title: Secretome profiling of apheresis platelet supernatants during routine storage via antibody-based microarray.
abstract: Platelet storage lesions (PSLs) occur during platelet concentrate (PC) storage. Adverse transfusion reactions (ATRs) have been demonstrated to be more frequent in older PCs and removal of the supernatant prior to transfusion reduces their occurrence. Proteomic profiling of PC supernatants was thus performed to identify proteins associated with PSLs and ATRs. Twenty-four PCs were investigated daily from day 0 to day 9 for platelet pre-activation (PPA), platelet-derived extracellular vesicles (PEVs), and platelet function. Using antibody microarrays, 673 extracellular proteins were analysed in PC supernatants on days 0, 3, 5, 7, and 9. During 5days of storage, PPA and PEVs continuously increased (P<0.0001). Platelet function was observed to remain stable within the first 5days (P=0.1751) and decreased thereafter. Comparison of all time points to day 0 revealed the identification of 136 proteins that were significantly changed in abundance during storage, of which 72 were expressed by platelets. Network analysis identified these proteins to be predominantly associated with exosomes (P=4.61×10-8, n=45 genes) and two clusters with distinct functions were found with one being associated with haemostasis and the other with RNA binding. These findings may provide an explanation for ATRs.
  
  
  
  Changes in platelet concentrate (PC) supernatants during storage have been so far only poorly addressed and high abundant proteins burden the identification of quantitative changes in the secretome. We applied a high-throughput antibody microarray allowing for the sensitive quantification of 673 extracellular factors. PCs account for the highest number of adverse transfusion reactions (ATRs). ATRs have been demonstrated to be more frequent in older PCs and removal of the supernatant prior to transfusion reduces their occurrence. Comprehensive interpretation of the changing proteins in the secretome during platelet storage under blood banking conditions may help to identify mechanisms leading to the occurrence of adverse transfusion reactions.
authors.affiliation: Institute of Transfusion Medicine, Charité University Medicine Berlin, Augustenburger Platz 1, 13349 Berlin, Germany. Electronic address: julian.milz@charite.de.
authors.forename: Julian
authors.initials: J
authors.lastname: Kamhieh-Milz
authors.affiliation: Division of Functional Genome Analysis, Deutsches Krebsforschungszentrum (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany; Kurdistan Institution for Strategic Studies and Scientific Research, Gullabax 335, Shorsh St., Sulaimani, Kurdistan Region, Iraq.
authors.forename: Shakhawan A
authors.initials: SA
authors.lastname: Mustafa
authors.affiliation: Institute of Transfusion Medicine, Charité University Medicine Berlin, Augustenburger Platz 1, 13349 Berlin, Germany.
authors.forename: Viktor
authors.initials: V
authors.lastname: Sterzer
authors.affiliation: Institute of Transfusion Medicine, Charité University Medicine Berlin, Augustenburger Platz 1, 13349 Berlin, Germany.
authors.forename: Hatice
authors.initials: H
authors.lastname: Celik
authors.affiliation: Institute of Transfusion Medicine, Charité University Medicine Berlin, Augustenburger Platz 1, 13349 Berlin, Germany.
authors.forename: Sahime
authors.initials: S
authors.lastname: Keski
authors.affiliation: Institute of Transfusion Medicine, Charité University Medicine Berlin, Augustenburger Platz 1, 13349 Berlin, Germany.
authors.forename: Omid
authors.initials: O
authors.lastname: Khorramshahi
authors.affiliation: Institute of Transfusion Medicine, Charité University Medicine Berlin, Augustenburger Platz 1, 13349 Berlin, Germany.
authors.forename: Kamran
authors.initials: K
authors.lastname: Movassaghi
authors.affiliation: Division of Functional Genome Analysis, Deutsches Krebsforschungszentrum (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
authors.forename: Jörg D
authors.initials: JD
authors.lastname: Hoheisel
authors.affiliation: Division of Functional Genome Analysis, Deutsches Krebsforschungszentrum (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
authors.forename: Mohamed S S
authors.initials: MS
authors.lastname: Alhamdani
authors.affiliation: Institute of Transfusion Medicine, Charité University Medicine Berlin, Augustenburger Platz 1, 13349 Berlin, Germany.
authors.forename: Abdulgabar
authors.initials: A
authors.lastname: Salama
mesh_terms: D000906:Antibodies / Q000378:metabolism*; D001792:Blood Platelets / Q000378:metabolism*; D001793:Blood Preservation / Q000379:methods; D064368:Healthy Volunteers; D006801:Humans; D010983:Plateletpheresis; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D013997:Time Factors; D046888:Tissue Array Analysis / Q000379:methods*
chemical_list: D000906:Antibodies; D020543:Proteome
keywords: Antibody microarray; Exosomes; Intracellular calcium; Platelet storage lesions; Platelet-derived extracellular vesicles