In vivo platelet activation and platelet hyperreactivity in abacavir-treated HIV-infected patients

Summary

Abacavir (ABC) has been associated with ischaemic cardiovascular events in HIV-infected patients, but the pathogenic mechanisms are unknown. Aim of our study was to assess whether ABC induces in vivo platelet activation and ex vivo platelet hyper-reactivity. In a retrospective, case-control study, in vivo platelet activation markers were measured in 69 HIV-infected patients, before starting therapy and after 6–12 months of either ABC (n=35) or tenofovir (TDF) (n=34), and compared with those from 20 untreated HIV-infected patients. A subgroup of patients was restudied after 28–34 months for ex vivo platelet reactivity. In vivo platelet activation markers were assessed by ELISA or flow cytometry, ex vivo platelet reactivity by light transmission aggregometry (LTA) and PFA-100®. The in vitro effects of the ABC metabolite, carbovir triphosphate, on aggregation and intra-platelet cGMP were also studied. sPLA2, sPsel and sGPV increased significantly 6–12 months after the beginning of ABC, but not of TDF or of no treatment. Ex vivo platelet function studies showed enhanced LTA, shorter PFA-100® C/ADP closure time and enhanced platelet expression of P-sel and CD40L in the ABC group. The intake of ABC blunted the increase of intraplatelet cGMP induced by nitric oxide (NO) and acutely enhanced collagen-induced aggregation. Preincubation of control platelets with carbovir triphosphate in vitro enhanced platelet aggregation and blunted NO-induced cGMP elevation. In conclusion, treatment with ABC enhances in vivo platelet activation and induces platelet hyperreactivity by blunting the inhibitory effects of NO on platelets. These effects may lead to an increase of ischaemic cardiovascular events.

Keywords

Abacavir - cardiovascular risk - guanylyl cyclase - nitric oxide - platelet aggregation

^* These authors contributed equally to this work.

