Abstract
Extracellular vesicles (EVs) are submicron, membrane-enclosed particles that are released from cells in various pathophysiological states. The molecular cargo of these vesicles is considered to reflect the composition of the cell of origin, and the EV proteome is therefore a potential source of biomarkers for various diseases. Our aim was to determine whether EVs isolated from plasma provide additional diagnostic value or improved pathophysiological understanding compared to plasma alone in the context of myocardial infarction (MI). A panel of proximity extension assays (n = 92) was employed to analyze EV lysates and plasma from patients with MI (n = 60) and healthy controls (n = 22). After adjustment for multiple comparisons, a total of 11 dysregulated proteins were identified in EVs of MI patients compared to the controls (q < 0.01). Three of these proteins: chymotrypsin C (CTRC), proto-oncogene tyrosine-protein kinase SRC (SRC) and C-C motif chemokine ligand 17 (CCL17) were unaltered in the corresponding plasma samples. As biomarkers for MI, rudimentary to no evidence exists for these proteins. In a separate group of patients with varying degrees of coronary artery disease, the decrease in EV-associated (but not plasma-related) SRC levels was confirmed by ELISA. Confirmation of the presence of SRC on EVs of different sizes and cellular origins was performed with ELISA, flow cytometry and nanoparticle tracking analysis. In conclusion, the data revealed that despite a similarity in the EV and plasma proteomes, analysis of isolated EVs does indeed provide additional diagnostic information that cannot be obtained from plasma alone.
Highlights
Extracellular vesicles (EV) are a heterogeneous group of membrane-limited particles released from cells upon stress, activation or injury[1]
We re-analyzed a previously published data set[27] comprising a total of 30 acoustically trapped and 30 centrifuged EV samples from 10 healthy donors assayed with the Olink cardiovascular disease (CVD) II proximity extension assays (PEA) Panel. 59 and 55 proteins were detected in centrifuged and trapped EV samples, respectively
These results demonstrate that EV isolation technique has very limited effect on the proteomic profile measured by PEA
Summary
Extracellular vesicles (EV) are a heterogeneous group of membrane-limited particles released from cells upon stress, activation or injury[1]. Endothelial cell-derived microvesicles[13] and Velez et al surveyed the proteome of plasma vesicles from patients with ST-elevation myocardial infarction (STEMI) patients and stable coronary artery disease[14]. We have previously compared the proteomic profiles of plasma EV from patients after an MI with those from healthy controls[15]. These studies all utilized mass spectrometry-based approaches that are known under such circumstances to have limited sensitivity and a bias towards abundant proteins[16]. Plasma EVs were isolated from MI patients and healthy controls with acoustic seed trapping[19] and the levels of > 90 low-abundance proteins were analyzed with proximity extension assays (PEA). Our aim was to compare the proteomic profiles of EVs and plasma, identify biomarker candidates that were unique to EVs, evaluate the diagnostic accuracy of these biomarkers and characterize these with respect to size and cellular origin
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