Superoxide Dismutase 2 is dispensable for platelet functionFinancial support: RS work was funded by F32 HL128008–01 and T32 DK 091317. JR was funded by K01 GM 103806. ASW was funded by R01 HL 126547–01. EB’s work is supported by a Canadian Institutes of Health Research Foundation grant (to EB), and is recipient of a salary award from the Canadian Institutes of Health Research (CIHR). LB is a recipient of a fellowship from The Arthritis Society. This work was supported by NIH Grant U54 HL112311 to ASW and EDA who are both established investigators of the American Heart Association.
12 March 2017
Accepted after major revision: 11 June 2017
08 November 2017 (online)
Increased intracellular reactive oxygen species (ROS) promote platelet activation. The sources of platelet-derived ROS are diverse and whether or not mitochondrial derived ROS, modulates platelet function is incompletely understood. Studies of platelets from patients with sickle cell disease, and diabetes suggest a correlation between mitochondrial ROS and platelet dysfunction. Therefore, we generated mice with a platelet specific knockout of superoxide dismutase 2 (SOD2-KO) to determine if increased mitochondrial ROS increases platelet activation. SOD2-KO platelets demonstrated decreased SOD2 activity and increased mitochondrial ROS, however total platelet ROS was unchanged. Mitochondrial function and content were maintained in non-stimulated platelets. However SOD2-KO platelets demonstrated decreased mitochondrial function following thrombin stimulation. In vitro platelet activation and spreading was normal and in vivo, deletion of SOD2 did not change tail-bleeding or arterial thrombosis indices. In pathophysiological models mediated by platelet-dependent immune mechanisms such as sepsis and autoimmune inflammatory arthritis, SOD2-KO mice were phenotypically identical to wildtype controls. These data demonstrate that increased mitochondrial ROS does not result in platelet dysfunction.
- 1 Handin RI, Karabin R, Boxer GJ. Enhancement of Platelet Function by Superoxide Anion. J Clin Invest 1977; 59: 959-965.
- 2 Salvemini D, de NUcci G, Sneddon JM, Vane JR. Superoxide anions enhance platelet adhesion adn aggregation. Br J Pharmacol 1989; 97: 1145-1150.
- 3 Freedman JE. Oxidative stress and platelets. Arterioscl Thromb Vasc Biol 2008; 28: s11-s16.
- 4 Pignatelli FMP, Lenti L, Gassaniga PP, Violi F. Hydrogen Peroxide Is Involved in Collagen-Induced Platelet Activation. Blood 1998; 91: 484-490.
- 5 Delaney MK, Kim K, Estevez B. et al. Differential Roles of the NADPH-Oxidase 1 and 2 in Platelet Activation and Thrombosis. Arterioscl Thromb Vasc Biol 2016; 36: 846-854.
- 6 Walsh TG, Berndt MC, Carrim N. et al. The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation. Redox Biol 2014; 02: 178-186.
- 7 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.
- 8 Dayal S, Wilson KM, Motto DG. et al. Hydrogen peroxide promotes aging-related platelet hyperactivation and thrombosis. Circulation 2013; 127: 1308-1316.
- 9 Yamagishi S, Edelstein D, Du X, Brownlee M. Hyperglycemia Potentiates Collagen-Induced Platelet Activation Through Mitochondiral Superoxide Overproduction. Diabetes 2001; 50: 1491-1494.
- 10 Karpatkin S. Studies on Human Platelet Glycolysis Effect of Glucose, Cyanide, Insulin, Citrate, and Agglutination and Contraction on Platelet Glycolysis. J Clin Invest 1967; 46: 409-417.
- 11 Sena LA, Chandel NS. Physiological roles of mitochondrial reactive oxygen species. Mol Cell 2012; 48: 158-167.
- 12 Li Y, Huang TT, Carlson EJ. et al. Dilated cardiomyopathy and neonatal lethality in mutant mice lacking manganese superoxide dismutase. Nature Gen 1995; 11: 376-381.
- 13 Van Remmen H, Williams MD, Guo Z. et al. Knockout mice heterozygous for Sod2 show alterations in cardiac mitochondrial function and apoptosis. Am J Physiol Heart Circ Physiol 2001; 281: H1422-H1432.
