NOD2-mediated P2Y12 upregulation increases platelet activation and thrombosis in sepsis

Abstract

Background: Platelets from septic patients exhibit increased reactivity. However, the underlying mechanism of sepsis-induced platelet hyperactivity is still not completely understood.

Objective: P2Y₁₂ is a central receptor for platelet activation. In this study, we investigated the role of platelet P2Y₁₂ in platelet hyperactivity during sepsis.

Methods: We measured platelet P2Y₁₂ expression and aggregation in response to ADP in septic patients and cecal ligation and puncture (CLP)-treated mice. We also detected the downstream signaling of P2Y₁₂ in resting platelets from patients and mice with sepsis. The role of nucleotide-binding oligomerization domain 2 (NOD2)/RIP2/NF-κB/P65 pathway in sepsis-induced platelet P2Y₁₂ high expression was also investigated. Finally, we compared the antiplatelet and antithrombotic effects of clopidogrel, prasugrel, and ticagrelor in experimental sepsis in mice and rats.

Results: Compared to healthy subjects, platelets from septic patients exhibit P2Y₁₂ hyperactivity and higher P2Y₁₂ expression. pAkt is enhanced and pVASP is impaired in resting platelets from the patients, indicating the constitutive activation of platelet P2Y₁₂ receptor. Mouse sepsis model recapitulates the findings in septic patients. NOD2 deficiency attenuates sepsis-induced platelet P2Y₁₂ high expression, hyperactivity, and thrombosis. Prasugrel and ticagrelor are potent P2Y₁₂ inverse agonists, and exhibit superior antiplatelet and antithrombotic efficacy over clopidogrel in mice and rats with sepsis.

Conclusions: NOD2 activation upregulates platelet P2Y₁₂ expression, which is constitutively activated and contributes to platelet hyperactivity in septic status. Compared to clopidogrel, prasugrel and ticagrelor are potent P2Y₁₂ inverse agonists with superior antiplatelet and antithrombotic efficacy in experimental sepsis.

Keywords: Inverse agonists; NOD2; P2Y(12); Platelet activation; Sepsis.