubMedSearch in Google Scholar
  • 4 Knapp M, Baranowski M, Czarnowski D. , et al. Plasma sphingosine-1-phosphate concentration is reduced in patients with myocardial infarction. Med Sci Monit 2009; 15 (09) CR490-CR493
    PubMedSearch in Google Scholar
  • 5 Polzin A, Rassaf T, Böhm A. , et al. Aspirin inhibits release of platelet-derived sphingosine-1-phosphate in acute myocardial infarction. Int J Cardiol 2013; 170 (02) e23-e24
    CrossrefPubMedSearch in Google Scholar
  • 6 Jessup W. Lipid metabolism: sources and stability of plasma sphingosine-1-phosphate. Curr Opin Lipidol 2008; 19 (05) 543-544
    CrossrefPubMedSearch in Google Scholar
  • 7 Yatomi Y. Plasma sphingosine 1-phosphate metabolism and analysis. Biochim Biophys Acta 2008; 1780 (03) 606-611
    CrossrefPubMedSearch in Google Scholar
  • 8 Yatomi Y, Ohmori T, Rile G. , et al. Sphingosine 1-phosphate as a major bioactive lysophospholipid that is released from platelets and interacts with endothelial cells. Blood 2000; 96 (10) 3431-3438
    PubMedSearch in Google Scholar
  • 9 English D, Welch Z, Kovala AT. , et al. Sphingosine 1-phosphate released from platelets during clotting accounts for the potent endothelial cell chemotactic activity of blood serum and provides a novel link between hemostasis and angiogenesis. FASEB J 2000; 14 (14) 2255-2265
    CrossrefPubMedSearch in Google Scholar
  • 10 Shimamura K, Takashiro Y, Akiyama N, Hirabayashi T, Murayama T. Expression of adhesion molecules by sphingosine 1-phosphate and histamine in endothelial cells. Eur J Pharmacol 2004; 486 (02) 141-150
    CrossrefPubMedSearch in Google Scholar
  • 11 Xu CB, Hansen-Schwartz J, Edvinsson L. Sphingosine signaling and atherogenesis. Acta Pharmacol Sin 2004; 25 (07) 849-854
    PubMedSearch in Google Scholar
  • 12 Böhm A, Flößer A, Ermler S. , et al. Factor-Xa-induced mitogenesis and migration require sphingosine kinase activity and S1P formation in human vascular smooth muscle cells. Cardiovasc Res 2013; 99 (03) 505-513
    CrossrefPubMedSearch in Google Scholar
  • 13 Olivera A, Allende ML, Proia RL. Shaping the landscape: metabolic regulation of S1P gradients. Biochim Biophys Acta 2013; 1831 (01) 193-202
    CrossrefPubMedSearch in Google Scholar
  • 14 Kobayashi N, Nishi T, Hirata T. , et al. Sphingosine 1-phosphate is released from the cytosol of rat platelets in a carrier-mediated manner. J Lipid Res 2006; 47 (03) 614-621
    CrossrefPubMedSearch in Google Scholar
  • 15 Ulrych T, Böhm A, Polzin A. , et al. Release of sphingosine-1-phosphate from human platelets is dependent on thromboxane formation. J Thromb Haemost 2011; 9 (04) 790-798
    CrossrefPubMedSearch in Google Scholar
  • 16 Mitra P, Oskeritzian CA, Payne SG, Beaven MA, Milstien S, Spiegel S. Role of ABCC1 in export of sphingosine-1-phosphate from mast cells. Proc Natl Acad Sci U S A 2006; 103 (44) 16394-16399
    CrossrefPubMedSearch in Google Scholar
  • 17 Sato K, Malchinkhuu E, Horiuchi Y. , et al. Critical role of ABCA1 transporter in sphingosine 1-phosphate release from astrocytes. J Neurochem 2007; 103 (06) 2610-2619
    PubMedSearch in Google Scholar
  • 18 Hisano Y, Kobayashi N, Yamaguchi A, Nishi T. Mouse SPNS2 functions as a sphingosine-1-phosphate transporter in vascular endothelial cells. PLoS One 2012; 7 (06) e38941
    CrossrefPubMedSearch in Google Scholar