- 14 Cardenes N, Corey C, Geary L. et al. Platelet bioenergetic screen in sickle cell patients reveals mitochondrial complex V inhibition, which contributes to platelet activation. Blood 2014; 123: 2864-2872.
- 15 Tang WH, Stitham J, Jin Y. et al. Aldose reductase-mediated phosphorylation of p53 leads to mitochondrial dysfunction and damage in diabetic platelets. Circulation 2014; 129: 1598-1609.
- 16 Avila C, Huang RJ, Stevens MV. et al. Platelet mitochondrial dysfunction is evident in type 2 diabetes in association with modifications of mitochondrial anti-oxidant stress proteins. Exp Clin Endocrinol Diabetes 2012; 120: 248-251.
- 17 Wang Z, Cai F, Chen X. et al. The role of mitochondria-derived reactive oxygen species in hyperthermia-induced platelet apoptosis. PloS one 2013; 08: e75044.
- 18 Sjovall F, Morota S, Hansson MJ. et al. Temporal increase of platelet mitochondrial respiration is negatively associated with clinical outcome in patients with sepsis. Crit Care 2010; 14: R214.
- 19 Ikegami T, Suzuki Y, Shimizu T. et al. Model mice for tissue-specific deletion of the manganese superoxide dismutase (MnSOD) gene. Biochem Biophys Res Commun 2002; 296: 729-736.
- 20 Wagner BA, Evig CB, Reszka KJ. et al. Doxorubicin increases intracellular hydrogen peroxide in PC3 prostate cancer cells. Arch Biochem Biophys 2005; 440: 181-190.
- 21 Yusa T, Beckman JS, Crapo JD, Freeman BA. Hyperoxia increases H2O2 production by brain in vivo. J Appl Physiol 1987; 63: 353-358.
- 22 Fink BD, Herlein JA, O’Malley Y. et al. Endothelial cell and platelet bioenergetics: effect of glucose and nutrient composition. PloS one 2012; 07: e39430.
- 23 Schwertz H, Koster S, Kahr WH. et al. Anucleate platelets generate progeny. Blood 2010; 115: 3801-3809.
- 24 Hubbard WJ, Choudhry M, Schwacha MG. et al. Cecal Ligation and Puncture. Shock 2005; 24 (01) 52-57.
- 25 Monach PA, Mathis D, Benoist C. The K/BxN arthritis model. Curr Protoc Immunol 2008; 15 Unit (15) 22.
- 26 Boilard E, Nigrovic PA, Larabee K. et al. Platelets Amplify Inflammation in Arthritis via Collagen-Dependent Microparticle Production. Science 2010; 327: 580-583.
- 27 Adrie JM, Verhoeven MEM, Akkerman JWN. Metabolic Energy is required in Human Platelets at any Stage Durring Optical Aggregation and Secretion. Biochim Biophys Acta 1984; 800: 242-250.
- 28 Xiang B, Zhang G, Guo L. et al. Platelets protect from septic shock by inhibiting macrophage-dependent inflammation via the cyclooxygenase 1 signalling pathway. Nature Commun 2013; 04: 1-12.
- 29 Auchere F, Rusnak F. What is the ultimate fate of superoxide anion in vivo?. J Biol Inorg Chem 2002; 07: 664-667.
- 30 Di Paola M, Lorusso M. Interaction of free fatty acids with mitochondria: coupling, uncoupling and permeability transition. Biochim Biophys Acta 2006; 1757: 1330-1337.
- 31 Christopher J, Barile PCH, Tyvoll David A, Collamn James P, Decreau Rchard A, Brian S. Bull. Inhibiting platelet-stimulated blood coagulation by inhibition of mitochondrial respiration. PNAS 2011; 109: 2539-2543.
- 32 Macarthur H, Westfall TC, Riley DP. et al. Inactivation of catecholamines by superoxide gives new insights on the pathogenesis of septic shock. PNAS 2000; 97: 9753-9758.
- 33 Duttaroy A, Paul A, Kundu M, Belton A. A Sod2 Null Mutation Confers Severely Reduced Adult Life Span in Drosophila. Genetics 2003; 165: 2295-2299.