Platelet NOX, a novel target for anti-thrombotic treatment

Summary

There is a growing body of evidence to suggest that reactive oxidant species (ROS) including O₂ ⁻, OH⁻ or H₂O₂ act as second messengers to activate platelets via 1) calcium mobilisation, 2) nitric oxide (NO) inac-tivation, and 3) interaction with arachidonic to give formation of isoprostanes. Among the enzymes generating ROS formation NOX2, the catalytic core of NADPH oxidase (NOX), plays a prominent role as shown by the almost absent ROS production by platelets taken from patients with hereditary deficiency of NOX2. Experimental and clinical studies provided evidence that NOX2 is implicated in platelet activation. Thus, impaired platelet activation has been detected in patients with NOX2 hereditary deficiency. Similarly, normal platelets added with NOX2 specific inhibitors disclosed impaired platelet activation along with ROS down-regulation. Accordingly, animals prone to atherosclerosis treated with apocynin, a NOX inhibitor, showed reduced platelet adhesion and atherosclerotic plaque. Furthermore, a significant association between NOX2 up-regulation and platelet activation has been detected in patients at athero-thrombotic risk, but a cause-effect relationship needs to be established. These findings may represent a rationale to plan interventional trials with NOX inhibitors to establish if blocking NOX2 or other NOX isoforms may represent a novel anti-platelet approach.

Keywords

Platelets - oxidative stress - NOX - reactive oxygen species

• References

• 1 Sugamura K, Keaney Jr JF. Reactive oxygen species in cardiovascular disease. Free Radical Biol Med 2011; 51: 978-992.
• 2 Violi F, Pignatelli P. Platelet oxidative stress and thrombosis. Thromb Res 2012; 129: 378-381.
• 3 Dayal S, Wilson KM, Motto DG. et al. Hydrogen peroxide promotes aging-related platelet hyperactivation and thrombosis. Circulation 2013; 127: 1308-1316.
• 4 Pignatelli P, Carnevale R, Di Santo S. et al. Inherited human gp91phox deficiency is associated with impaired isoprostane formation and platelet dysfunction. Arterioscler Thromb Vasc Biol 2011; 31: 423-434.
• 5 Pignatelli P, Carnevale R, Pastori D. et al. Immediate antioxidant and antiplatelet effect of atorvastatin via inhibition of Nox2. Circulation 2012; 126: 92-103.
• 6 Lambeth JD. NOX enzymes and the biology of reactive oxygen. Nature reviews Immunology 2004; 04: 181-189.
• 7 Babior BM, Lambeth JD, Nauseef W. The neutrophil NADPH oxidase. Arch Biochem Biophys 2002; 397: 342-344.
• 8 Quie PG, White JG, Holmes B. et al. In vitro bactericidal capacity of human polymorphonuclear leukocytes: diminished activity in chronic granulomatous disease of childhood. J Clin Invest 1967; 46: 668-679.
• 9 Dinauer MC, Pierce EA, Bruns GA. et al. Human neutrophil cytochrome b light chain (p22-phox). Gene structure, chromosomal location, and mutations in cytochrome-negative autosomal recessive chronic granulomatous disease. J Clin Invest 1990; 86: 1729-1737.
• 10 Parkos CA, Dinauer MC, Jesaitis AJ. et al. Absence of both the 91kD and 22kD subunits of human neutrophil cytochrome b in two genetic forms of chronic granulomatous disease. Blood 1989; 73: 1416-1420.
• 11 Stasia MJ, Bordigoni P, Martel C. et al. A novel and unusual case of chronic granulomatous disease in a child with a homozygous 36-bp deletion in the CYBA gene (A22 (0)) leading to the activation of a cryptic splice site in intron 4. Human Genetics 2002; 110: 444-450.
• 12 Groemping Y, Lapouge K, Smerdon SJ. et al. Molecular basis of phosphorylation-induced activation of the NADPH oxidase. Cell 2003; 113: 343-355.
• 13 Sumimoto H, Hata K, Mizuki K. et al. Assembly and activation of the phagocyte NADPH oxidase. Specific interaction of the N-terminal Src homology 3 domain of p47phox with p22phox is required for activation of the NADPH oxidase. J Biol Chem 1996; 271: 22152-22158.
• 14 Han CH, Freeman JL, Lee T. et al. Regulation of the neutrophil respiratory burst oxidase. Identification of an activation domain in p67 (phox). J Biol Chem 1998; 273: 16663-16668.
• 15 Diebold BA, Bokoch GM. Molecular basis for Rac2 regulation of phagocyte NADPH oxidase. Nature Immunol 2001; 02: 211-215.
