FIGURE 2.

Heparanase (HPSE) mRNA expression and ribosomal protected RNAs are significantly and dynamically upregulated in platelets during clinical sepsis. A, Quantitative real-time polymerase chain reaction (qRT-PCR) for HPSE mRNA was performed in platelets isolated from an independent cohort of septic patients or matched healthy donors. The fold change in HPSE expression in septic patients, compared to the average HPSE expression in healthy donors (denoted with a dotted line) was plotted. Statistical significance was measured using an unpaired non-parametric Mann-Whitney statistical analysis (*≤.05; average HPSE expression in healthy subjects is shown with a dotted line). B, RNA-seq analysis was performed on platelets isolated from the septic patients assessed within 72 h of intensive care unit admission for sepsis (Day 1) and then, in the same patients, approximately 90 days later during recovery. Statistical significance was measured using a one-way analysis of variance (***≤.0001; average HPSE expression in healthy subjects is shown with a dotted line). C, Ribosomal protected regions (RPR), an indicator of RNAs undergoing active translation, were increased in platelets from septic patients. The fold-change of HPSE RPR (measured in fragments per kilobase of transcript per million mapped reads [FPKM]) in septic patients, related to average levels in healthy donors (denoted with a dotted line) was plotted. An adjusted P-value was used to determine statistical significance (****<.0001). D, Periodicity of HPSE was used to determine where the ribosome was present on the codon. 1st represents the first nucleic acid in a codon, 2nd the second, and 3rd the third (*<0.05)