Abstract Number: LPB0083
Meeting: ISTH 2021 Congress
Background: Despite the importance of ex vivo studies of platelets in health and disease, the progressive impact of pre-analytical blood processing conditions on platelet function and composition remains minimally characterized.
Aims: To determine the effects of commonly utilized blood draw anticoagulants, as well as processing time, on ex vivo platelet proteome and function.
Methods: Whole blood was collected from four healthy human donors into four different anticoagulants, including sodium citrate, acid citrate dextrose (ACD), EDTA, and heparin. Platelet function was tracked in anticoagulated blood over 120 hours with biochemical and cellular assays. Whole proteome profiles of platelets purified at 1 hour and 24 hours after blood draw, were determined through a peptide tandem mass tag (TMT) labeling and multiplex mass spectrometry approach.
Results: Collection of blood into heparin, and to a lesser extent sodium citrate, significantly increased platelet-platelet and platelet-monocyte aggregate formation within 1 hour of blood draw. A rapid release of platelet-derived extracellular vesicles was also observed for heparinized blood, whereas a distinct increase in platelet surface P-selectin exposure was noted for platelets in EDTA blood 5 hours after collection. Multiplex TMT proteomics identified 3,357 proteins spanning a dynamic range of 5 orders of magnitude in all platelet samples, where, the duration of blood storage before platelet purification was a strong driver of whole platelet proteome changes associated with metabolism and exocytosis. Compared to platelets from ACD-anticoagulated blood, EDTA platelets showed increased levels of complement C1r and ficolin 3 proteins. Platelets from heparinized blood contained high levels of histone proteins and neutrophil-related enzymes. The Association of NET products and platelets was confirmed by flow cytometry and immunofluorescence staining.
Conclusions: This study establishes time-dependent and anticoagulant-associated ex vivo effects on the platelet proteo-sequestrome that may confound characterizations of platelet function in health and disease.
To cite this abstract in AMA style:Tassi Yunga S, Gowers A, Melrose A, Fitzgerald M, Rajendran A, Reddy A, Wilmarth P, David L, McCarty O, Aslan J. Temporal Assessment of Platelet Proteome and fFnction during ex vivo Anticoagulation [abstract]. Res Pract Thromb Haemost. 2021; 5 (Suppl 2). https://abstracts.isth.org/abstract/temporal-assessment-of-platelet-proteome-and-ffnction-during-ex-vivo-anticoagulation/. Accessed December 11, 2023.
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ISTH Congress Abstracts - https://abstracts.isth.org/abstract/temporal-assessment-of-platelet-proteome-and-ffnction-during-ex-vivo-anticoagulation/