Different expression of proteins in platelets from aspirin-resistant and aspirin-sensitive patients

Petra J. Mateos-Cáceres; Carlos Macaya; Luis Azcona; Javier Modrego; Estela Mahillo; Esther Bernardo; Antonio Fernández-Ortiz; Antonio J. López-Farré

doi:10.1160/TH09-05-0290

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2010; 103(01): 160-170
DOI: 10.1160/TH09-05-0290

Platelets and Blood Cells

Schattauer GmbH

Different expression of proteins in platelets from aspirin-resistant and aspirin-sensitive patients

Authors

Petra J. Mateos-Cáceres

¹Cardiovascular Research Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain
Carlos Macaya

¹Cardiovascular Research Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain

²Coronary Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain
Luis Azcona

¹Cardiovascular Research Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain
Javier Modrego

¹Cardiovascular Research Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain
Estela Mahillo

¹Cardiovascular Research Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain
Esther Bernardo

²Coronary Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain
Antonio Fernández-Ortiz

²Coronary Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain
Antonio J. López-Farré

¹Cardiovascular Research Unit, Cardiology Department and Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain

Abstract

Summary

The aim of the present study was to analyse differences in the protein expression profile between platelets from aspirin (ASA)-resistant patients and ASA-sensitive patients. We analysed platelets from 51 clinically stable coronary ischaemic patients taking ASA (100 mg/day) divided into ASA-resistant (n=25) and ASA-sensitive (n=26) based on a platelet functionality test (PFA-100). Proteins associated with cytoskeleton, energetic metabolism, oxidative stress, inflammation and cell survival were analysed by two-dimensional electrophoresis and mass spectrometry. The expression of two gelsolin precursor isotypes and one F-acting capping protein isotype was decreased in ASA-resistant platelets (p<0.05). The expression of glyceraldehyde 3-phosphate dehydrogenase was increased in the ASA-resistant platelets (1751.1 ± 220.6 vs. 4273.3 ± 971.7, 95% confidence interval [CI] 1815.11 to 4061.2, p=0.001). It was accompanied by a reduced expression and activity of 1,6-bisphosphate aldolase in platelets without changes in the content of pyruvate. A reduced expression of gluthathione-S-transferase and the protein disulfide isomerase isotype 1 was found in ASA-resistant platelets. The protein expression of the chloride intracellular channel isotype 1 was increased in ASA-resistant platelets (21.3 ± 3.8 vs. 48.8 ± 6.0, CI 29.5 to 45.95, p=0.03) while the expression of two HSP60 and two HSP71 isotypes was decreased. No changes were observed in proteins associated with inflammation. In conclusion, ASA-resistant and ASA-sensitive platelets are different in terms of the level of expression of proteins associated with mechanisms such as energetic metabolism, cytoskeleton, oxidative stress and cell survival which may be associated with their different ability to respond to ASA.