• References

• 1 Porter K, Babiker A, Bhaskaran K. et al. Determinants of survival following HIV-1 seroconversion after the introduction of HAART. Lancet 2003; 362: 1267-1274.
• 2 Baker JV, Lundgren JD. Cardiovascular implications from untreated human immunodeficiency virus infection. Eur Heart J 2011; 32: 945-951.
• 3 Gresele P, Falcinelli E, Sebastiano M. et al. Endothelial and platelet function alterations in HIV-infected patients. Thromb Res 2012; 129: 301-8.
• 4 Choi AI, Vittinghoff E, Deeks SG. et al. Cardiovascular risks associated with abacavir and tenofovir exposure in HIV-infected persons. AIDS 2011; 25: 1289-1298.
• 5 D: A: D Study Group. Sabin CA, Worm SW. et al. Use of nucleoside reverse transcriptase inhibitors and risk of myocardial infarction in HIV-infected patients enrolled in the D: A: D study: a multicohort collaboration. Lancet 2008; 371: 1417-1426.
• 6 The SMART/INSIGHT and the D: A: D study groups. Use of nucleoside reverse transcriptase inhibitors and risk of myocardial infarction in HIV infected patients. AIDS 2008; 22: F17-24.
• 7 Ribaudo HJ, Benson CA, Zheng Y. et al. No risk of myocardial infarction associated with initial antiretroviral treatment containing abacavir: short and longterm results from ACTG A5001/ALLRT. Clin Infect Dis 2011; 52: 929-940.
• 8 Saumoy M, Ordoñez-Llanos J, Martínez E. et al. Low-density lipoprotein size and lipoprotein-associated phospholipase A2 in HIV-infected patients switching to abacavir or tenofovir. Antivir Ther 2011; 16: 459-468.
• 9 Packard RR, Libby P. Inflammation in atherosclerosis: from vascular biology to biomarker discovery and risk prediction. Clin Chem 2008; 54: 24-38.
• 10 Reriani MK, Lerman LO, Lerman A. Endothelial function as a functional expression of cardiovascular risk factors. Biomark Med 2010; 04: 351-360.
• 11 Gresele P, Momi S, Migliacci R. Endothelium, venous thromboembolism and ischaemic cardiovascular events. Thromb Haemost 2010; 103: 56-61.
• 12 Hsue PY, Hunt PW, Wu Y. et al. Association of abacavir and impaired endothelial function in treated and suppressed HIV-infected patients. AIDS 2009; 23: 2021-2027.
• 13 Behrens GM, Reiss P. Abacavir and cardiovascular risk. Curr Opin Infect Dis 2010; 23: 9-14.
• 14 Davì G, Patrono C. Platelet activation and atherothrombosis. N Engl J Med 2007; 357: 2482-2494.
• 15 Harrison P, Keeling D. Platelet hyperactivity and risk of recurrent thrombosis. J Thromb Haemost 2006; 04: 2544-2546.
• 16 Frossard M, Fuchs I, Leitner JM. et al. Platelet function predicts myocardial damage in patients with acute myocardial infarction. Circulation 2004; 110: 1392-1397.
• 17 Holme PA, Muller F, Solum NO. et al. Enhanced activation of platelets with abnormal release of RANTES in human immunodeficiency virus type 1 infection. FASEB J 1998; 12: 79-89.
• 18 Corrales-Medina VF, Simkins J, Chirinos JA. et al. Increased levels of platelet microparticles in HIV-infected patients with good response to highly active antiretroviral therapy. J Acquir Immune Defic Syndr 2010; 54: 217-218.
• 19 Landrø L, Ueland T, Otterdal K. et al. Persistently raised plasma levels of platelet-derived inflammatory mediators in HIV-infected patients during highly active anti-retroviral therapy. J Thromb Haemost 2011; 09: 1075-1077.
• 20 Satchell CS, Cotter AG, O’Connor EF. et al. Platelet function and HIV: a casecontrol study. AIDS 2010; 24: 649-657.
• 21 Mallal S, Phillips E, Carosi G. et al. HLA-B*5701 screening for hypersensitivity to abacavir. N Engl J Med 2008; 358: 568-579.
• 22 Marroni M, Gresele P, Landonio G. et al. Interferon-alpha is effective in the treatment of HIV-1-related, severe, zidovudine-resistant thrombocytopenia. A prospective, placebo-controlled, double-blind trial. Ann Intern Med 1994; 121: 423-429.
• 23 Francisci D, Giannini S, Baldelli F. et al. HIV type 1 infection, and not shortterm HAART, induces endothelial dysfunction. AIDS 2009; 23: 589-596.
• 24 Giannini S, Falcinelli E, Bury L. et al. Interaction with damaged vessel wall in vivo in humans induces platelets to express CD40L resulting in endothelial activation. No effect of aspirin intake. Am J Physiol Heart Circ Physiol 2011; 300: H2072-2079.
• 25 Gresele P, Falcinelli E, Loffredo F. et al. Platelets release matrix metalloproteinase-2 in the coronary circulation of patients with acute coronary syndromes: possible role in sustained platelet activation. Eur Heart J 2011; 32: 316-325.
