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 6

miR-483 reduces circulatory levels of LDL-C in mice.

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miR-483 reduces circulatory levels of LDL-C in mice. (A) Male and female...
(A) Male and female C57BL/6 mice were fed an HFD or chow diet for 6 weeks. A single dose of AAV8-null (AAV-null) or AAV8-pri-miR-483 (AAV-483) was administered by tail vein injection at the end of week 2. (B and C) Hepatic miR-483-5p levels were determined by qPCR, protein levels of PCSK9, LDLR, and CTGF were detected by Western blot. #CTGF in the same samples were detected in parallel in a separate gel (C). (D) Ago-1 was immunoprecipitated from fixed liver tissue, and Ago1-associated miR-483-5p, Pcsk9, and Ctgf mRNA levels were quantified by qPCR. (E) Total cholesterol levels measured by cholesterol assay. (F) Serum levels of VLDL, LDL, and HDL were determined by FPLC. (G) The correlations between hepatic miR-483-5p expression levels and serum levels of PCSK9 (left) or total cholesterol (right) are shown. ΔCT represents the difference between the cycle threshold of miR-483-5p and U6. The numbers of mice used are shown in Supplemental Table 1. Data are mean ± SEM. In B, non-normally distributed data were analyzed using Kruskal-Wallis test with Dunn’s multiple comparisons between indicated groups. In C and D, non-normally distributed data were analyzed using Mann-Whitney U test. In E, normally distributed data were analyzed by 1-way ANOVA test with a Bonferroni’s post hoc test between 2 indicated groups. In G, the correlation analysis was assessed by the Pearson method. *P < 0.05. miR, microRNA; LDL-C, LDL-cholesterol; HFD, high-fat diet; AAV8, adeno-associated virus 8; PCSK9, proprotein convertase subtilisin/kexin type 9; LDLR, LDL receptor; CTGF, connective tissue growth factor; VLDL, very LDL; IDL, intermediate-density lipoprotein.