Proteomic Profile of Procoagulant Extracellular Vesicles Reflects Complement System Activation and Platelet Hyperreactivity of Patients with Severe COVID-19

Abstract

BackgroundExtracellular vesicles (EVs) are a valuable source of biomarkers and display the pathophysiological status of various diseases. In COVID-19, EVs have been explored in several studies for their ability to reflect molecular changes caused by SARS-CoV-2. Here we provide insights into the roles of EVs in pathological processes associated with the progression and severity of COVID-19.MethodsIn this study, we used a label-free shotgun proteomic approach to identify and quantify alterations in EV protein abundance in severe COVID-19 patients. We isolated plasma extracellular vesicles from healthy donors and patients with severe COVID-19 by size exclusion chromatography (SEC). Then, flow cytometry was performed to assess the origin of EVs and to investigate the presence of circulating procoagulant EVs in COVID-19 patients. A total protein extraction was performed, and samples were analyzed by nLC-MS/MS in a Q-Exactive HF-X. Finally, computational analysis was applied to signify biological processes related to disease pathogenesis.ResultsWe report significant changes in the proteome of EVs from patients with severe COVID-19. Flow cytometry experiments indicated an increase in total circulating EVs and with tissue factor (TF) dependent procoagulant activity. Differentially expressed proteins in the disease groups were associated with complement and coagulation cascades, platelet degranulation, and acute inflammatory response.ConclusionsThe proteomic data reinforce the changes in the proteome of extracellular vesicles from patients infected with SARS-CoV-2 and suggest a role for EVs in severe COVID-19.

Plasma tissue factor pathway inhibitor levels in coronavirus disease 2019 patients: a systematic review and meta-analysis

“…In addition, White et al [37] who investigated the TFPI values in platelet-poor plasma of noncritical and critical COVID-19 patients (as defined by WHO criteria), found that critical patients had significantly elevated TFPI compared with noncritical patients. In accord, Moraes et al ., who isolated plasma extracellular vesicles from healthy donors and patients with severe COVID-19 and employed a proteomic approach to discern variations in extracellular vesicle protein abundance, notably observed the exclusive presence of TFPI within the proteome of nonsurvivors [38]. In addition, Corey et al [35] also observed an association between the elevation of TFPI levels and the acute respiratory distress syndrome (ARDS) severity in COVID-19.…”

The Proteome of Extracellular Vesicles Released from Pulmonary Microvascular Endothelium Reveals Impact of Oxygen Conditions on Biotrauma

“…Besides TF, exosomes from COVID-19 patients have been shown by proteomic analysis to convey pro-inflammatory cargo that was effective in triggering inflammatory responses in cells of distant organs [45]. Generally, the proteome of EVs in COVID-19 shows abnormalities, such as complement activation and platelet hyperreactivity [46], and increased loads of vWF, UPAR, and ADAMTS13 that correlates with higher levels of the thrombotic marker, D-dimer, and increased disease severity. In our study, vWF was shown to be upregulated as an EV component under 0-21% O 2 but downregulated under 95% O 2 and it may also have an important impact on pro-coagulative activity of EVs.…”

Acute to post-acute COVID-19 thromboinflammation persistence: Mechanisms and potential consequences

“…Importantly, high concentrations of TF-bearing EVs in circulation correlate with disease severity and occurrence of thromboembolic events in severe COVID-19 patients, being proposed as a potential biomarker ( Guervilly et al, 2021 ). Platelets seem to be one of the main sources of TF-bearing circulating EVs during COVID-19, with severe patients presenting increased concentrations of platelet-derived EVs with higher surface expression of TF ( Moraes et al, 2022 ). Recently, we have reported the proteomic profile of circulating EVs from COVID-19 patients reflecting increased platelet degranulation, complement activation and fibrin clot formation ( Moraes et al, 2022 ).…”

“…Platelets seem to be one of the main sources of TF-bearing circulating EVs during COVID-19, with severe patients presenting increased concentrations of platelet-derived EVs with higher surface expression of TF ( Moraes et al, 2022 ). Recently, we have reported the proteomic profile of circulating EVs from COVID-19 patients reflecting increased platelet degranulation, complement activation and fibrin clot formation ( Moraes et al, 2022 ). A similar profile of circulating EVs was found to persist in individuals with PACS in our recent study.…”