Coagulation potential and the integrated omics of extracellular vesicles from COVID-19 positive patient plasma

Extracellular vesicles (EVs) participate in cell-to-cell communication and contribute toward homeostasis under physiological conditions. But EVs can also contribute toward a wide array of pathophysiology like cancer, sepsis, sickle cell disease, and thrombotic disorders. COVID-19 infected patients are at an increased risk of aberrant coagulation, consistent with elevated circulating levels of ultra-high molecular weight VWF multimers, D-dimer and procoagulant EVs. The role of EVs in COVID-19 related hemostasis may depend on cells of origin, vesicular cargo and size, however this is not well defined.

Researchers at the University of Maryland School of Medicine hypothesized that the procoagulant potential of EV isolates from COVID-19 (+) patient plasmas could be defined by thrombin generation assays. Here the researchers isolated small EVs (SEVs) and large EVs (LEVs) from hospitalized COVID-19 (+) patient (n = 21) and healthy donor (n = 20) plasmas. EVs were characterized by flow cytometry, Transmission electron microscopy, nanoparticle tracking analysis, plasma thrombin generation and a multi-omics approach to define coagulation potential. These data were consistent with differences in EV metabolite, lipid, and protein content when compared to healthy donor plasma isolated SEVs and LEVs. Taken together, the effect of EVs on plasma procoagulant potential as defined by thrombin generation and supported by multi-omics is enhanced in COVID-19. Further, the researchers observe that this effect is driven both by EV size and phosphatidyl serine.

Characterization of EVs

(A,B) particle size vs. percent intensity graphs of healthy donor (A) and COVID-19 (+) patients (B) SEVs/LEVs. (C) Mean of n = 5 sample size graph is mentioned. (D) TEM images of healthy donor and COVID-19 (+) patients SEVs and LEVs (representative image). An n = 5 each of healthy and COVID-19 SEVs and LEVs was used for particle size and TEM analysis. (E) Western blotting analysis of EV specific tetraspanin markers (CD63 and CD81) of healthy donor SEVs/LEVs and COVID-19 (+) SEVs/LEVs and EDP as negative control.