High density lipoprotein from coronary artery disease patients caused abnormal expression of long non-coding RNAs in vascular endothelial cells

doi:10.1016/j.bbrc.2017.04.082

Biochemical and Biophysical Research Communications

Volume 487, Issue 3, 3 June 2017, Pages 552-559

https://doi.org/10.1016/j.bbrc.2017.04.082 Get rights and content

Highlights

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Normal and aberrant HDL cause different expression of lncRNAs, genes, and mRNAs.
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There exist complex interactions among lncRNAs, encoding genes and miRNAs.
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The abnormal expression of lncRNAs involved in the regulation of vascular function.

Abstract

Increased evidence has showed that normal high density lipoprotein (HDL) could convert to dysfunctional HDL in diseases states including coronary artery disease (CAD), which regulated vascular endothelial cell function differently. Long non-coding RNAs (lncRNAs) play an extensive role in various important biological processes including endothelial cell function. However, whether lncRNAs are involved in the regulation of HDL metabolism and HDL-induced changes of vascular endothelial function remains unclear. Cultured human umbilical vein endothelial cells (HUVECs) were treated with HDL from healthy subjects and patients with CAD and hypercholesterolemia for 24 h, then the cells were collected for lncRNA-Seq and the expressions of lncRNAs, genes and mRNAs were identified. The bioinformatic analysis was used to evaluate the relationship among lncRNAs, encoding genes and miRNAs. HDL from healthy subjects and patients with CAD and hypercholesterolemia leaded to different expressions of lncRNAs, genes and mRNAs, and further analysis suggested that the differentially expressed lncRNAs played an important role in the regulation of vascular endothelial function. Thus, HDL from patients with CAD and hypercholesterolemia could cause abnormal expression of lncRNAs in vascular endothelial cells to affect vascular function.

Introduction

High density lipoprotein (HDL) is a large complex molecule which contains a variety of proteins, lipids and miRNAs [1], [2], [3]. HDL has been thought to exert protection against cardiovascular disease (CVD) by regulating cholesterol efflux from peripheral tissues, anti-inflammatory and antioxidant, nitric oxide (NO)–promoting function and promoting angiogenesis [4], [5], [6], [7], [8]. Despite epidemiological study had demonstrated that HDL level was inversely associated with cardiovascular risk [9], however, the failures of the clinical trials that designed to raise HDL-C level to reduce the cardiovascular events illustrated that the HDL biological activities or function were more important than its plasma level in diseased conditions [10], [11], [12]. In addition, the studies of genetic variants did not find a positive correlation between the plasma HDL level and coronary artery disease (CAD) [13]. Increased evidence has showed that normal HDL could convert to dysfunctional HDL in diseases states, which regulated vascular endothelial cell function differently [6], [14], [15]. As a result, the efforts to increase plasma HDL level have been transferred to the study and improvement of HDL function.

Long non-coding RNAs (lncRNAs) are a large class of RNAs with length longer than 200 nucleotides, exhibiting alternative spliced and polyadenylated, and many are poorly conserved and species specific [16], [17]. LncRNAs play an extensive role in various important biological processes such as chromatin modification [18], transcriptional and translational regulation [19], and many human diseases [20], [21]. The study of lncRNAs in cardiovascular system is still preliminary. LncRNAs have been identified in cardiomyocytes and vascular endothelial cells and were involved in cardiovascular development and diseases [22], [23], [24]. There are only a few studies investigating the associations between lncRNAs and cholesterol metabolism. Recently, it was reported that a long non-coding natural antisense transcript, APOA1-AS, could negatively regulate the expression of APOA1 through modulating the histone methylation patterns on the chromatin regions flanking the APOA1 gene [25]. Another study has uncovered the interaction between lncRNA and miRNA, in which the RP5-833A20.1 could inhibit the expression of nuclear factor I a (NFIA) by promoting the miR-382-5p expression, ultimately contributing to the regulation of cholesterol homeostasis, inflammatory responses, and foam cell formation [26].

These above studies have suggested a critical role for lncRNAs in the regulation of HDL metabolism and HDL-induced changes of vascular endothelial function. But the details remain unclear. In the present study, lncRNA-sequencing was applied for screening the genome-wide changes of lncRNAs in the cultured human umbilical vein endothelial cells (HUVECs) treated with healthy subjects HDL (n-HDL) and HDL from patients with CAD and hypercholesterolemia (p-HDL). The differential expressions of lncRNAs, genes and mRNAs were examined, and then the relationships among them and their relations with vascular endothelial function were analyzed.

Section snippets

Study population

Patients with CAD and hypercholesterolemia were enrolled. CAD were documented by angiography with the diagnostic criteria of ≥70% stenosis of at least one major coronary vessel caused by atherosclerosis. Patients with valvular heart diseases, congenital heart diseases, diabetes, infectious diseases, severe traumas and receiving operations in early last month were excluded. Healthy adult volunteers were selected as normal controls. All subjects were above 18 years old. This study was approved by

General clinical data of the two groups and their HDL inflammatory index

Except for lipid profile and medications, other characteristics were similar between healthy subjects and patients (Table 1). The proinflammatory index of HDL was evaluated by relative fluorescence units (RFU) and proinflammatory index of HDL in patients with CAD and hypercholesterolemia (RFU: 1821.73 ± 55.44) was much higher than that in healthy subjects (RFU: 888.79 ± 29.29).

RNA-Seq mapped reads, coverage and distribution in the genome

There were more than 100 million of reads obtained in each sample, the number of reads and mapping results were showed (

Discussion

The present study for the first time to our knowledge revealed the molecular events on a genome-wide scale that orchestrated the functions of vascular endothelial cells exposed to HDL from healthy subjects and CAD patients. Our RNA-Seq data revealed that the profiles of lncRNAs, genes and mRNAs were obviously different between n-HDL and p-HDL treated HUVECs. Moreover, KEGG analysis showed that the biological pathways of n-HDL treated cells were quite distinct from that in p-HDL group, with

Conflict of interest

The authors declare that they have no conflict of interest.

Sources of funding

This study was supported by the National Natural Science Foundation of China (81170271, 81370370, 81600382, 81670392, 81570213, 91439125, 81570329 and Distinguished Young Scholar 81325001), the Changjiang Scholars Program from Ministry of Education of China (2014), the Guangdong Pearl River Scholars Program (2012), International cooperation project (2015DFA31070) and National Major Research Program (2016YFC0903000) from the Ministry of Science and Technology of China, Guangdong Natural Science

Acknowledgments

We thank the patients and staffs at the First Affiliated Hospital, Sun Yat-sen University for their assistance throughout this study.

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¹: These two authors contributed equally to this study.

²: Both Zhi-Jun Ou and Jing-Song Ou contributed equally to the design and supervision of the research for this article.

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