载脂蛋白 B 代谢关键基因 PCSK9 的遗传（rs11591147）和表观遗传（DNA 甲基化）修饰与 CAD 患者 PCSK9 表达和血清蛋白的关系。

PCSK9 genetic (rs11591147) and epigenetic (DNA methylation) modifications associated with PCSK9 expression and serum proteins in CAD patients.

机构信息

Department of Genetics & Biotechnology, Osmania University, Hyderabad, Telangana State, India.

Vascular and Renal Translational Research Group, Institut de Recerca Biomèdica de Lleida (IRBLleida), Lleida, Spain.

出版信息

J Gene Med. 2021 Aug;23(8):e3346. doi: 10.1002/jgm.3346. Epub 2021 May 22.

DOI:10.1002/jgm.3346

PMID:33885177

Abstract

INTRODUCTION

Proprotein convertase subtilisin/kexin type 9 (PCSK9) genetic polymorphisms play a significant role in cholesterol homeostasis. Therefore, we aimed to investigate the association of PCSK9 genetic variations NM_174936.3:c.137G>T (R46L, rs11591147) and NM_174936.3:c.1120G>T (D374Y, rs137852912), as well as promoter DNA methylation status, with mRNA expression and circulating serum protein levels in coronary artery disease (CAD) patients.

METHODS

The present study includes 300 CAD cases and 300 controls from South India. Biochemical assays were performed using commercially available kits. PCSK9 rs11591147 and rs137852912 polymorphisms were analyzed by the polymerase chain reaction (PCR)-restriction fragment length polymorphism method, whereas promoter DNA methylation status and gene expression were determined using methylation specific PCR and quantitative PCR respectively.

RESULTS

The genotypic distribution of PCSK9 rs11591147 revealed that individuals with the TT-genotype and T-allele have a reduced risk for CAD. Furthermore, patients with the PCSK9 rs11591147 TT genotype have a significantly lower total cholesterol and low-density lipoprotein-cholesterol levels and also higher high-density lipoprotein-cholesterol levels than individuals with the GG genotype. Logistic regression analysis has shown that the GG and GT (p = 1.51 × 10 , p = 1.47 × 10 ) genotypes predicted the risk for CAD with an odds ratio of 5.8 and 7.3 respectively. In addition, individuals with the TT genotype were hypermethylated at promoter DNA of PCSK9, resulting in lower mRNA expression and circulating serum proteins than in individuals with the GG genotype. In silico analyses revealed that rs11591147 T-allele has protein destabilizing capacity.

CONCLUSIONS

In conclusion, the present study indicates that the PCSK9 gene expression and circulating serum protein levels are not only associated with rs11591147 genotype, but also with promoter DNA methylation. Furthermore, the findings with respect to both single nucleotide polymorphism and promoter DNA methylation may open avenues for novel treatment possibilities targeting PCSK9 for CAD management.

摘要

简介

前蛋白转化酶枯草溶菌素/ kexin 9 型（PCSK9）基因多态性在胆固醇稳态中起着重要作用。因此，我们旨在研究 PCSK9 基因变异 NM_174936.3:c.137G>T（R46L，rs11591147）和 NM_174936.3:c.1120G>T（D374Y，rs137852912）以及启动子 DNA 甲基化状态与冠状动脉疾病（CAD）患者 mRNA 表达和循环血清蛋白水平之间的关联。

方法

本研究纳入了来自印度南部的 300 例 CAD 患者和 300 例对照。使用商业试剂盒进行生化检测。通过聚合酶链反应（PCR）-限制性片段长度多态性方法分析 PCSK9 rs11591147 和 rs137852912 多态性，而启动子 DNA 甲基化状态和基因表达则分别通过甲基化特异性 PCR 和定量 PCR 来确定。

结果

PCSK9 rs11591147 的基因型分布表明，TT 基因型和 T 等位基因的个体患 CAD 的风险降低。此外，与 GG 基因型相比，PCSK9 rs11591147 TT 基因型的患者总胆固醇和低密度脂蛋白胆固醇水平明显降低，而高密度脂蛋白胆固醇水平明显升高。Logistic 回归分析显示，GG 和 GT（p=1.51×10-5，p=1.47×10-5）基因型的个体患 CAD 的风险分别为 5.8 和 7.3 倍。此外，TT 基因型的个体在 PCSK9 启动子 DNA 中发生高度甲基化，导致其 mRNA 表达和循环血清蛋白水平低于 GG 基因型的个体。计算机分析表明，rs11591147 T 等位基因具有蛋白不稳定性。