title: Necrotic platelets provide a procoagulant surface during thrombosis.
abstract: A subpopulation of platelets fulfills a procoagulant role in hemostasis and thrombosis by enabling the thrombin burst required for fibrin formation and clot stability at the site of vascular injury. Excess procoagulant activity is linked with pathological thrombosis. The identity of the procoagulant platelet has been elusive. The cell death marker 4-[N-(S-glutathionylacetyl)amino]phenylarsonous acid (GSAO) rapidly enters a subpopulation of agonist-stimulated platelets via an organic anion-transporting polypeptide and is retained in the cytosol through covalent reaction with protein dithiols. Labeling with GSAO, together with exposure of P-selectin, distinguishes necrotic from apoptotic platelets and correlates with procoagulant potential. GSAO(+) platelets form in occluding murine thrombi after ferric chloride injury and are attenuated with megakaryocyte-directed deletion of the cyclophilin D gene. These platelets form a procoagulant surface, supporting fibrin formation, and reduction in GSAO(+) platelets is associated with reduction in platelet thrombus size and fibrin formation. Analysis of platelets from human subjects receiving aspirin therapy indicates that these procoagulant platelets form despite aspirin therapy, but are attenuated by inhibition of the necrosis pathway. These findings indicate that the major subpopulation of platelets involved in fibrin formation are formed via regulated necrosis involving cyclophilin D, and that they may be targeted independent of platelet activation.
authors.affiliation: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales, Australia;
authors.forename: Vu Minh
authors.initials: VM
authors.lastname: Hua
authors.affiliation: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales, Australia;
authors.forename: Latasha
authors.initials: L
authors.lastname: Abeynaike
authors.affiliation: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales, Australia;
authors.forename: Elias
authors.initials: E
authors.lastname: Glaros
authors.affiliation: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales, Australia;
authors.forename: Heather
authors.initials: H
authors.lastname: Campbell
authors.affiliation: Haematology, Institute of Clinical Pathology and Medical Research, Westmead, New South Wales, Australia;
authors.forename: Leonardo
authors.initials: L
authors.lastname: Pasalic
authors.affiliation: The Centenary Institute, Newtown, Sydney, New South Wales, Australia; National Health and Medical Research Council Clinical Trials Centre, University of Sydney, Sydney, New South Wales, Australia;
authors.forename: Philip J
authors.identifier: http://orcid.org/0000-0001-6486-2863
authors.initials: PJ
authors.lastname: Hogg
authors.affiliation: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales, Australia; Haematology, Concord Repatriation and General Hospital, Sydney, New South Wales, Australia; and ANZAC Research Institute, Concord, New South Wales, Australia.
authors.forename: Vivien M Y
authors.initials: VM
authors.lastname: Chen
mesh_terms: D000818:Animals; D001152:Arsenicals; D015415:Biomarkers / Q000032:analysis; D001792:Blood Platelets / Q000378:metabolism*; D002478:Cells, Cultured; D021983:Cyclophilins / Q000378:metabolism; D005434:Flow Cytometry; D005978:Glutathione / Q000031:analogs & derivatives; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D009336:Necrosis / Q000378:metabolism; D015539:Platelet Activation / Q000502:physiology*; D013927:Thrombosis / Q000378:metabolism*; D000097322:Peptidyl-Prolyl Isomerase D
chemical_list: D001152:Arsenicals; D015415:Biomarkers; D021983:Cyclophilins; D005978:Glutathione; C426645:4-(N-(S-glutathionylacetyl)amino)phenylarsenoxide; C114456:PPID protein, human; D000097322:Peptidyl-Prolyl Isomerase D