• 16 Koga H, Terasawa H, Nunoi H. et al. Tetratricopeptide repeat (TPR) motifs of p67 (phox) participate in interaction with the small GTPase Rac and activation of the phagocyte NADPH oxidase. J Biol Chem 1999; 274: 25051-25060.
• 17 Li JM, Shah AM. Intracellular localisation and preassembly of the NADPH oxidase complex in cultured endothelial cells. J Biol Chem 2002; 277: 19952-19960.
• 18 Heymes C, Bendall JK, Ratajczak P. et al. Increased myocardial NADPH oxidase activity in human heart failure. J Am Coll Cardiol 2003; 41: 2164-2171.
• 19 Piccoli C, Ria R, Scrima R. et al. Characterisation of mitochondrial and extramitochondrial oxygen consuming reactions in human hematopoietic stem cells. Novel evidence of the occurrence of NAD (P)H oxidase activity. J Biol Chem 2005; 280: 26467-26476.
• 20 Pignatelli P, Sanguigni V, Lenti L. et al. gp91phox-dependent expression of platelet CD40 ligand. Circulation 2004; 110: 1326-1329.
• 21 Banfi B, Clark RA, Steger K. et al. Two novel proteins activate superoxide generation by the NADPH oxidase NOX1. J Biol Chem 2003; 278: 3510-3513.
• 22 Suh YA, Arnold RS, Lassegue B. et al. Cell transformation by the superoxidegenerating oxidase Mox1. Nature 1999; 401: 79-82.
• 23 Lassegue B, Sorescu D, Szocs K. et al. Novel gp91 (phox) homologues in vascular smooth muscle cells : nox1 mediates angiotensin II-induced superoxide formation and redox-sensitive signalling pathways. Circulation Res 2001; 88: 888-894.
• 24 Kobayashi S, Nojima Y, Shibuya M. et al. Nox1 regulates apoptosis and potentially stimulates branching morphogenesis in sinusoidal endothelial cells. Exp Cell Res 2004; 300: 455-462.
• 25 Ambasta RK, Kumar P, Griendling KK. et al. Direct interaction of the novel Nox proteins with p22phox is required for the formation of a functionally active NADPH oxidase. J Biol Chem 2004; 279: 45935-45941.
• 26 Kawahara T, Ritsick D, Cheng G. et al. Point mutations in the proline-rich region of p22phox are dominant inhibitors of Nox1- and Nox2-dependent reactive oxygen generation. J Biol Chem 2005; 280: 31859-31869.
• 27 Kawahara T, Kohjima M, Kuwano Y. et al. Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells. Am J Physiol Cell Physiol 2005; 288: C450-457.
• 28 Hu T, Ramachandrarao SP, Siva S. et al. Reactive oxygen species production via NADPH oxidase mediates TGF-beta-induced cytoskeletal alterations in endothelial cells. Am J Physiol Renal Physiol 2005; 289: F816-825.
• 29 Ellmark SH, Dusting GJ, Fui MN. et al. The contribution of Nox4 to NADPH oxidase activity in mouse vascular smooth muscle. Cardiovasc Res 2005; 65: 495-504.
• 30 Colston JT, de la Rosa SD, Strader JR. et al. H2O2 activates Nox4 through PLA2-dependent arachidonic acid production in adult cardiac fibroblasts. FEBS Lett 2005; 579: 2533-2540.
• 31 Geiszt M, Kopp JB, Varnai P. et al. Identification of renox, an NAD (P)H oxidase in kidney. Proc Natl Acad Sci USA 2000; 97: 8010-8014.
• 32 Shiose A, Kuroda J, Tsuruya K. et al. A novel superoxide-producing NAD (P)H oxidase in kidney. J Biol Chem 2001; 276: 1417-1423.
• 33 Martyn KD, Frederick LM, von Loehneysen K. et al. Functional analysis of Nox4 reveals unique characteristics compared to other NADPH oxidases. Cell Signal 2006; 18: 69-82.
• 34 Takac I, Schroder K, Zhang L. et al. The E-loop is involved in hydrogen peroxide formation by the NADPH oxidase Nox4. J Biol Chem 2011; 286: 13304-13313.
• 35 Cheng G, Cao Z, Xu X. et al. Homologs of gp91phox: cloning and tissue expression of Nox3, Nox4, and Nox5. Gene 2001; 269: 131-140.
• 36 Banfi B, Molnar G, Maturana A. et al. A Ca (2+)-activated NADPH oxidase in testis, spleen, and lymph nodes. J Biol Chem 2001; 276: 37594-37601.
