Available online 16 April 2024, 117554

Author links open overlay panel, , , , , , , , Highlights•

LEPIS promotes the occurrence and development of atherosclerosis.

•

LEPIS regulates liver cholesterol metabolism and blood lipid levels by targeting TMOD4.

•

LEPIS enhances the stability of TMOD4 mRNA by promoting the nucleocytoplasmic shuttle of HuR.

Abstract:Background and aims

Long noncoding RNAs (lncRNAs) play important roles throughout the progression of atherosclerosis. In this study, we identified an uncharacterized lncRNA, Liver Expressions by PSRC1 Induced Specifically (LEPIS). This study aimed to clarify the mechanism though which LEPIS affects atherosclerosis (AS).

Methods

The expression of LEPIS and its potential target, tropomodulin 4 (TMOD4), was increased in the livers of ApoE-/- mice fed a high-fat diet (HFD). An ApoE-/- mouse model in which LEPIS or TMOD4 was overexpressed in the liver was established. The plaque load in the aorta was assessed, plasma was collected to measure blood lipid levels, and the liver was collected to study cholesterol metabolism.

Results

We found that both LEPIS and TMOD4 increased the AS burden and reduced hepatic cholesterol levels. Further study revealed that LEPIS and TMOD4 affected the expression of genes related to hepatic cholesterol homeostasis, including proprotein convertase subtilisin/kexin type 9 (PCSK9) and low-density lipoprotein receptor (LDLR), which are closely related to hypercholesterolemia. Mechanistically, human antigen R (HuR), an RNA-binding protein (RBP), was shown to be critical for the regulation of TMOD4 by LEPIS. Furthermore, we found that the overexpression of LEPIS promoted the shuttling of HuR from the nucleus to the cytoplasm, enhanced the stability of TMOD4 mRNA, and in turn promoted the expression of TMOD4. In addition, TMOD4 was found to affect intracellular cholesterol levels through PCSK9.

Conclusions

These results suggest that the LEPIS-HuR-TMOD4 axis is a potential intervention target for dysregulated hepatic cholesterol homeostasis and AS and may provide the basis for further reductions in the circulating LDL-C concentration and arterial plaque burden.

Section snippetsINTRODUCTION

Cardiovascular disease (CVD) is the most important chronic noncommunicable disease threatening human life and health worldwide [1]. Atherosclerotic cardiovascular diseases (ASCVDs), such as ischemic heart disease and ischemic stroke, are an important component of CVD [2]. In China, ASCVD is the top cause of death among urban and rural residents and accounts for more than 40% of total deaths [3]. In recent years, the ASCVD burden in China has continued to increase, and the prevention and control

Adeno-associated virus construction and effect verification.

LEPIS cDNA and TMOD4 cDNA were amplified via PCR and subsequently cloned and inserted into the pAV-CMV Globin Intron-MCS-P2A-EGFP-SV40polyA vector, after which the correct sequences of the LEPIS and TMOD4 genes in this construct were verified by sequencing. These constructs (referred to as AAV-LEPIS and AAV-TMOD4, respectively) and the corresponding vector (AAV-NC) were used to transfect mice. Similarly, the resulting recombinant plasmid was obtained (referred to as siTMOD4). The efficient

LEPIS and its potential target TMOD4 are involved in cholesterol homeostasis and AS.

ENSMUST00000125548, which is located on chromosome 3 near the coding gene TMOD4 and has six exons (Supplementary Figs. 1A and B), was the most significantly differentially expressed gene in the liver tissue between PSRC1-overexpressing ApoE-/- mice and ApoE-/- mice [9]. Based on an online prediction of coding probability, LEPIS had weak protein-coding ability (Supplementary Fig. 1C). Conservation analysis suggested that the exons are highly conserved across different mammalian species

DISCUSSION

Our study shows that LEPIS in the liver can induce the transfer of HuR from the nucleus to the cytoplasm, thereby increasing the stability of TMOD4 mRNA, which in turn enhances the expression of TMOD4. TMOD4 binds to the promoter of PCSK9, and enhances its transcription leading to elevated expression of PCSK9. This leads to a decrease in LDLR expression of hepatocytes, resulting in impaired cholesterol uptake by hepatocytes, leading to an increase in plasma LDL-Cl levels, which ultimately

Financial support

This work was supported by the Youth Fund of National Natural Science Foundation of China (81900398), the National Natural Science Foundation of China (82170474), the National Natural Science Foundation of China (82170274), Guangdong Basic and Applied Basic Research Foundation (2019A1515010666), Clinical Research Startup Program of Southern Medical University by High Level Construction Funding of Guangdong provincial Department of Education (LC2016PY002), and Clinical Research Program of

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Declaration of Competing Interest

☐ √ The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

☐The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:

REFERENCES (33)D. Zhao et al.Epidemiology of cardiovascular disease in China: current features and implications

Nature reviews. Cardiology

(2019)

K. Guo et al.PSRC1 overexpression attenuates atherosclerosis progression in apoE(-/- ) mice by modulating cholesterol transportation and inflammation

J MOL CELL CARDIOL

(2018)

R. Liu et al.Human antigen R: A potential therapeutic target for liver diseases

PHARMACOL RES

(2020)

R.P. Giugliano et al.Clinical efficacy and safety of achieving very low LDL-cholesterol concentrations with the PCSK9 inhibitor evolocumab: a prespecified secondary analysis of the FOURIER trial

Lancet (London, England)

(2017)

Y. Hu et al.A lincRNA-DYNLRB2-2/GPR119/GLP-1R/ABCA1-dependent signal transduction pathway is essential for the regulation of cholesterol homeostasis

J LIPID RES

(2014)

X. Zhao et al.The switch role of the Tmod4 in the regulation of balanced development between myogenesis and adipogenesis

GENE

(2013)

N. Townsend et al.Epidemiology of cardiovascular disease in Europe

Nature reviews. Cardiology

(2022)

S. Yusuf, S. Hawken, S. Ounpuu, T. Dans, A. Avezum, F. Lanas, M. McQueen, A. Budaj, P. Pais, J. Varigos, L. Lisheng,...L. Tokgözoğlu et al.The dawn of a new era of targeted lipid-lowering therapies

EUR HEART J

(2022)

B.A. Ference et al.Low-density lipoproteins cause atherosclerotic cardiovascular disease. 1. Evidence from genetic, epidemiologic, and clinical studies. A consensus statement from the European Atherosclerosis Society Consensus Panel

EUR HEART J

(2017)

E.S. Ford et al.Explaining the decrease in U.S. deaths from coronary disease, 1980-2000

The New England journal of medicine

(2007)

Q. Meng et al.Regulatory Roles of Related Long Non-coding RNAs in the Process of Atherosclerosis

FRONT PHYSIOL

(2020)

M. Wei et al.The impact of PSRC1 overexpression on gene and transcript expression profiling in the livers of ApoE(-/- ) mice fed a high-fat diet

MOL CELL BIOCHEM

(2020)

S. Srikantan et al.HuR function in disease

Frontiers in bioscience (Landmark edition)

(2012)

C.W. Schultz et al.Understanding and targeting the disease-related RNA binding protein human antigen R (HuR)

Wiley interdisciplinary reviews. RNA

(2020)

J. Luo et al.Mechanisms and regulation of cholesterol homeostasis

Nature reviews. Molecular cell biology

(2020)

View full text

© 2024 Elsevier B.V. All rights reserved.