Abstract
Blood platelet RNA-sequencing is increasingly used among the scientific community. Aberrant platelet transcriptome is common in cancer or cardiovascular disease, but reference data on platelet RNA content in healthy individuals are scarce and merit complex investigation. We sought to explore the dynamics of platelet transcriptome. Datasets from 204 healthy donors were used for the analysis of splice variants, particularly with regard to age, sex, blood storage time, unit of collection or library size. Genes B2M, PPBP, TMSB4X, ACTB, FTL, CLU, PF4, F13A1, GNAS, SPARC, PTMA, TAGLN2, OAZ1 and OST4 demonstrated the highest expression in the analysed cohort, remaining substantial transcription consistency. CSF3R gene was found upregulated in males (fold change 2.10, FDR q < 0.05). Cohort dichotomisation according to the median age, showed upregulated KSR1 in the older donors (fold change 2.11, FDR q < 0.05). Unsupervised hierarchical clustering revealed two clusters which were irrespective of age, sex, storage time, collecting unit or library size. However, when donors are analysed globally (as vectors), sex, storage time, library size, the unit of blood collection as well as age impose a certain degree of between- and/or within-group variability. Healthy donor platelet transcriptome retains general consistency, with very few splice variants deviating from the landscape. Although multidimensional analysis reveals statistically significant variability between and within the analysed groups, biologically, these changes are minor and irrelevant while considering disease classification. Our work provides a reference for studies working both on healthy platelets and pathological conditions affecting platelet transcriptome.
Highlights
Blood platelet RNA-sequencing is increasingly used among the scientific community
The available literature on healthy platelet transcriptome mostly relies on a limited sample size[22,25,27,28]
In the presented study, which included over 200 samples, we have shown consistency of separately analysed splice reads which rarely surpassed criteria of statistical significance
Summary
Blood platelet RNA-sequencing is increasingly used among the scientific community. Aberrant platelet transcriptome is common in cancer or cardiovascular disease, but reference data on platelet RNA content in healthy individuals are scarce and merit complex investigation. Datasets from 204 healthy donors were used for the analysis of splice variants, with regard to age, sex, blood storage time, unit of collection or library size. When donors are analysed globally (as vectors), sex, storage time, library size, the unit of blood collection as well as age impose a certain degree of between- and/or within-group variability. The introduction of RNA sequencing (RNA-seq) provided even more detailed information on the number and biotypes of RNAs in platelets This method, combined with platelet sorting, allowed investigators to characterize and compare. The ultimate golden trove for platelet transcriptome analysis remains single platelet sequencing. This approach, is challenging due to the limited amount of input material, with one platelet estimated to have ~ 2.2 femtograms of total RNA mass[10]. Only single megakaryocyte RNA sequencing is feasible[11,12]
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call