• 37 Banfi B, Tirone F, Durussel I. et al. Mechanism of Ca2+ activation of the NADPH oxidase 5 (NOX5). J Biol Chem 2004; 279: 18583-18591.
• 38 Seno T, Inoue N, Gao D. et al. Involvement of NADH/NADPH oxidase in human platelet ROS production. Thromb Res 2001; 103: 399-409.
• 39 Krotz F, Sohn HY, Gloe T. et al. NAD (P)H oxidase-dependent platelet superoxide anion release increases platelet recruitment. Blood 2002; 100: 917-924.
• 40 Krotz F, Sohn HY, Pohl U. Reactive oxygen species: players in the platelet game. Arterioscler Thromb Vasc Biol 2004; 24: 1988-1996.
• 41 Cardoso AR, Chausse B, da Cunha FM. et al. Mitochondrial compartmentalisation of redox processes. Free Radical Biol Med 2012; 52: 2201-2208.
• 42 Leo R, Pratico D, Iuliano L. et al. Platelet activation by superoxide anion and hydroxyl radicals intrinsically generated by platelets that had undergone anoxia and then reoxygenated. Circulation 1997; 95: 885-891.
• 43 Pignatelli P, Pulcinelli FM, Lenti L. et al. Hydrogen peroxide is involved in collagen-induced platelet activation. Blood 1998; 91: 484-490.
• 44 Cadenas E. Basic mechanisms of antioxidant activity. BioFactors 1997; 06: 391-397.
• 45 Jin RC, Mahoney CE, Coleman Anderson L. et al. Glutathione peroxidase-3 deficiency promotes platelet-dependent thrombosis in vivo. Circulation 2011; 123: 1963-1973.
• 46 Brigelius-Flohe R, Maiorino M. Glutathione peroxidases. Biochim Biophys Acta 2013; 1830: 3289-3303.
• 47 Freedman JE, Loscalzo J, Benoit SE. et al. Decreased platelet inhibition by nitric oxide in two brothers with a history of arterial thrombosis. J Clin Invest 1996; 97: 979-987.
• 48 Dangel O, Mergia E, Karlisch K. et al. Nitric oxide-sensitive guanylyl cyclase is the only nitric oxide receptor mediating platelet inhibition. J Thromb Haemost 2010; 08: 1343-1352.
• 49 Ignarro LJ. Nitric oxide as a unique signalling molecule in the vascular system: a historical overview. J Physiol Pharmacol 2002; 53: 503-514.
• 50 Naseem KM, Riba R. Unresolved roles of platelet nitric oxide synthase. J Thromb Haemost 2008; 06: 10-19.
• 51 Friebe A, Mergia E, Dangel O. et al. Fatal gastrointestinal obstruction and hypertension in mice lacking nitric oxide-sensitive guanylyl cyclase. Proc Natl Acad Sci USA 2007; 104: 7699-7704.
• 52 Morrow JD. Quantification of isoprostanes as indices of oxidant stress and the risk of atherosclerosis in humans. Arterioscler Thromb Vasc Biol 2005; 25: 279-286.
• 53 FitzGerald GA. Mechanisms of platelet activation: thromboxane A2 as an amplifying signal for other agonists. Am J Cardiol 1991; 68: 11B-15B.
• 54 Morrow JD, Hill KE, Burk RF. et al. A series of prostaglandin F2-like compounds are produced in vivo in humans by a non-cyclooxygenase, free radical-catalysed mechanism. Proc Natl Acad Sci USA 1990; 87: 9383-9387.
• 55 Pratico D, Lawson JA, FitzGerald GA. Cyclooxygenase-dependent formation of the isoprostane, 8-epi prostaglandin F2 alpha. J Biol Chem 1995; 270: 9800-9808.
• 56 Smith CC, Betteridge DJ, Nourooz-Zadeh J. The generation of the F (2)-isoprostane 8-epi-PGF (2alpha) by human platelets on collagen stimulation. Platelets 1999; 10: 253-256.
• 57 Klein T, Reutter F, Schweer H. et al. Generation of the isoprostane 8-epi-prostaglandin F2alpha in vitro and in vivo via the cyclooxygenases. J Pharmacol Exp Therap 1997; 282: 1658-1665.
• 58 Freedman JE. Oxidative stress and platelets. Arterioscler Thromb Vasc Biol 2008; 28: s11-16.
• 59 Pignatelli P, Carnevale R, Di Santo S. et al. Rosuvastatin reduces platelet recruitment by inhibiting NADPH oxidase activation. Biochem Pharmacol 2012; 84: 1635-1642.
