Identification of a circulating microvesicle protein network involved in ST-elevation myocardial infarction

Abstract

Membrane microvesicles (MVs) are released from activated cells, most notably platelets, into the circulation. They represent an important mode of intercellular communication, and their number is increased in patients with acute coronary syndromes. We present here a differential proteomic analysis of plasma MVs from ST-elevation myocardial infarction (STEMI) patients and stable coronary artery disease (SCAD) controls. The objective was the identification of MVs biomarkers/drug targets that could be relevant for the pathogenesis of the acute event. Proteome analysis was based on 2D-DIGE, and mass spectrometry. Validations were by western blotting in an independent cohort of patients and healthy individuals. A systems biology approach was used to predict protein-protein interactions and their relation with disease. Following gel image analysis, we detected 117 protein features that varied between STEMI and SCAD groups (fold change cut-off ≥2; p<0.01). From those, 102 were successfully identified, corresponding to 25 open-reading frames (ORFs). Most of the proteins identified are involved in inflammatory response and cardiovascular disease, with 11 ORFs related to infarction. Among others, we report an up-regulation of α2-macroglobulin isoforms, fibrinogen, and viperin in MVs from STEMI patients. Interestingly, several of the proteins identified are involved in thrombogenesis (e.g. α2-macroglobulin, and fibrinogen). In conclusion, we provide a unique panel of proteins that vary between plasma MVs from STEMI and SCAD patients and that might constitute a promising source of biomarkers/drug targets for myocardial infarction.

Application of Extracellular Vesicles Proteomics to Cardiovascular Disease: Guidelines, Data Analysis, and Future Perspectives

“…Another example of these contaminants are lipoproteins, which are a major subcellular particle subset of plasma, thus it is not surprising that they are frequently found in EV preparations from plasma . For this reason, some groups performed protocols adding multiple washing steps (e.g., KBr or PBS) in order to avoid interference of plasma proteins even if this means a partial loss of EVs …”

“…The first proteomic studies of plasma‐derived EVs were in fact based on 2DE, and at that time were merely descriptive . An advantage of these 2DE‐based studies is the information regarding quick detection of proteoforms resulting from PTMs . This kind of information is not always obvious from a direct LC‐MS/MS analysis.…”

“…Indeed, during recent years, finding biomarkers in ACS patients has become an important issue in order to develop therapeutic strategies that may minimize myocardial injury. In this context, our group has recently performed the first high‐resolution 2D‐DIGE proteomic analysis of circulating MVs in plasma from STEMI patients compared to SCAD controls . A total of 117 proteoforms that varied between STEMI and SCAD groups were detected (fold change ≥ 2), from these 102 proteins were successfully identified by MS, corresponding to 25 unique proteins that were differentially regulated.…”

Low MT‐CO1 in Monocytes and Microvesicles Is Associated With Outcome in Patients With Coronary Artery Disease

“…Other studies showed that cardiovascular risk factors may change the content of circulating plasma microvesicles . A circulating microvesicle protein network involved in inflammatory response was found to be associated with STEMI . More than simply cell dust, extracellular vesicles are thus capable of transferring biological information to neighboring cells and play an active role in inflammatory diseases, including atherosclerosis and angiogenesis …”

Kalirin and CHD7: novel endothelial dysfunction indicators in circulating extracellular vesicles from hypertensive patients with albuminuria

“…EVs were isolated as previously described [22], with minor modifications. For more details see Supplementary material .…”