PB0841 - Activated Protein C’s Cytoprotection Against Thromboinflammation: A Multi-Omic Study

Monday, June 24, 2024

13:45 – 14:45 ICT

Room: Exhibition Hall

Presenting Author(s)

OT

Otto N. Thielen, MD

General Surgery Resident
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Surgery
Aurora, Colorado, United States

Co-Author(s)

WH

William Hallas

General Surgery Resident
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Surgery
Aurora, Colorado, United States
SM

Sanchayita Mitra

Research Instructor
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Surgery
Aurora, Colorado, United States
FG

Fabia Gamboni

Research Instructor
University of Colorado
Aurora, Colorado, United States
JH

Julia Haines

Assistant Research Professor
University of Colorado
Aurora, Colorado, United States
CE

Christopher Erickson

Research Instructor
University of Colorado
Aurora, Colorado, United States
LG

Lauren T. Gallagher

General Surgery Resident
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Surgery
Aurora, Colorado, United States
BR

Benjamin J. Ramser

General Surgery Resident
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Surgery
Aurora, Colorado, United States
Benjamin W. Stocker, MD

General Surgery Resident
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Surgery
Denver, Colorado, United States
Ernest E. Moore, MD

Director of Research
Ernest E Moore Shock Trauma Center ay Denver Health
Denver, Colorado, United States
CS

Christopher C. Silliman

Professor
Vitalant Research Institute
Aurora, Colorado, United States
Laurent O. Mosnier

Professor
Scripps Research
La Jolla, California, United States
JG

John Griffin

Professor
The Scripps Research Institute
La Jolla, California, United States
KH

Kirk C. Hansen

Professor
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Biochemistry and Molecular Genetics
Aurora, Colorado, United States
AD

Angelo D'Alessandro

Professor
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Biochemistry and Molecular Genetics
Aurora, Colorado, United States
MC

Mitchell J. Cohen

Professor
University of Colorado Anschutz Medical Campus, School of Medicine, Department of Surgery
Aurora, Colorado, United States

Background: Activated Protein C (aPC) has been shown to drive both trauma-induced coagulopathy (TIC) by cleaving, and thus inactivating, factors Va and VIIIa and by re-depressing fibrinolysis while simultaneously mediating an inflammomodulatory milieu via protease-activated receptor 1 (PAR-1) cytoprotective signaling.

Aims: Here, we aimed to elucidate if treatment with 3K3A-aPC, a new variant with enhanced cytoprotection and minimal anticoagulant activity, would mitigate endothelial thromboinflammation and uncover important cytoprotective pathways and biomediators of aPC deranged by injury and shock.

Methods: Citrated plasma (TP) was collected upon arrival to a level 1 trauma center from patients in severe trauma and shock, defined as injury severity score (ISS) >15 and base deficit (BD) <-6. Primary endothelial cells (HUVECs) were incubated with 10% TP for 5 minutes (min), 30 min, and 1 hour (hr) with or without 30-min pre-incubation with 3K3A-aPC. Supernatant and cells were collected and analyzed with mass spectrometry multi-omic analysis, as previously described. Significant analytes were determined using false-discovery rate (FDR)-corrected 2-way analysis of variance (ANOVA, p< 0.05).

Results: Multi-omic results demonstrated responses to injury and shock after exposure for 1 hour. aPC mitigated shock-induced hyperuricemia via de-activation of the polyol pathway by restoring adenosine triphosphate (ATP) and urate levels. It maintained energy homeostasis and energy supply by stabilizing tricarboxylic acid (TCA) cycle, creatine phosphate shuttle, and glycolysis metabolites (Figure 1a). Finally, it stabilized numerous thromboinflammatory and barrier-protective proteins (Figure 1a and 1b).

Conclusion(s): The multi-omic findings of this study demonstrate trauma and shock-induced derangements in energy homeostasis involving the TCA cycle, creatine phosphate, and polyol pathways, as well as thromboinflammation via coagulation and complement pathways that are protected by aPC at the golden hour, when the effects of injury and shock start to manifest. This makes it an ideal therapeutic target for trauma patients that survive past their initial injury and resulting hemorrhagic shock.