Intra-Cardiac Release of Extracellular Vesicles Shapes Inflammation Following Myocardial Infarction

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  Figure 1.. Cardiac extracellular vesicles (EVs) are transiently released in response to myocardial infarction (MI) in mice and originate mainly from cardiomyocyte. A, Typical representative electronic microscopy of large EVs (lEVs). B, Representative size distribution analysis by tunable resistive pulse sensing (TRPS) of lEVs from ischemic and sham murine cardiac tissue 24 h postligation. C, lEVs release during 72 h after the onset of ischemia in sham (open circles) and MI mice (dark circles). D, Typical representative electronic microscopy of small EVs (sEVs). E, Representative size distribution analysis by TRPS of sEVs from ischemic and sham murine cardiac tissue 24 h postligation. F, sEVs release during 72 h after the onset of ischemia in shams (open circles) and MI mice (dark circles). G, Cellular origin of lEVs isolated from infarcted hearts 24 h postligation. H, Representative Western blots for Hsc70 and troponin T in ischemic left ventricle (LV) in lEV and sEVs derived from sham and MI. I, Representative Western blots for Hsc70, CD63, and CD9 in ischemic LV and sEVs. J, CD63 levels measured using integrated magnetic–electrochemical exosome and normalized to EV concentrations per…

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  Figure 2.. Cardiac extracellular vesicles (EVs) are engulfed by monocytes and shape the inflammatory response. A–C, Infiltration of neutrophils (Neutro; A), monocytes (Mono; B), and lymphocytes (Lympho; C) in sham (open circles) and myocardial infarction mice (dark circles) for 72 h after coronary artery ligation. Data are mean±SEM. n=4 to 7 animals per time point. **P<0.01 vs corresponding sham-operated mice at the same time point. D, Representative images of engulfment of GFP+ material by infiltrating immune cells (CD45+/CD11b+ leukocytes were gated on F4/80− monocytes and then further stratified by Ly6G and Ly6C expression) and (E) corresponding quantification. Data are mean±SEM. n=4 animals. *P<0.05 vs GFP+ cardiac neutrophils, ††P<0.01 vs GFP+ cardiac T lymphocytes. F–H, IL-6, CCL2, and CCL7 release by monocytes stimulated with cardiac-derived large (lEVs) or small EVs (sEVs). Data are mean±SEM. n=4 to 11 per experimental conditions. *P<0.05, **P<0.01 vs saline control.

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  Figure 3.. Intracardiac extracellular vesicles (EVs) in human myomectomy biopsies. A, Typical representative electronic microscopy of cardiac EVs from human biopsies indicating the presence of large (lEVs) and small EVs (sEVs). B, Representative size distribution analysis by tunable resistive pulse sensing (TRPS) of lEVs (in white) and sEVS (in grey). C, M-mode analysis of both lEVs and sEVs. D, EV concentration determined by TRPS. Data are mean±SEM. n=4 samples per fraction. *P<0.05 vs lEVs.