Proteome and functional decline as platelets age in the circulation

Anucleate platelets circulate in the blood of healthy individuals for approximately 7-10 days during which time their protein composition may change. We hypothesized such changes would be linked to altered structure and function. Here, we separated platelets of different ages based on mRNA content and characterised them using proteomics, immunofluorescence and functional assays. Total protein content was 45±5% (n=4) lower in old platelets compared to young platelets. Predictive proteomic pathway analysis identified associations with 28 biological processes, notably increased haemostasis in young platelets and apoptosis in old platelets. Further studies confirmed platelet ageing was linked to a reduction decrease in cytoskeletal proteins, a reduction in mitochondria number, and lower calcium dynamics and granule secretion. This work delineates physical and functional changes in platelets as they age and serves as a base to examine differences associated with altered mean age of platelet populations in conditions such as immune thrombocytopenia and diabetes.