  • 19 Reid G, Wielinga P, Zelcer N. , et al. The human multidrug resistance protein MRP4 functions as a prostaglandin efflux transporter and is inhibited by nonsteroidal antiinflammatory drugs. Proc Natl Acad Sci U S A 2003; 100 (16) 9244-9249
    CrossrefPubMedSearch in Google Scholar
  • 20 Rius M, Thon WF, Keppler D, Nies AT. Prostanoid transport by multidrug resistance protein 4 (MRP4/ABCC4) localized in tissues of the human urogenital tract. J Urol 2005; 174 (06) 2409-2414
    CrossrefPubMedSearch in Google Scholar
  • 21 Rius M, Hummel-Eisenbeiss J, Keppler D. ATP-dependent transport of leukotrienes B4 and C4 by the multidrug resistance protein ABCC4 (MRP4). J Pharmacol Exp Ther 2008; 324 (01) 86-94
    PubMedSearch in Google Scholar
  • 22 Jedlitschky G, Tirschmann K, Lubenow LE. , et al. The nucleotide transporter MRP4 (ABCC4) is highly expressed in human platelets and present in dense granules, indicating a role in mediator storage. Blood 2004; 104 (12) 3603-3610
    CrossrefPubMedSearch in Google Scholar
  • 23 Cheepala SB, Pitre A, Fukuda Y. , et al. The ABCC4 membrane transporter modulates platelet aggregation. Blood 2015; 126 (20) 2307-2319
    CrossrefPubMedSearch in Google Scholar
  • 24 Decouture B, Dreano E, Belleville-Rolland T. , et al. Impaired platelet activation and cAMP homeostasis in MRP4-deficient mice. Blood 2015; 126 (15) 1823-1830
    CrossrefPubMedSearch in Google Scholar
  • 25 Jedlitschky G, Greinacher A, Kroemer HK. Transporters in human platelets: physiologic function and impact for pharmacotherapy. Blood 2012; 119 (15) 3394-3402
    CrossrefPubMedSearch in Google Scholar
  • 26 Hermann A, Rauch BH, Braun M, Schrör K, Weber AA. Platelet CD40 ligand (CD40L)–subcellular localization, regulation of expression, and inhibition by clopidogrel. Platelets 2001; 12 (02) 74-82
    CrossrefPubMedSearch in Google Scholar
  • 27 Keppler D, Jedlitschky G, Leier I. Transport function and substrate specificity of multidrug resistance protein. Methods Enzymol 1998; 292: 607-616
    PubMedSearch in Google Scholar
  • 28 Bode C, Sensken SC, Peest U. , et al. Erythrocytes serve as a reservoir for cellular and extracellular sphingosine 1-phosphate. J Cell Biochem 2010; 109 (06) 1232-1243
    PubMedSearch in Google Scholar
  • 29 Belinsky MG, Guo P, Lee K. , et al. Multidrug resistance protein 4 protects bone marrow, thymus, spleen, and intestine from nucleotide analogue-induced damage. Cancer Res 2007; 67 (01) 262-268
    CrossrefPubMedSearch in Google Scholar
  • 30 Kobayashi N, Kobayashi N, Yamaguchi A, Nishi T. Characterization of the ATP-dependent sphingosine 1-phosphate transporter in rat erythrocytes. J Biol Chem 2009; 284 (32) 21192-21200
    CrossrefPubMedSearch in Google Scholar
  • 31 Jedlitschky G, Leier I, Buchholz U, Center M, Keppler D. ATP-dependent transport of glutathione S-conjugates by the multidrug resistance-associated protein. Cancer Res 1994; 54 (18) 4833-4836
    PubMedSearch in Google Scholar
  • 32 Cheung L, Flemming CL, Watt F. , et al. High-throughput screening identifies Ceefourin 1 and Ceefourin 2 as highly selective inhibitors of multidrug resistance protein 4 (MRP4). Biochem Pharmacol 2014; 91 (01) 97-108
    PubMedSearch in Google Scholar
  • 33 Niessen J, Jedlitschky G, Grube M. , et al. Expression of ABC-type transport proteins in human platelets. Pharmacogenet Genomics 2010; 20 (06) 396-400
    CrossrefPubMedSearch in Google Scholar
