The canine activated platelet secretome (CAPS): A translational model of thrombin‐evoked platelet activation response

• Main Authors: Signe E. Cremer, James L. Catalfamo, Robert Goggs, Stefan E. Seemann, Annemarie T. Kristensen, Paulina B. Szklanna, Patricia B. Maguire, Marjory B. Brooks
• Format: Article
• Language: English
• Published: Wiley 2021-01-01
• Series: Research and Practice in Thrombosis and Haemostasis
• Subjects: dog; platelet; proteomics; releasate; secretion
• Online Access: https://doi.org/10.1002/rth2.12450

Abstract Background Domestic dogs represent a translational animal model to study naturally occurring human disease. Proteomics has emerged as a promising tool for characterizing human platelet pathophysiology; thus a detailed characterization of the core canine activated platelet secretome (CAPS) will enhance utilization of the canine model. The objectives of this study were development of a robust, high throughput, label‐free approach for proteomic identification and quantification of the canine platelet (i) thrombin releasate proteins, and (ii) the protein subgroup that constitutes CAPS. Methods Platelets were isolated from 10 healthy dogs and stimulated with 50 nmol/L of γ‐thrombin or saline. Proteins were in‐solution trypsin‐digested and analyzed by nano–liquid chromatography–tandem spectrometry. Core releasate proteins were defined as those present in 10 of 10 dogs, and CAPS defined as releasate proteins with a significantly higher abundance in stimulated versus saline controls (corrected P < .05). Results A total of 2865 proteins were identified; 1126 releasate proteins were present in all dogs, 650 were defined as CAPS. Among the differences from human platelets were a canine lack of platelet factor 4 and vascular endothelial growth factor C, and a 10‐ to 20‐fold lower concentration of proteins such as haptoglobin, alpha‐2 macroglobulin, von Willebrand factor, and amyloid‐beta A4. Twenty‐eight CAPS proteins, including cytokines, adhesion molecules, granule proteins, and calcium regulatory proteins have not previously been attributed to human platelets. Conclusions CAPS proteins represent a robust characterization of a large animal platelet secretome and a novel tool to model platelet physiology, pathophysiology, and to identify translational biomarkers of platelet‐mediated disease.