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Published on: September 15, 2017
The Impact of MiR-33a-5p Inhibition in Pro-Inflammatory Endothelial Cells
Kun Huang1, Mark Pitman2, Olanrewaju Oladosu1
1Department of Food, Nutrition, and Packaging Sciences, Clemson University, Clemson, SC 29634, USA.
Insights
Inhibiting miR-33a-5p in pro-inflammatory endothelial cells increases cholesterol efflux and shows atheroprotective effects. Targeted delivery of anti-miR-33a-5p offers a promising anti-atherogenic strategy.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Biotechnology
Background:
- Cholesterol accumulation in pro-inflammatory endothelial cells (EC) is implicated in atherogenesis.
- MicroRNA-33a-5p (miR-33a-5p) is a potential therapeutic target for atherosclerosis.
- Upregulating ABCA1 enhances cholesterol efflux, a key factor in preventing atherosclerosis.
Purpose of the Study:
- To investigate if inhibiting miR-33a-5p in pro-inflammatory EC can enhance ABCA1-dependent cholesterol efflux.
- To evaluate the efficacy of polymersome-mediated delivery of anti-miR-33a-5p to inflamed EC.
- To assess the atheroprotective potential of targeting miR-33a-5p in endothelial cells.
Main Methods:
- Transfection of LPS-challenged mouse aortic EC (iMAEC) with pAntimiR33a5p or control plasmid.
- Utilizing polymersomes for targeted delivery of anti-miR-33a-5p to inflamed iMAEC.
- Measuring ABCA1 protein expression and apoAI-mediated cholesterol efflux.
Main Results:
- Transfection with pAntimiR33a5p significantly increased ABCA1 protein and cholesterol efflux in iMAEC.
- Polymersomes selectively targeted pro-inflammatory iMAEC.
- Polymersome-delivered anti-miR-33a-5p significantly reduced miR-33a-5p expression in iMAEC.
Conclusions:
- Inhibition of miR-33a-5p in pro-inflammatory EC demonstrates atheroprotective effects.
- Targeted delivery of anti-miR-33a-5p via polymersomes is a viable anti-atherogenic strategy.
- This approach holds promise for preventing and treating atherosclerosis.
Abstract:
Evidence suggests cholesterol accumulation in pro-inflammatory endothelial cells (EC) contributes to triggering atherogenesis and driving atherosclerosis progression. Therefore, inhibiting miR-33a-5p within inflamed endothelium may prevent and treat atherosclerosis by enhancing apoAI-mediated cholesterol efflux by upregulating ABCA1. However, it is not entirely elucidated whether inhibition of miR-33a-5p in pro-inflammatory EC is capable of increasing ABCA1-dependent cholesterol efflux. In our study, we initially transfected LPS-challenged, immortalized mouse aortic EC (iMAEC) with either pAntimiR33a5p plasmid DNA or the control plasmid, pScr. We detected significant increases in both ABCA1 protein expression and apoAI-mediated cholesterol efflux in iMAEC transfected with pAntimiR33a5p when compared to iMAEC transfected with pScr. We subsequently used polymersomes targeting inflamed endothelium to deliver either pAntimiR33a5p or pScr to cultured iMAEC and showed that the polymersomes were selective in targeting pro-inflammatory iMAEC. Moreover, when we exposed LPS-challenged iMAEC to these polymersomes, we observed a significant decrease in miR-33a-5p expression in iMAEC incubated with polymersomes containing pAntimR33a5p versus control iMAEC. We also detected non-significant increases in both ABCA1 protein and apoAI-mediated cholesterol in iMAEC exposed to polymersomes containing pAntimR33a5p when compared to control iMAEC. Based on our results, inhibiting miR-33a-5p in pro-inflammatory EC exhibits atheroprotective effects, and so precisely delivering anti-miR-33a-5p to these cells is a promising anti-atherogenic strategy.