  • 34 van Aubel RA, Smeets PH, Peters JG, Bindels RJ, Russel FG. The MRP4/ABCC4 gene encodes a novel apical organic anion transporter in human kidney proximal tubules: putative efflux pump for urinary cAMP and cGMP. J Am Soc Nephrol 2002; 13 (03) 595-603
    PubMedSearch in Google Scholar
  • 35 Knauer MJ, Urquhart BL, Meyer zu Schwabedissen HE. , et al. Human skeletal muscle drug transporters determine local exposure and toxicity of statins. Circ Res 2010; 106 (02) 297-306
    CrossrefPubMedSearch in Google Scholar
  • 36 Kantola T, Backman JT, Niemi M, Kivistö KT, Neuvonen PJ. Effect of fluconazole on plasma fluvastatin and pravastatin concentrations. Eur J Clin Pharmacol 2000; 56 (03) 225-229
    CrossrefPubMedSearch in Google Scholar
  • 37 Niessen J, Jedlitschky G, Grube M. , et al. Human platelets express organic anion-transporting peptide 2B1, an uptake transporter for atorvastatin. Drug Metab Dispos 2009; 37 (05) 1129-1137
    CrossrefPubMedSearch in Google Scholar
  • 38 Mattiello T, Guerriero R, Lotti LV. , et al. Aspirin extrusion from human platelets through multidrug resistance protein-4-mediated transport: evidence of a reduced drug action in patients after coronary artery bypass grafting. J Am Coll Cardiol 2011; 58 (07) 752-761
    CrossrefPubMedSearch in Google Scholar
  • 39 Borgognone A, Pulcinelli FM. Reduction of cAMP and cGMP inhibitory effects in human platelets by MRP4-mediated transport. Thromb Haemost 2012; 108 (05) 955-962
    Thieme ConnectPubMedSearch in Google Scholar
  • 40 Slot AJ, Molinski SV, Cole SP. Mammalian multidrug-resistance proteins (MRPs). Essays Biochem 2011; 50 (01) 179-207
    CrossrefPubMedSearch in Google Scholar
  • 41 Mahajan-Thakur S, Sostmann BD, Fender AC. , et al. Sphingosine-1-phosphate induces thrombin receptor PAR-4 expression to enhance cell migration and COX-2 formation in human monocytes. J Leukoc Biol 2014; 96 (04) 611-618
    CrossrefPubMedSearch in Google Scholar
  • 42 Skottheim IB, Gedde-Dahl A, Hejazifar S, Hoel K, Asberg A. Statin induced myotoxicity: the lactone forms are more potent than the acid forms in human skeletal muscle cells in vitro. Eur J Pharm Sci 2008; 33 (4–5): 317-325
    CrossrefPubMedSearch in Google Scholar
  • 43 Golan K, Vagima Y, Ludin A. , et al. S1P promotes murine progenitor cell egress and mobilization via S1P1-mediated ROS signaling and SDF-1 release. Blood 2012; 119 (11) 2478-2488
    CrossrefPubMedSearch in Google Scholar
  • 44 Bendall LJ, Basnett J. Role of sphingosine 1-phosphate in trafficking and mobilization of hematopoietic stem cells. Curr Opin Hematol 2013; 20 (04) 281-288
    CrossrefPubMedSearch in Google Scholar
  • 45 Xu H, Yang YJ, Yang T, Qian HY. Statins and stem cell modulation. Ageing Res Rev 2013; 12 (01) 1-7
    CrossrefPubMedSearch in Google Scholar
  • 46 Han X, Yang N, Cui Y, Xu Y, Dang G, Song C. Simvastatin mobilizes bone marrow stromal cells migrating to injured areas and promotes functional recovery after spinal cord injury in the rat. Neurosci Lett 2012; 521 (02) 136-141
    CrossrefPubMedSearch in Google Scholar
  • 47 Huu DL, Matsushita T, Jin G. , et al. FTY720 ameliorates murine sclerodermatous chronic graft-versus-host disease by promoting expansion of splenic regulatory cells and inhibiting immune cell infiltration into skin. Arthritis Rheum 2013; 65 (06) 1624-1635
    CrossrefPubMedSearch in Google Scholar