• 26 Falcinelli E, Guglielmini G, Torti M. et al. Intraplatelet signaling mechanisms of the priming effect of matrix metalloproteinase-2 on platelet aggregation. J Thromb Haemost 2005; 03: 2526-2535.
• 27 Giannini S, Mezzasoma AM, Leone M. et al. Laboratory diagnosis and monitoring of desmopressin treatment of von Willebrand’s disease by flow cytometry. Haematologica 2007; 92: 1647-1654.
• 28 Ciferri S, Emiliani C, Guglielmini G. et al. Platelets release their lysosomal content in vivo in humans upon activation. Thromb Haemost 2000; 83: 157-164.
• 29 Knight CJ, Panesar M, Wright C. et al. Altered platelet function detected by flow cytometry. Effects of coronary artery disease and age. Arterioscler Thromb Vasc Biol 1997; 17: 2044-2053.
• 30 Rossiello MR, Momi S, Caracchini R. et al. A novel nitric oxide-releasing statin derivative exerts an antiplatelet/antithrombotic activity and inhibits tissue factor expression. J Thromb Haemost 2005; 03: 2554-2562.
• 31 D’Avolio A, Sciandra M, Siccardi M. et al. A new assay based on solid-phase extraction procedure with LC-MS to measure plasmatic concentrations of tenofovir and emtricitabine in HIV infected patients. J Chromatogr Sci 2008; 46: 524-528.
• 32 Satchell CS, O’Halloran JA, Cotter AG. et al. Increased platelet reactivity in HIV-1-Infected patients receiving abacavir-containing antiretroviral therapy. J Infect Dis 2011; 204: 1202-1210.
• 33 Blann AD, Lip GYH, Beevers DG. et al. Soluble P-selectin in atherosclerosis: a comparison with endothelial cell and platelet markers. Thromb Haemost 1997; 77: 1077-1080.
• 34 Fateh-Moghadam S, Li Z, Ersel S. et al. Platelet degranulation is associated with progression of intima-media thickness of the common carotid artery in patients with diabetes mellitus type 2. Arterioscler Thromb Vasc Biol 2005; 25: 1299-1303.
• 35 André P, Nannizzi-Alaimo L, Prasad SK. Platelet-derived CD40L: the switchhitting player of cardiovascular disease. Circulation 2002; 106: 896-899.
• 36 Heeschen C, Dimmeler S, Hamm CW. et al. Soluble CD40 ligand in acute coronary syndromes. N Engl J Med 2003; 348: 1104-11.
• 37 Atalar E, Haznedaroglu IC, Kilic H. et al. Increased soluble glycoprotein V concentration during the acute onset of unstable angina pectoris in association with chronic cigarette smoking. Platelets 2005; 16: 329-333.
• 38 Ruggeri ZM. Mechanisms initiating platelet thrombus formation. Thromb Haemost 1997; 78: 611-616.
• 39 Gresele P, Guglielmini G, De Angelis M. et al. Acute, short-term hyperglycemia enhances shear stress-induced platelet activation in patients with type II diabetes mellitus. J Am Coll Cardiol 2003; 41: 1013-1020.
• 40 Baum PD, Sullam PM, Stoddart CA. et al. Abacavir increases platelet reactivity via competitive inhibition of soluble guanylyl cyclase. AIDS 2011; 25: 2243-2248.
• 41 Gresele P, Marzotti S, Guglielmini G. et al. Hyperglycemia-induced platelet activation in type 2 diabetes is resistant to aspirin but not to a nitric oxide-donating agent. Diabetes Care 2010; 33: 1262-1268.
• 42 Moyle G, Boffito M, Fletcher C. et al. Steady-state pharmacokinetics of abacavir in plasma and intracellular carbovir triphosphate following administration of abacavir at 600 milligrams once daily and 300 milligrams twice daily in human immunodeficiency virus-infected subjects. Antimicrob Agents Chemother 2009; 53: 1532-1538.
• 43 Rajendran S, Chirkov YY. Platelet hyperaggregability: impaired responsiveness to nitric oxide (“platelet NO resistance”) as a therapeutic target. Cardiovasc Drugs Ther 2008; 22: 193-203.
• 44 Willoughby SR, Stewart S, Holmes AS. et al. Platelet nitric oxide responsiveness: a novel prognostic marker in acute coronary syndromes. Arterioscler Thromb Vasc Biol 2005; 25: 2661-2666.
• 45 Lev EI, Alviar CL, Arikan ME. et al. Platelet reactivity in patients with subacute stent thrombosis compared with non-stent-related acute myocardial infarction. Am Heart J 2007; 153: 41 e1–6.
• 46 D’Ascenzo F, Cerrato E, Biondi-Zoccai G. et al. Acute coronary syndromes in human immunodeficiency virus patients: a meta-analysis investigating adverse event rates and the role of antiretroviral therapy. Eur Heart J 2011; 33: 875-880.
• 47 Francisci D, Falcinelli E, Belfiori B. et al. HIV infection-induced endothelial dysfunction is partially corrected by tenofovir but less by abacavir. AIDS Patient Care and STDs 2011; 25: 567-569.
• 48 Schäfer A, Alp NJ, Cai S. et al. Reduced vascular NO bioavailability in diabetes increases platelet activation in vivo. Arterioscler Thromb Vasc Biol 2004; 24: 1720-1726.