Thijs C van Holten1, Onno B Bleijerveld2, Patrick Wijten3, Philip G de Groot1, Albert J R Heck3, Arjan D Barendrecht1, Tesy H Merkx1, Arjen Scholten4, Mark Roest5. 1. Laboratory for Clinical Chemistry and Haematology, UMC Utrecht, Utrecht, The Netherlands. 2. Laboratory of Experimental Cardiology, UMC Utrecht, Utrecht, The Netherlands Biomolecular Mass Spectrometry and Proteomics, Utrecht University, Utrecht, The Netherlands Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands Netherlands Proteomics Center, Utrecht, The Netherlands. 3. Biomolecular Mass Spectrometry and Proteomics, Utrecht University, Utrecht, The Netherlands Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands Netherlands Proteomics Center, Utrecht, The Netherlands. 4. Biomolecular Mass Spectrometry and Proteomics, Utrecht University, Utrecht, The Netherlands Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands Netherlands Proteomics Center, Utrecht, The Netherlands a.scholten@uu.nl mroest@umcutrecht.nl. 5. Laboratory for Clinical Chemistry and Haematology, UMC Utrecht, Utrecht, The Netherlands a.scholten@uu.nl mroest@umcutrecht.nl.
Abstract
AIMS: Platelets are a natural source of growth factors, cytokines and chemokines, that regulate angiogenesis and inflammation. It has been suggested that differential release of pro- and anti-angiogenic growth factors from platelet α-granules by protease-activated receptors (PAR) 1 and 4 may be important for the regulation of angiogenesis. We aimed to compare the releasates of unstimulated platelets with PAR-1- and PAR-4-stimulated platelets. METHODS AND RESULTS: The release of β-thromboglobulin, platelet factor (PF)-4, thrombospondin, platelet-derived growth factor (PDGF)-A/B, regulated and normal T-cell expressed and secreted (RANTES/CCL5), endostatin, CXCL12, and vascular endothelial growth factor (VEGF) was measured with enzyme-linked immunosorbent assay (ELISA). Mass spectrometry (MS)-based quantitative proteomics identified 93 proteins from platelets stimulated with PAR-1 and PAR-4. A strong correlation between the factors released after either stimulus was observed (Spearman's r 0.94, P < 0.001). Analysis with ELISA showed that stimulation with PAR-1 or PAR-4 lead to non-differential release of β-thromboglobulin, PF-4, thrombospondin, PDGF-A/B, RANTES/CCL5, endostatin, CXCL12, and VEGF. Release of thrombospondin was slightly lower after PAR-1 stimulation (7.2 μg/mL), compared with PAR-4 induced release (9.8 μg/mL; P < 0.05). CONCLUSIONS: Both ELISA on established α-granule proteins and MS-based quantitative proteomics showed that the most abundant α-granule proteins are released in similar quantities from platelets after stimulation with either PAR-1 or PAR-4. Our findings provide evidence against the hypothesis that PAR-1 and PAR-4 stimulation of platelets trigger differential release of alpha-granule, but further studies are needed to draw conclusions for physiological conditions. Published on behalf of the European Society of Cardiology. All rights reserved.
AIMS: Platelets are a natural source of growth factors, cytokines and chemokines, that regulate angiogenesis and inflammation. It has been suggested that differential release of pro- and anti-angiogenic growth factors from platelet α-granules by protease-activated receptors (PAR) 1 and 4 may be important for the regulation of angiogenesis. We aimed to compare the releasates of unstimulated platelets with PAR-1- and PAR-4-stimulated platelets. METHODS AND RESULTS: The release of β-thromboglobulin, platelet factor (PF)-4, thrombospondin, platelet-derived growth factor (PDGF)-A/B, regulated and normal T-cell expressed and secreted (RANTES/CCL5), endostatin, CXCL12, and vascular endothelial growth factor (VEGF) was measured with enzyme-linked immunosorbent assay (ELISA). Mass spectrometry (MS)-based quantitative proteomics identified 93 proteins from platelets stimulated with PAR-1 and PAR-4. A strong correlation between the factors released after either stimulus was observed (Spearman's r 0.94, P < 0.001). Analysis with ELISA showed that stimulation with PAR-1 or PAR-4 lead to non-differential release of β-thromboglobulin, PF-4, thrombospondin, PDGF-A/B, RANTES/CCL5, endostatin, CXCL12, and VEGF. Release of thrombospondin was slightly lower after PAR-1 stimulation (7.2 μg/mL), compared with PAR-4 induced release (9.8 μg/mL; P < 0.05). CONCLUSIONS: Both ELISA on established α-granule proteins and MS-based quantitative proteomics showed that the most abundant α-granule proteins are released in similar quantities from platelets after stimulation with either PAR-1 or PAR-4. Our findings provide evidence against the hypothesis that PAR-1 and PAR-4 stimulation of platelets trigger differential release of alpha-granule, but further studies are needed to draw conclusions for physiological conditions. Published on behalf of the European Society of Cardiology. All rights reserved.
Authors: Monika Haemmerle; Rebecca L Stone; David G Menter; Vahid Afshar-Kharghan; Anil K Sood Journal: Cancer Cell Date: 2018-04-12 Impact factor: 31.743
Authors: Lauren G Poole; Veronica L Massey; Deanna L Siow; Edilson Torres-Gonzáles; Nikole L Warner; James P Luyendyk; Jeffrey D Ritzenthaler; Jesse Roman; Gavin E Arteel Journal: Am J Respir Cell Mol Biol Date: 2017-09 Impact factor: 6.914
Authors: Kellie R Machlus; Kelly E Johnson; Rajesh Kulenthirarajan; Jodi A Forward; Mason D Tippy; Thomas S Soussou; Saleh H El-Husayni; Stephen K Wu; Suming Wang; Randolph S Watnick; Joseph E Italiano; Elisabeth M Battinelli Journal: Blood Date: 2015-12-08 Impact factor: 22.113
Authors: Frauke Swieringa; Fiorella A Solari; Albert Sickmann; Mattia Frontini; Johan W M Heemskerk; Jingnan Huang; Isabella Provenzale; Luigi Grassi; Ilaria De Simone; Constance C F M J Baaten; Rachel Cavill Journal: Sci Rep Date: 2021-06-11 Impact factor: 4.379