Impact of high platelet turnover on the platelet transcriptome: Results from platelet RNA-sequencing in patients with sepsis.

Thomas G Nührenberg,Björn A Grüning,Willibald Hochholzer,Christian Stratz,Vladimir Benes,Federico Marini,Marco Cederqvist,Jasmin Stöckle,Mark Zurek,Lutz Hein,Franz-Josef Neumann,Katherine James

doi:10.1371/journal.pone.0260222

Abstract

Sepsis is associated with high platelet turnover and elevated levels of immature platelets. Changes in the platelet transcriptome and the specific impact of immature platelets on the platelet transcriptome remain unclear. Thus, this study sought to address whether and how elevated levels of immature platelets affect the platelet transcriptome in patients with sepsis. Blood samples were obtained from patients with sepsis requiring vasopressor therapy (n = 8) and from a control group of patients with stable coronary artery disease and otherwise similar demographic characteristics (n = 8). Immature platelet fraction (IPF) was determined on a Sysmex XE 2100 analyser and platelet function was tested by impedance aggregometry. RNA from leukocyte-depleted platelets was used for transcriptome analysis by Next Generation Sequencing integrating the use of unique molecular identifiers. IPF (median [interquartile range]) was significantly elevated in sepsis patients (6.4 [5.3-8.7] % vs. 3.6 [2.6-4.6] %, p = 0.005). Platelet function testing revealed no differences in adenosine diphosphate- or thrombin receptor activating peptide-induced platelet aggregation between control and sepsis patients. Putative circular RNA transcripts were decreased in platelets from septic patients. Leukocyte contamination defined by CD45 abundance levels in RNA-sequencing was absent in both groups. Principal component analysis of transcripts showed only partial overlap of clustering with IPF levels. RNA sequencing showed up-regulation of 524 and down-regulation of 118 genes in platelets from sepsis patients compared to controls. Upregulated genes were mostly related to catabolic processes and protein translation. Comparison to published platelet transcriptomes showed a large overlap of changes observed in sepsis and COVID-19 but not with reticulated platelets from healthy donors. Patients with sepsis appear to have a less degraded platelet transcriptome as indicated by increased levels of immature platelets and decreased levels of putative circular RNA transcripts. The present data suggests that increased protein translation is a characteristic mechanism of systemic inflammation.

Highlights

Sepsis is the most common cause of death in hospitalised patients [1]
Immature platelet fraction (IPF) was determined on a Sysmex XE 2100 analyser and platelet function was tested by impedance aggregometry
Putative circular RNA transcripts were decreased in platelets from septic patients

Summary

Introduction

Sepsis is the most common cause of death in hospitalised patients [1]. Large amounts of circulating pathogens trigger a massive immune response and an activation of the coagulation system. Progressive dysfunction of the immune and coagulation system predisposes for multi-organ failure and death. Due to their homeostatic and immunomodulatory function, platelets play a key role in sepsis [2]. In addition to platelet activation, platelet turnover is increased in sepsis and levels of young, immature platelets may be associated with worse outcomes in this setting [7]. Sepsis as a state of increased platelet turnover may be seen as a model to study changes in platelet properties associated with higher platelet turnover. This study sought to address whether and how elevated levels of immature platelets affect the platelet transcriptome in patients with sepsis

Methods

Results

Conclusion