Key result
A porcine model of acute myocardial infarction was used to evaluate relative fold changes of exosomal miR-1, miR-133, and miR-208 during a 90-minute ischemic period, with no quantitative results reported in the abstract.
Preclinical porcine AMI data remain inconclusive without quantitative results; leaves open exosomal miRNA biomarker potential.
Background: Circulating exosomes are small (40-100nm) extracellular vesicles secreted by various different cell types, such as endothelial cells, fibroblasts, tumor cells and lymphocytes and present in almost all biological fluids. They are important carriers of stable microRNAs (miRNAs) in plasma. miRNA-1 and miRNA-133 have previously been shown to rise several hours after acute myocardial infarction (AMI). miRNA-197 and miRNA-223 have been found to predict cardiovascular death in patients with symptomatic CAD. It has therefore been hypothesized that miRNAs may be used as biomarkers in the detection, diagnosis and monitoring of cardiac diseases and therapies. In this study we assessed the change of exosomal levels of miRNA-1, miRNA-133 and miRNA-208 during the ischemic period of AMI. Methods: In this study, we included 3 pigs which underwent a 90 minute period percutaneous occlusion of the mid left anterior descending artery (LAD) followed by reperfusion. EDTA blood samples were obtained at baseline and at 10, 30, 60 and 90 minutes of occlusion. Plasma was prepared for exosome isolation by progressive centrifugation steps of 1200 x g (10 minutes), 1800 x g (10minutes) and 10 000 x g (20 minutes) and then filtered through 0.2μm syringe filters. One mL of thusly prepared plasma was suspended in 9mL of PBS and ultra-centrifugated at 100 000 x g (120 minutes), followed by a washing step and another round of centrifugation at 100 000 x g (120 minutes). Nanoparticle Tracking and Western Blot of CD63 and CD9 confirmed the isolation of exosomes. miRNAs were isolated using QIAGEN miRNeasy Serum / Plasma kits, reverse transcribed using QIAGEN miScript RT kit and qPCR was performed using miScript SYBR® Green PCR Kit. Fold changes were normalized using ce-miR-39 Spike-in-Control of the QIAGEN Serum / Plasma kit. Relative fold changes of miR-1, miR-133 and miR-208 at baseline, 10, 30, 60 and 90 minutes after begin of reperfusion were evaluated.
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Mueller et al. (2018) studied acute myocardial infarction (n=3). 90 minute percutaneous occlusion of the mid LAD vs. baseline was evaluated on Relative fold changes of miR-1, miR-133 and miR-208. A porcine model of acute myocardial infarction was used to evaluate relative fold changes of exosomal miR-1, miR-133, and miR-208 during a 90-minute ischemic period, with no quantitative results reported in the abstract.
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