Dataset Information

Label free quantitative mass spectrometry identifies platelet degranulation as an early event during platelet storage

ABSTRACT: Shelf life of platelet concentrates (PCs) is limited to up to 7 days due to loss of platelet functionality during storage, commonly referred to as the platelet storage lesion (PSL). To get more insight into the development of the PSL, we used label free quantitative mass spectrometry to identify changes in the platelet proteome during storage. In total 2501 proteins were accurately quantified in 3 biological replicates on at least 1 of the 7 different time-points analyzed. Significant changes in levels of 21 proteins were observed over time. Gene ontology (GO) enrichment analysis of these proteins revealed that the majority of this set was involved in platelet degranulation, secretion and regulated exocytosis. Twelve of these proteins, which included SPARC and von Willebrand factor (VWF), have been shown to reside in α-granules; a significant decline in levels of these proteins was already observed at day 5 of storage. A strongly reduced level of Protein S100A9 (S100A9) which is predominantly localized in the cytoplasm was observed at day 5 of storage. We also monitored changes upon prolonged storage (13-16 days); identifying elevated levels of α-2-macroglobulin, glycogenin and Ig μ chain C region. The ability of platelets to bind and/or endocytose α-2-macroglobulin was confirmed by confocal microscopy. Taken together, our data show that release of α-granules is an early event during platelet storage. In addition, we show that platelets can bind or ingest proteins during storage which may have impact on the hemostatic properties of platelets used for transfusion.

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Platelet, Platelet

SUBMITTER: Maaike Rijkers

LAB HEAD: Jan Voorberg

PROVIDER: PXD005610 | Pride | 2017-09-11

REPOSITORIES: Pride

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Publications

Monitoring storage induced changes in the platelet proteome employing label free quantitative mass spectrometry.

Rijkers Maaike M van den Eshof Bart L BL van der Meer Pieter F PF van Alphen Floris P J FPJ de Korte Dirk D Leebeek Frank W G FWG Meijer Alexander B AB Voorberg Jan J Jansen A J Gerard AJG

Scientific reports 20170908 1

Shelf life of platelet concentrates is limited to 5-7 days due to loss of platelet function during storage, commonly referred to as the platelet storage lesion (PSL). To get more insight into the development of the PSL, we used label free quantitative mass spectrometry to identify changes in the platelet proteome during storage. In total 2501 proteins were accurately quantified in 3 biological replicates on at least 1 of the 7 different time-points analyzed. Significant changes in levels of 21 p ...[more]

PMID: 28887518

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Project description:Platelet concentrates (PCs) represent a blood transfusion product with a major concern for safety as their storage temperature (20-24ºC) allows bacterial growth, and their maximum storage time period (less than a week) precludes complete microbiological testing. Pathogen inactivation technologies (PITs) provide an additional layer of safety to the blood transfusion products from known and unknown pathogens (such as bacteria, viruses and parasites). In this context, PITs (such as Mirasol Pathogen Reduction Technology -PRT-) have been developed and are implemented in many countries. However, several studies have shown in vitro that Mirasol PRT induces a certain level of platelet shape change, hyperactivation, basal degranulation and increased oxidative damage during storage. It has been suggested that Mirasol PRT might accelerate what has been described as the platelet storage lesion (PSL), but supportive molecular signatures have not been obtained. We aimed at dissecting the influence of both variables, i.e. Mirasol PRT and storage time, at the proteome level. We present comprehensive proteomics data analysis of Control PCs and PCs treated with Mirasol PRT at storage days 2, 6 and 8. Our workflow was set to perform proteomics analysis using a gel-free and label-free quantification (LFQ) approach. Semi-quantification was based on LFQ signal intensities of identified proteins using MaxQuant/Perseus software platform. We identified marginal differences between Mirasol PRT and control PCs during storage. However, those significant changes at the proteome level were specifically related to the functional aspects previously described to affect platelets upon Mirasol PRT, and in addition, the effect of Mirasol PRT on the platelet proteome appeared not to be exclusively due to an accelerated or enhanced PSL. In summary, semi-quantitative proteomics allows to discern between proteome changes due to Mirasol PRT or PSL, and proves to be a methodology suitable to phenotype platelets in an unbiased manner, in various physiological contexts.

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