Enzymatically oxygenated phospholipids (eoxPL) formed by lipoxygenases (LOX) and cyclooxygenase (COX) in platelets and leukocytes are pro-coagulant in multiple model systems. However, their generation in arterial thrombotic disease, and how their levels are modulated by common therapies is unknown. Here, eoxPL were first characterized in isolated platelets and leukocytes from an arterial vascular disease cohort, a healthy cohort administered low dose aspirin, and from retrieved human arterial thrombi. In both cohorts, aspirin reduced platelet COX-1-derived eoxPL, while elevating diacyl 12-LOX-derived eoxPL in males, through enhanced Lands cycle esterification. Conversely, P2Y12 inhibition reduced 12-LOX-derived eoxPL in leukocytes. Complex aspirin-dependent gender and seasonal effects on platelet eoxPL were seen in healthy subjects. Limb or coronary (STEMI) thrombi showed a platelet eoxPL signature while carotid thrombi had a white cell profile. Mice genetically lacking leukocyte 12/15-LOX, which are deficient in eoxPL, generated smaller carotid thrombi in vivo. In summary, pro-coagulant eoxPL generation is altered in human arterial vascular disease by commonly used cardiovascular therapies. These changes to the phospholipid composition of blood cells in humans at risk of thrombotic events may be clinically significant where the pro-coagulant membrane plays a central understood role in driving elevated thrombotic risk.

P.C. receives research funding from CSL Behring, Haemonetics Corp, Werfen, and consultancy from CSL Behring.

NCT05604118

This work was supported by the Wellcome Trust (GW4CAT fellowship to M.P 216278/Z/19/Z) and the British Heart Foundation (Programme grant to P.C and V.B.O RG/F/20/110020). Healthy control cohort sample collection was supported by British Heart Foundation (FS/15/45/31603). DB is in receipt of a HCRW NHS Research Time Award. VJT was supported in part by the Welsh Government/EU Ser Cymru Programme. AAH is supported by a grant from Kuwait University.

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Ethical approval, which included informed consent, was from Cardiff University, Schools of Medicine and Dentistry Research Ethics Committee (SMREC16/02). Ethical approval was from Health and Care Research Wales (HCRW, IRAS 243701; REC reference 18/YH/0502). The interventional study is registered on ClinicalTrials.org, NCT05604118.

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