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 7

miR-483 targets Pcsk9 3′-UTR to reduce LDL-C levels in mice.

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miR-483 targets Pcsk9 3′-UTR to reduce LDL-C levels in mice. (A) Male an...
(A) Male and female C57BL/6 mice were administered AAV8-PCSK9-3′-UTR WT (WT) or AAV8-PCSK9-3′-UTR with a deleted miR-483 binding site (ΔBS) together with AAV-miR-483 or AAV-null by tail vein injection (n = 6–13 in each group). All mice were fed an HFD for 8–10 weeks. (B) Levels of Pcsk9 and Ldlr mRNA in mouse liver were determined by qPCR. (C) Protein levels of hepatic LDLR were detected by Western blot analysis. (D–F) Serum levels of total cholesterol, VLDL, LDL, and HDL are shown. (G) Serum levels of PCSK9 assessed by ELISA are shown. The numbers of mice used are shown in Supplemental Table 1. Data are mean ± SEM. In B, normally distributed data was analyzed by 1-way ANOVA test with a Bonferroni’s post hoc test between 2 indicated groups. In C, D, and G, non-normally distributed data were analyzed using Mann-Whitney U test between 2 indicated groups. *P < 0.05 vs. WT. miR, microRNA; LDL-C, LDL-cholesterol; AAV8, adeno-associated virus 8; PCSK9, proprotein convertase subtilisin/kexin type 9; AAV8-PCSK9-3′-UTR, AAV8-based recombinant virus encoding mouse Pcsk9 mRNA encompassing its native 3′-UTR; HFD, high-fat diet; LDLR, LDL receptor; VLDL, very LDL.