Plasma from trauma patients induces transcriptional reprogramming of endothelial cell metabolism and inflammatory pathways

Introduction: Trauma causes a thromboinflammatory milieu characterized by coagulopathy and metabolic perturbations, however, the transcriptional changes of the endothelium following trauma remain unknown. We hypothesized that when exposed to severely injured ex-vivo trauma plasma, there would be induction of pro-inflammatory genes.

Materials & Methods: Citrated plasma was collected upon arrival from trauma patients at a level 1 trauma center. Patients were divided by injury severity score (ISS) and base deficit (BD) (Group 1 ISS < 15 BD < 6, Group 4 ISS ≥ 15 BD >6). HUVECs were grown to 80% confluence and incubated for 1 hour or 4 hours with 10% trauma patient ex-vivo plasma. We then performed a poly A-selected mRNA-seq analysis. A DESeq2 analysis was used for differential expression with Wald test and Benjamini-Hochberg correction to determine significance. Pathway analysis was performed using EnrichR via MSigDB Hallmark 2000 on those genes with p<0.05 and a log 2 fold change of less than -1 or greater than 1.

Results: The pathway analysis showed significantly different inflammatory and cell fate determination pathways between cells exposed to group 1 and group 4 plasma. The TNF alpha, NFkB, and TGF beta pathways were significantly downregulated in group 4 (Fig. 1a p<0.001). At the 4-hour time point, wnt signaling, which is involved in both cell fate determination, migration, and glucose homeostasis is downregulated in group 4 (Fig. 1b p<0.001).

Conclusion: Multiple pro-inflammatory signaling pathways are differentially expressed between cells exposed to group 1 versus group 4 plasma. At later timepoints, regulation of energy homeostasis and cell fate determination becomes predominant. The genes identified in this analysis may create novel therapeutic targets in the treatment of trauma patients.

Figure 1. RNA-seq pathway analysis at 1 and 4 hours

Figure 2. PPI map from the genes at the 4-hour time point analysis using OmicsNet with the InnateDB database

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