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DOI: 10.1160/TH12-04-0232
Reduction of cAMP and cGMP inhibitory effects in human platelets by MRP4-mediated transport
Financial support: This work was partially supported by a grant from the Italian Ministry of Education, University and Scientific Research (MIUR), partly from PRIN project 2009 to F.M.P. and partly from ex 60%-Ateneo 2008, 2009 to F.M.P.Publication History
Received:
11 April 2012
Accepted after major revision:
09 August 2012
Publication Date:
29 November 2017 (online)
Summary
Cyclic nucleotide-dependent inhibition of platelets represents the most important physiological way to limit thrombus formation. cAMP and cGMP increase in platelets as a consequence of prostacyclin and nitric oxide production by endothelial cells and act through PKA and PKG, respectively. The cytosolic concentration of cyclic nucleotides in platelets is regulated by AC- and GC-dependent synthesis and PDE-dependent degradation. In some cells cyclic nucleotides are eliminated also through MRP4/5/8-dependent efflux. As only MRP4 is expressed in platelets, at high levels in dense granules, we determined its role in the elimination of cyclic nucleotides from platelet cytosol. We studied the effects of MRP4 inhibition on cAMP/cGMP effects in platelets. Cyclic nucleotide inhibitory effects triggered by cAMP and cGMP-elevating agents on platelet aggregation are strongly enhanced by MRP4 inhibition and so is cyclic nucleotide-dependent phosphorylation of the common substrate VASP. MRP4 inhibition decreases cAMP concentration in platelet granules and both cAMP and cGMP compete with an established substrate of MRP4 (fluo-cAMP) for entrance in granules. Here we provide the first evidence of the transport of cyclic nucleotides mediated by MRP4 as part of their physiological mechanism of elimination in human platelets, which might represent a novel target to increase cyclic nucleotide-dependent inhibition.
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References
- 1 Schwarz UR, Walter U, Eigenthaler M. Taming platelets with cyclic nucleotides. Biochem Pharmacol 2001; 62: 1153-1161.
- 2 Walter U, Gambaryan S. cGMP and cGMP-dependent protein kinase in platelets and blood cells. Handb Exp Pharmacol 2009; 191: 533-548.
- 3 Smolenski A. Novel roles of cAMP/cGMP-dependent signaling in platelets. J Thromb Haemost 2012; 10: 167-176.
- 4 Sassi Y, Lipskaia L, Vandecasteele G. et al. Multidrug resistance-associated protein 4 regulates cAMP-dependent signaling pathways and controls human and rat SMC proliferation. J Clin Invest 2008; 118: 2747-2757.
- 5 van Aubel RA, Smeets PH, Peters JG. et al. 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: 595-603.
- 6 Wielinga PR, van der Heijden I, Reid G. et al. Characterization of the MRP4- and MRP5-mediated transport of cyclic nucleotides from intact cells. J Biol Chem 2003; 278: 17664-17671.
- 7 Chen ZS, Lee K, Kruh GD. Transport of cyclic nucleotides and estradiol 17-beta-D-glucuronide by multidrug resistance protein 4. Resistance to 6-mercaptopurine and 6-thioguanine. J Biol Chem 2001; 276: 33747-33754.
- 8 Jedlitschky G, Burchell B, Keppler D. The multidrug resistance protein 5 functions as an ATP-dependent export pump for cyclic nucleotides. J Biol Chem 2000; 275: 30069-30074.
- 9 Guo Y, Kotova E, Chen ZS. et al. MRP8, ATP-binding cassette C11 (ABCC11), is a cyclic nucleotide efflux pump and a resistance factor for fluoropyrimidines 2',3'-dideoxycytidine and 9'-(2'-phosphonylmethoxyethyl)adenine. J Biol Chem 2003; 278: 29509-29514.
- 10 Sager G, Ravna AW. Cellular efflux of cAMP and cGMP - a question about selectivity. Mini Rev Med Chem 2009; 9: 1009-1013.
- 11 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: 3603-3010.
- 12 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: 86-94.
- 13 Fukami MH, Bauer JS, Stewart GJ. et al. An improved method for the isolation of dense storage granules from human platelets. J Cell Biol 1978; 77: 389-399.
- 14 Salganicoff L, Fukami MH. Energy metabolism of blood platelets. I. Isolation and properties of platelet mitochondria. Arch Biochem Biophys 1972; 153: 726-735.
- 15 Reichel V, Masereeuw R, van den Heuvel JJ. et al. Transport of a fluorescent cAMP analog in teleost proximal tubules. Am J Physiol Regul Integr Comp Physiol 2007; 293: R2382-R2389.
- 16 Reichel V, Klas J, Fricker G. et al. Fluo-cAMP is transported by multidrug resistance-associated protein isoform 4 in rat choroid plexus. J Neurochem 2010; 115: 200-208.
- 17 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: 752-761.
- 18 Russel FG, Koenderink JB, Masereeuw R. Multidrug resistance protein 4 (MRP4/ABCC4): a versatile efflux transporter for drugs and signalling molecules. Trends Pharmacol Sci 2008; 29: 200-207.
- 19 Hara Y, Sassi Y, Guibert C. et al. Inhibition of MRP4 prevents and reverses pulmonary hypertension in mice. J Clin Invest 2011; 121: 2888-2897.
- 20 Gradilone A, Pulcinelli FM, Lotti LV. et al. Celecoxib induces MRP-4 in lung cancer cells: therapeutic implications. J Clin Oncol 2007; 25: 4318-4320.
- 21 Gresele P, Momi S, Falcinelli E. Anti-platelet therapy: phosphodiesterase inhibitors. Br J Clin Pharmacol 2011; 72: 634-646.