Keywords

Platelet - aspirin resistance - coronary ischaemia - proteins

Full Text

References

References
1 Patrono C. Low-dose aspirin for the prevention of atherothrombosis. N Engl J Med 2005; 353: 2373-2383.
2 Mason PJ, Jacobs AK, Freedman JE. Aspirin resistance and atherothrombotic disease. J Am Col Cardiol 2005; 46: 986-993.
3 Eikelboom JW, Hirsh J, Weitz JI. et al. Aspirin-resistant thromboxane biosynthesis and the risk of myocardial infarction, stroke, or cardiovascular death in patients at high risk for cardiovascular events. Circulation 2002; 105: 1620-1622.
4 Krasopoulos G, Brister SJ, Fraser R. et al. Aspirin resistance and risk of cardiovascular morbidity: systematic review and meta-analysis. Br Med J 2008; 336: 195-198.
5 Crescente M, Di Castelnuovo A, Iacoviello L. et al. Response variability to aspirin as assessed by the platelet function analyzer (PFA)-100. A systematic review. Thromb Haemost 2008; 99: 14-26.
6 Reny JL, De Moerloose P, Dauzat M. et al. Use of the PFA-100 closure time to predict cardiovascular events in aspirin-treated cardiovascular patients: a systematic review and meta-analysis. J Thromb Haemost 2008; 06: 444-450.
7 Kawasaki T, Ozeki Y, Igawa T. et al. Increased platelet sensitivity to collagen in individuals resistant to low-dose aspirin. Stroke 2000; 31: 591-595.
8 Angiolillo DJ, Fernandez-Ortiz A, Bernardo E. et al. Influence of aspirin resistance on platelet function profiles in patients on long-term aspirin and clopidogrel after percutaneous coronary intervention. Am J Cardiol 2006; 97: 38-43.
9 Macchi L, Sorel N, Christiaens L. Aspirin resistance: definitions, mechanisms, prevalence, and clinical significance. Curr Pharm Des 2006; 12: 251-258.
10 Zimmermann N, Hohlfeld T. Clinical implications of aspirin resistance. Thromb Haemost 2008; 100: 379-390.
11 Freedman JE. The aspirin resistance controversy: clinical entity or platelet heterogeneity?. Circulation 2006; 113: 2888-2896.
12 Alonso-Orgaz S, Moreno L, Macaya C. et al. Proteomic study of plasma from moderate hypercholesterolemic patients. J Proteome Res 2006; 05: 2301-2308.
13 Sacristán D, López-Farré AJ, Zamorano-León JJ. et al. Effects of coronary prestenting platelet inhibition on coronary poststenting inflammation. J Cardiovasc Pharmacol 2008; 51: 286-292.
14 López-Farré AJ, Mateos-Cáceres PJ, Sacristán D. et al. Relationship between vitamin D binding protein and aspirin resistance in coronary ischemic patients: a proteomic study. J Proteome Res 2007; 06: 2481-2487.
15 Gum PA, Kottke-Marchant K, Poggio ED. et al. Profile and prevalence of aspirin resistance in patients with cardiovascular disease. Am J Cardiol 2001; 88: 230-235.
16 López-Farré A, Caramelo C, Esteban A. et al. Effects of aspirin on platelet-neutro-phil interactions. Role of nitric oxide and endothelin-1. Circulation 1995; 91: 2080-2088.
17 Mateos-Cáceres PJ, García-Méndez A, López Farré A. et al. Proteomic analysis of plasma from patients during an acute coronary syndrome. J Am Coll Cardiol 2004; 44: 1578-1583.
18 Sacristán D, Marques M, Zamorano-León JJ. et al. Modifications by Olmesartan medoxomil treatment of the platelet protein profile of moderate hypertensive patients. Proteomics Clin Appl 2008; 02: 1300-1312.
19 Misset O, Opperdoes FR. Simultaneous purification of hexokinase, class-I fructose-bisphosphate aldolase, triosephosphate isomerase and phosphoglycerate kinase from Trypanosoma brucei . Eur J Biochem 1984; 144: 475-483.
20 Marques M, Sacristán D, Mateos-Cáceres PJ. et al. Different protein expression in normal and dysfunctional platelets from uremic patients. J Nephrol. 2009 in press.
21 Cambria-Kiely JA, Gandhi PJ. Aspirin resistance and genetic polymorphisms. J Thromb Thrombolysis 2002; 14: 51-58.
22 Hillarp A, Palmqvist B, Lethagen S. et al. Mutations within the cyclooxygenase-1 gene in aspirin non-responders with recurrence of stroke. Thromb Res 2003; 112: 275-283.
23 McKee SA, Sane DC, Deliargyris EN. Aspirin resistance in cardiovascular disease: A review of prevalence, mechanisms and clinical significance. Thromb Haemost 2002; 88: 711-715.
24 Barkalow K, Witke W, Kwiatkowski DJ. et al. Coordinated regulation of platelet actin filament barbed ends by gelsolin and capping protein. J Cell Biol 1996; 134: 389-399.
25 Sun HQ, Yamamoto M, Mejillano M, Yin HL. Gelsolin, a multifunctional actin regulatory protein. J Biol Chem 1999; 274: 33179-33182.
26 Spinardi L, Witke W. Gelsolin and diseases. Subcell Biochem 2007; 45: 55-69.
27 Akerman JWN, Verhoeven AJM. Platelet responses and metabolism. Vol II CRC Press; Boca Raton FL: 1987. pp. 189-213.
28 Sirover MA. New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochimica Biophysica Acta 1999; 1432: 159-184.
29 Schmitz HD, Bereiter-Hahn J. Glyceraldehyde-3-phosphate dehydrogenase associates with actin filaments in serum deprived NIH3T3 cells only. Cell Biol Int 2002; 26: 155-164.
30 Gallogly MM, Mieyal JJ. Mechanisms of reversible protein glutathionylation in redox signaling and oxidative stress. Current Opinion in Pharmacology 2007; 07: 381-391.
31 McDonagh B, Sheehan D. Effect of oxidative stress on protein thiols in the blue mussel Mytilus edulis: Proteomic identification of target proteins. Proteomics 2007; 07: 3395-3403.
32 Mosialou E, Ekström G, Adang AE. et al. Evidence that rat liver microsomal glut-athione transferase is responsible for glutathione-dependent protection against lipid peroxidation. Biochem Pharmacol 1993; 45: 1645-1651.
33 Qi YX, Qu HJ. Rho-GDP dissociation inhibitor alpha downregulated by low shear stress promotes vascular smooth muscle cell migration and apoptosis: a proteomic analysis. Cardiovas Res 2008; 80: 114-122.
34 Mackenzie AB, MacKenzie AB, Mahaut-Smith MP. Chloride channels in excised membrane patches from human platelets: effect of intracellular calcium. Biochim Biophys Acta 1996; 1278: 131-136.
35 Mahaut-Smith MP. Chloride channels in human platelets: evidence for activation by internal calcium. J Membr Biol 1990; 118: 69-75.
36 Poch E, Fernández-Llama P, Botey A. et al. Parathyroid hormone and platelet cytosolic calcium concentration in essential hypertension. Nephrol Dial Transplant 1995; 10: 366-371.
37 Beere HM. Death versus survival: functional interaction between the apoptotic and stress-inducible heat shock protein pathways. J Clin Invest 2005; 115: 2633-2639.
38 Polla BS, Kantengwa S, Francois D. et al. Mitochondria are selective targets for the protective effects of heat shock against oxidative injury. Proc Natl Acad Sci USA 1996; 93: 6458-6463.
39 Polanowska-Grabowska R, Gear AR. Heat-shock proteins and platelet function. Platelets 2000; 01: 6-22.
40 Xanthoudakis S, Roy S, Rasper D. et al. Hsp60 accelerates the maturation of procaspase-3 by upstream activator proteases during apoptosis. EMBO J 1999; 18: 2049-2056.
41 Lindemann S, Tolley ND, Dixon DA. et al. Activated platelets mediate inflammatory signalling by regulated interleukin 1beta synthesis. J Cell Biol 2001; 154: 485-490.
42 Chen WH, Lee PY, Ng W. et al. Aspirin resistance is associated with a high incidence of myonecrosis after non-urgent percutaneous coronary intervention despite clopidogrel pre-treatment. J Am Coll Cardiol 2004; 43: 1122-1126.
43 Prabhakaran S, Wells KR, Lee VH. et al. Prevalence and risk factors for aspirin and clopidogrel resistance in cerebrovascular stenting. Am J Neuroradiol 2008; 29: 281-285.