Effects of Human Bone Marrow Mesenchymal Stem Cell-Derived Extracellular Microvesicles on Non-diabetic and Diabetic Human Coronary Artery Endothelial Cells
Cynthia Xu, Catherine Karbasiafshar, Karla M. Braga, Rayane Brinck Teixeira, Frank Sellke, M. R. Abid
Cardiovascular Research Center, Pawtucket, Rhode Island, United States
Objective: Human bone marrow mesenchymal stem cell-derived extracellular vesicles (HBMSC-EV) have been shown in porcine models of chronic myocardial ischemia to improve angiogenesis and cardiac performance after intramyocardial injection, but have attenuated effects in metabolic syndrome. The purpose of this study was to determine the mechanisms responsible for the differential therapeutic effects of HBMSC-EV on non-diabetic and diabetic human coronary artery endothelial cells (HCAEC and DM-HCAEC), and to determine the mechanisms by which diabetes diminishes the therapeutic efficacy of HBMSC-EV. Design: HBMSC-EV were isolated from human bone marrow mesenchymal stem cells. HCAEC and DM-HCAEC were serum starved for 12 hours. Cells were then treated with either a vehicle or HBMSC-EV for 6 hours. After vehicle or HBMSC-EV treatment, a Reactive Oxygen Species (ROS) Assay was performed, and cell lysates were obtained for proteomics and western blot analysis. Setting: Basic science surgical laboratory. Patients: None. Interventions: Induction of oxidative stress, subsequent HBMSC-EV treatment. Main Outcome Measures: ROS levels, proteomic pathways related to proliferation, inflammation and cellular metabolism, western blot analysis. Results: ROS quantification showed higher level of ROS in DM-HCAEC compared to HCAEC, however, HBMSC-EV treatment resulted in significant decrease in ROS in both DM and non-DM HCAEC (Figure 1A). Gene ontology enrichment pathway analysis using the ShinyGO web application demonstrated up-regulation of RNA pathways (RNA processing, false discovery rate 3.9*10-18) and down-regulation of inflammatory pathways (Neutrophil activation and degranulation, false discovery rate 9.6*10-10 and 2.4*10-9) in HCAEC after HBMSC-EV treatment (Figure 1B). In DM-HCAEC, there was a diminished response to HBMSC-EV, likely due to basal differences between HCAEC and DM-HCAEC, with decreased expression of RNA pathways and increased inflammatory pathways in DM-HCAEC. Proteomics and western blot analyses demonstrated that DM-HCAEC had inappropriate MAPK activation in response to cellular starvation, which were corrected after HBMSC-EV treatment. Conclusions: These data demonstrate that HBMSC-EV had antioxidant and anti-inflammatory effects on coronary endothelial cells, but these benefits appear to be dampened in DM-HCAEC due to innate dysfunction. These findings underline the need for future efforts to augment the therapeutic potential of HBMSC-EV in patients with diabetes and cardiovascular disease.
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