鉴定血浆 miR-4505、miR-4743-5p 和 miR-4750-3p 作为 2 型糖尿病患者冠心病的新型诊断生物标志物：病例对照研究。

Identification of plasma miR-4505, miR-4743-5p and miR-4750-3p as novel diagnostic biomarkers for coronary artery disease in patients with type 2 diabetes mellitus: a case-control study.

机构信息

Department of Endocrinology, Diabetology and Metabolic Diseases, Medical University of Lublin, Jaczewskiego 8, 20-090, Lublin, Poland.

Department of Clinical Genetics, Medical University of Lublin, Radziwillowska 11, 20-080, Lublin, Poland.

出版信息

Cardiovasc Diabetol. 2024 Jul 29;23(1):278. doi: 10.1186/s12933-024-02374-0.

DOI:10.1186/s12933-024-02374-0

PMID:39080630

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11287982/

Abstract

BACKGROUND

Type 2 diabetes mellitus (T2DM) and coronary artery disease (CAD) are commonly coexisting clinical entities with still growing incidence worldwide. Recently, circulating microRNAs (miRNAs) have emerged as novel molecular players in cardiometabolic diseases. This study aimed to identify a specific miRNA signature as a candidate biomarker for CAD in T2DM and to delineate potential miRNA-dependent mechanisms contributing to diabetic atherosclerosis.

METHODS

A total of 38 plasma samples from T2DM patients with and without CAD, CAD patients and healthy controls were collected for expression profiling of 2,578 miRNAs using microarrays. To investigate the regulatory role of differentially expressed (DE)-miRNA target genes, functional annotation and pathway enrichment analyses were performed utilizing multiple bioinformatics tools. Then, protein-protein interaction networks were established leveraging the STRING database in Cytoscape software, followed by cluster analysis and hub gene identification. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was carried out for microarray data validation in the larger replication cohort of 94 participants. Receiver operating characteristic analysis was applied to evaluate the diagnostic values of miRNAs. Multivariate logistic regression analysis was used to develop miRNA-based diagnostic models.

RESULTS

In the discovery stage, overexpression of hsa-miR-4505, hsa-miR-4743-5p, hsa-miR-6846-5p, and down-regulation of hsa-miR-3613-3p, hsa-miR-4668-5p, hsa-miR-4706, hsa-miR-6511b-5p, hsa-miR-6750-5p, hsa-miR-4750-3p, hsa-miR-320e, hsa-miR-4717-3p, hsa-miR-7850-5p were detected in T2DM-CAD patients. The DE-miRNA target genes were significantly enriched in calcium ion binding, regulation of actin cytoskeleton, and gene expression. hsa-miR-4505, hsa-miR-4743-5p, and hsa-miR-4750-3p were found to be involved in fatty acid metabolism, leukocyte transendothelial migration, and neurotrophin signaling pathway. Dysregulation of hsa-miR-4505, hsa-miR-4743-5p, and hsa-miR-4750-3p in T2DM-CAD patients compared with T2DM subjects and controls (all p < 0.001) was further confirmed by RT-qPCR. All validated miRNAs demonstrated good discriminatory values for T2DM-CAD (AUC = 0.833-0.876). The best performance in detecting CAD in T2DM was achieved for a combination of three miRNAs (AUC = 0.959, 100% sensitivity, 86.67% specificity).

CONCLUSIONS

Our study revealed a unique profile of plasma-derived miRNAs in T2DM patients with CAD. Potential miRNA-regulated pathways were also identified, exploring the underlying pathogenesis of CAD in T2DM. We developed a specific three-miRNA panel of hsa-miR-4505, hsa-miR-4743-5p and hsa-miR-4750-3p, that could serve as a novel non-invasive biomarker for CAD in patients with T2DM.

摘要

背景

2 型糖尿病（T2DM）和冠状动脉疾病（CAD）是常见的共存临床实体，在全球范围内发病率仍在不断上升。最近，循环 microRNAs（miRNAs）已成为心脏代谢疾病的新型分子标志物。本研究旨在确定一种特定的 miRNA 特征作为 T2DM 合并 CAD 的候选生物标志物，并阐明潜在的 miRNA 依赖机制导致糖尿病动脉粥样硬化。

方法

共收集了 38 例 T2DM 合并 CAD、CAD 患者和健康对照者的血浆样本，使用微阵列进行 2578 种 miRNA 的表达谱分析。为了研究差异表达（DE）-miRNA 靶基因的调控作用，利用多种生物信息学工具进行功能注释和通路富集分析。然后，利用 STRING 数据库在 Cytoscape 软件中建立蛋白质-蛋白质相互作用网络，随后进行聚类分析和枢纽基因识别。在更大的 94 名参与者的复制队列中，通过逆转录定量实时聚合酶链反应（RT-qPCR）对微阵列数据进行验证。应用受试者工作特征（ROC）分析评估 miRNA 的诊断价值。采用多元逻辑回归分析建立 miRNA 诊断模型。

结果

在发现阶段，T2DM-CAD 患者中 hsa-miR-4505、hsa-miR-4743-5p、hsa-miR-6846-5p 表达上调，hsa-miR-3613-3p、hsa-miR-4668-5p、hsa-miR-4706、hsa-miR-6750-5p、hsa-miR-4750-3p、hsa-miR-320e、hsa-miR-4717-3p、hsa-miR-7850-5p 表达下调。DE-miRNA 靶基因显著富集于钙离子结合、肌动蛋白细胞骨架调节和基因表达。hsa-miR-4505、hsa-miR-4743-5p 和 hsa-miR-4750-3p 参与脂肪酸代谢、白细胞跨内皮迁移和神经营养素信号通路。T2DM-CAD 患者与 T2DM 患者和对照组相比，hsa-miR-4505、hsa-miR-4743-5p 和 hsa-miR-4750-3p 的表达失调（均 p<0.001），进一步通过 RT-qPCR 得到验证。所有验证的 miRNA 对 T2DM-CAD 均具有良好的鉴别价值（AUC=0.833-0.876）。在检测 T2DM 合并 CAD 时，三种 miRNA 联合检测的效果最佳（AUC=0.959，敏感性 100%，特异性 86.67%）。

结论

本研究揭示了 T2DM 合并 CAD 患者血浆来源 miRNA 的独特特征。还确定了潜在的 miRNA 调节途径，探索了 T2DM 合并 CAD 的潜在发病机制。我们开发了一种特定的三 miRNA 组合（hsa-miR-4505、hsa-miR-4743-5p 和 hsa-miR-4750-3p），可作为 T2DM 患者 CAD 的新型非侵入性生物标志物。