Succinate reverses in-vitro platelet inhibition by acetylsalicylic acid and P2Y receptor antagonists.

TY - JOUR

T1 - Succinate reverses in-vitro platelet inhibition by acetylsalicylic acid and P2Y receptor antagonists.

AU - Spath, Brigitte

AU - Hansen, Arne

AU - Bokemeyer, Carsten

AU - Langer, Florian

PY - 2012

Y1 - 2012

N2 - High on-treatment platelet reactivity has been associated with adverse cardiovascular events in patients receiving anti-platelet agents, but the molecular mechanisms underlying this phenomenon remain incompletely understood. Succinate, a citric acid cycle intermediate, is released into the circulation under conditions of mitochondrial dysfunction due to hypoxic organ damage, including sepsis, stroke, and myocardial infarction. Because the G protein-coupled receptor (GPCR) for succinate, SUCNR1 (GPR91), is present on human platelets, we hypothesized that succinate-mediated platelet stimulation may counteract the pharmacological effects of cyclooxygenase-1 and ADP receptor antagonists. To test this hypothesis in a controlled in-vitro study, washed platelets from healthy donors were treated with acetylsalicylic acid (ASA) or small-molecule P2Y(1) or P2Y(12) inhibitors and subsequently analyzed by light transmittance aggregometry using arachidonic acid (AA), ADP and succinate as platelet agonists. Aggregation in response to succinate alone was highly variable with only 29% of donors showing a (mostly delayed) platelet response. In contrast, succinate reproducibly and concentration-dependently (10-1000?µM) enhanced platelet aggregation in response to low concentrations of exogenous ADP. Furthermore, while succinate alone had no effect in the presence of platelet inhibitors, responsiveness of platelets to ADP after pretreatment with P2Y(1) or P2Y(12) antagonists was fully restored, when platelets were co-stimulated with 100?µM succinate. Similarly, succinate completely (at 1000?µM) or partially (at 100?µM) reversed the inhibitory effect of ASA on AA-induced platelet aggregation. In contrast, succinate failed to restore platelet responsiveness in the presence of both ASA and the P2Y(12) antagonist, suggesting that concomitant signaling via different GPCRs was required. Essentially identical results were obtained, when flow cytometric analysis of surface CD62P expression was used as a different readout for platelet activation. In summary, extracellular succinate may have a co-stimulatory role in platelet aggregation and, by (partially) antagonizing the effects of platelet inhibitors, may contribute to the inter-individual variability frequently observed in platelet function testing.

AB - High on-treatment platelet reactivity has been associated with adverse cardiovascular events in patients receiving anti-platelet agents, but the molecular mechanisms underlying this phenomenon remain incompletely understood. Succinate, a citric acid cycle intermediate, is released into the circulation under conditions of mitochondrial dysfunction due to hypoxic organ damage, including sepsis, stroke, and myocardial infarction. Because the G protein-coupled receptor (GPCR) for succinate, SUCNR1 (GPR91), is present on human platelets, we hypothesized that succinate-mediated platelet stimulation may counteract the pharmacological effects of cyclooxygenase-1 and ADP receptor antagonists. To test this hypothesis in a controlled in-vitro study, washed platelets from healthy donors were treated with acetylsalicylic acid (ASA) or small-molecule P2Y(1) or P2Y(12) inhibitors and subsequently analyzed by light transmittance aggregometry using arachidonic acid (AA), ADP and succinate as platelet agonists. Aggregation in response to succinate alone was highly variable with only 29% of donors showing a (mostly delayed) platelet response. In contrast, succinate reproducibly and concentration-dependently (10-1000?µM) enhanced platelet aggregation in response to low concentrations of exogenous ADP. Furthermore, while succinate alone had no effect in the presence of platelet inhibitors, responsiveness of platelets to ADP after pretreatment with P2Y(1) or P2Y(12) antagonists was fully restored, when platelets were co-stimulated with 100?µM succinate. Similarly, succinate completely (at 1000?µM) or partially (at 100?µM) reversed the inhibitory effect of ASA on AA-induced platelet aggregation. In contrast, succinate failed to restore platelet responsiveness in the presence of both ASA and the P2Y(12) antagonist, suggesting that concomitant signaling via different GPCRs was required. Essentially identical results were obtained, when flow cytometric analysis of surface CD62P expression was used as a different readout for platelet activation. In summary, extracellular succinate may have a co-stimulatory role in platelet aggregation and, by (partially) antagonizing the effects of platelet inhibitors, may contribute to the inter-individual variability frequently observed in platelet function testing.

KW - Humans

KW - Male

KW - Female

KW - Aspirin/pharmacology

KW - Blood Platelets/cytology/metabolism

KW - Platelet Aggregation/drug effects

KW - Platelet Aggregation Inhibitors/pharmacology

KW - Purinergic P2Y Receptor Agonists/pharmacology

KW - Receptors, G-Protein-Coupled/metabolism

KW - Receptors, Purinergic P2Y12/metabolism

KW - Succinic Acid/pharmacology

KW - Humans

KW - Male

KW - Female

KW - Aspirin/pharmacology

KW - Blood Platelets/cytology/metabolism

KW - Platelet Aggregation/drug effects

KW - Platelet Aggregation Inhibitors/pharmacology

KW - Purinergic P2Y Receptor Agonists/pharmacology

KW - Receptors, G-Protein-Coupled/metabolism

KW - Receptors, Purinergic P2Y12/metabolism

KW - Succinic Acid/pharmacology

M3 - SCORING: Journal article

VL - 23

SP - 60

EP - 68

JO - PLATELETS

JF - PLATELETS

SN - 0953-7104

IS - 1

M1 - 1

ER -