• 60 Dobrucki LW, Kalinowski L, Dobrucki IT. et al. Statin-stimulated nitric oxide release from endothelium. Med Sci Monitor 2001; 07: 622-627.
• 61 Pignatelli P, Di Santo S, Buchetti B. et al. Polyphenols enhance platelet nitric oxide by inhibiting protein kinase C-dependent NADPH oxidase activation: effect on platelet recruitment. FASEB J 2006; 20: 1082-1089.
• 62 Pignatelli P, Ghiselli A, Buchetti B. et al. Polyphenols synergistically inhibit oxidative stress in subjects given red and white wine. Atherosclerosis 2006; 188: 77-83.
• 63 Carnevale R, Loffredo L, Pignatelli P. et al. Dark chocolate inhibits platelet isoprostanes via NOX2 down-regulation in smokers. J Thromb Haemost 2012; 10: 125-132.
• 64 Corti R, Flammer AJ, Hollenberg NK. et al. Cocoa and cardiovascular health. Circulation 2009; 119: 1433-1441.
• 65 Qin F, Simeone M, Patel R. Inhibition of NADPH oxidase reduces myocardial oxidative stress and apoptosis and improves cardiac function in heart failure after myocardial infarction. Free Radical Biol Med 2007; 43: 271-281.
• 66 Stef G, Csiszar A, Ziangmin Z. et al. Inhibition of NAD (P)H oxidase attenuates aggregation of platelets from high-risk cardiac patients with aspirin resistance. Pharmacol Reports 2007; 59: 428-436.
• 67 Begonja AJ, Gambaryan S, Geiger J. et al. Platelet NAD (P)H-oxidase-generated ROS production regulates alphaIIbbeta3-integrin activation independent of the NO/cGMP pathway. Blood 2005; 106: 2757-2760.
• 68 Pignatelli P, Tanzilli G, Carnevale R. et al. Ascorbic acid infusion blunts CD40L upregulation in patients undergoing coronary stent. Cardiovasc Therap 2011; 29: 385-394.
• 69 Basili S, Pignatelli P, Tanzilli G. et al. Anoxia-reoxygenation enhances platelet thromboxane A2 production via reactive oxygen species-generated NOX2: effect in patients undergoing elective percutaneous coronary intervention. Arterioscler Thromb Vasc Biol 2011; 31: 1766-1771.
• 70 Cangemi R, Pignatelli P, Carnevale R. et al. Platelet isoprostane overproduction in diabetic patients treated with aspirin. Diabetes 2012; 61: 1626-1632.
• 71 Ferreiro JL, Gomez-Hospital JA, Angiolillo DJ. Platelet abnormalities in diabetes mellitus. Diabetes Vasc Dis Res 2010; 07: 251-259.
• 72 Davi G, Guagnano MT, Ciabattoni G. et al. Platelet activation in obese women: role of inflammation and oxidant stress. J Am Med Assoc 2002; 288: 2008-2014.
• 73 Angelico F, Loffredo L, Pignatelli P. et al. Weight loss is associated with improved endothelial dysfunction via NOX2-generated oxidative stress downregulation in patients with the metabolic syndrome. Int Emerg Med 2012; 07: 219-227.
• 74 Loffredo L, Martino F, Carnevale R. et al. Obesity and hypercholesterolemia are associated with NOX2 generated oxidative stress and arterial dysfunction. J Pediat 2012; 161: 1004-1009.
• 75 Davi G, Chiarelli F, Santilli F. et al. Enhanced lipid peroxidation and platelet activation in the early phase of type 1 diabetes mellitus: role of interleukin-6 and disease duration. Circulation 2003; 107: 3199-3203.
• 76 Davi G, Ciabattoni G, Consoli A. et al. In vivo formation of 8-iso-prostaglandin f2alpha and platelet activation in diabetes mellitus: effects of improved metabolic control and vitamin E supplementation. Circulation 1999; 99: 224-229.
• 77 Carnevale R, Iuliano L, Nocella C. et al. Relationship Between Platelet and Urinary 8-Iso-PGF2alpha Levels in Subjects With Different Degrees of NOX2 Regulation. J Am Heart Assoc 2013; 02: e000198.
• 78 Violi F, Pignatelli P, Pignata C. et al. Reduced atherosclerotic burden in subjects with genetically determined low oxidative stress. Arterioscler Thromb Vasc Biol 2013; 33: 406-412.
• 79 Loffredo L, Carnevale R, Sanguigni V. et al. Does NADPH Oxidase Deficiency Cause Artery Dilatation in Humans?. Antiox Redox Signal 2013; 18: 1491-1496.