microRNA-483 ameliorates hypercholesterolemia by inhibiting PCSK9 production
Jianjie Dong, … , Sotirios Tsimikas, John Y.J. Shyy
Jianjie Dong, … , Sotirios Tsimikas, John Y.J. Shyy
Published October 29, 2020
Citation Information: JCI Insight. 2020;5(23):e143812. https://doi.org/10.1172/jci.insight.143812.
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Research Article Metabolism Vascular biology

microRNA-483 ameliorates hypercholesterolemia by inhibiting PCSK9 production

Abstract

Proprotein convertase subtilisin/kexin type 9 (PCSK9) affects cholesterol homeostasis by targeting hepatic LDL receptor (LDLR) for lysosomal degradation. Clinically, PCSK9 inhibitors effectively reduce LDL-cholesterol (LDL-C) levels and the incidence of cardiovascular events. Because microRNAs (miRs) are integral regulators of cholesterol homeostasis, we investigated the involvement of miR-483 in regulating LDL-C metabolism. Using in silico analysis, we predicted that miR-483-5p targets the 3′-UTR of PCSK9 mRNA. In HepG2 cells, miR-483-5p targeted the PCSK9 3′-UTR, leading to decreased PCSK9 protein and mRNA expression, increased LDLR expression, and enhanced LDL-C uptake. In hyperlipidemic mice and humans, serum levels of total cholesterol and LDL-C were inversely correlated with miR-483-5p levels. In mice, hepatic miR-483 overexpression increased LDLR levels by targeting Pcsk9, with a significant reduction in plasma total cholesterol and LDL-C levels. Mechanistically, the cholesterol-lowering effect of miR-483-5p was significant in mice receiving AAV8 PCSK9-3′-UTR but not Ldlr-knockout mice or mice receiving AAV8 PCSK9-3′-UTR (ΔBS) with the miR-483-5p targeting site deleted. Thus, exogenously administered miR-483 or similarly optimized compounds have potential to ameliorate hypercholesterolemia.

Authors

Jianjie Dong, Ming He, Jie Li, Ariane Pessentheiner, Chen Wang, Jin Zhang, Yameng Sun, Wei-Ting Wang, Yuqing Zhang, Junhui Liu, Shen-Chih Wang, Po-Hsun Huang, Philip L.S.M. Gordts, Zu-Yi Yuan, Sotirios Tsimikas, John Y.J. Shyy

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Figure 4

miR-483 overexpression in HepG2 cells increases LDL-C uptake.

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miR-483 overexpression in HepG2 cells increases LDL-C uptake. (A–H) HepG...
(A–H) HepG2 and mPCSK9 HepG2 cells were transfected with pre-483 or anti-483 as indicated. Fluorescent-labeled LDL was incubated with HepG2 and mPCSK9 HepG2 cells. LDL uptake was detected by flow cytometry (A and B) or confocal microscopy (C and D) (original magnification, ×20; scale bars: 10 μm). (E and F) Levels of PCSK9 in conditioned media were measured by ELISA and Western blot analysis. (G and H) HepG2 and mPCSK9 HepG2 cells were incubated with 1 μM atorvastatin for 24 hours. mRNA and protein levels of PCSK9 and LDLR were determined by qPCR and Western blot analysis. In MT HepG2 cells, #LDLR in the same samples were detected in parallel in a separate gel (H). Data are mean ± SEM from at least 4 independent experiments. In A, B, and F, non-normally distributed data were analyzed using Mann-Whitney U test between 2 groups. In C–E, normally distributed data were analyzed by 2-tailed Student’s t test with Welch correction between 2 groups. In G and H, non-normally distributed data were analyzed using Kruskal-Wallis test with Dunn’s multiple comparisons between indicated groups. *P < 0.05 vs. Ctrl or between 2 indicated groups. miR, microRNA; HepG2, human hepatocellular carcinoma; LDL-C, LDL-cholesterol; PCSK9, proprotein convertase subtilisin/kexin type 9.