The Impact of MiR-33a-5p Inhibition in Pro-Inflammatory Endothelial Cells.

• Type of Material: Article
• Authors: Kun Huang, Clemson University; Mark Pitman, Clemson University; Olanrewaju Oladosu, Clemson University; Jing Echesabal-Chen, Clemson University; Lucia Vojtech, University of Washington, Seattle; Ikechukwu Esobi, Clemson University; Jessica Larsen, Clemson University; Hanjoong Jo, Emory University; Alexis Stamatikos, Clemson University
• Language: English
• Date: 2023-06-24
• Publisher: MDPI
• Publication Version: Final Published Version
• Copyright Statement: © 2023 by the authors.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.3390/diseases11030088
• Title of Journal or Parent Work: Diseases
• Volume: 11
• Issue: 3
• Grant/Funding Information: This work was supported in part by the United States Department of Agriculture, National Institute of Food and Agriculture (USDA-NIFA), Hatch Project SC-1700577; Accession No. 1021291, and an NIH/NIGMS SC BioCRAFT Pilot Project Award (P30GM131959). Dr. Jing Echesabal-Chen was partially supported by a CU FELLOWS R-Initiatives grant bestowed by the Clemson University Division of Research.
• Keywords: vascular inflammation; endothelial dysfunction; endothelial activation; VCAM-1; nanoparticle; HDL; microRNA; reverse cholesterol transport; nanotherapy
• Research Categories: Engineering, Biomedical; Health Sciences, Nutrition; Biology, Molecular

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.