Shen Yahui, Wang Lina, Wu Yunhui, Ou Yingwei, Lu Huiyu, Yao Xin
Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
Department of Respiratory and Critical Care Medicine, Taizhou Clinical Medical School of Nanjing Medical University, Taizhou, Jiangsu 225300, P.R. China.
Exp Ther Med. 2021 Jul;22(1):717. doi: 10.3892/etm.2021.10149. Epub 2021 May 3.
Exosomal microRNAs (exo-miRNAs or miRs) have demonstrated diagnostic value in various diseases. However, their diagnostic value in chronic obstructive pulmonary disease (COPD) has yet to be fully established. The purpose of the present study was to screen differentially expressed exo-miRNAs in the plasma of patients with COPD and healthy individuals and to evaluate their potential diagnostic value in COPD. Differentially expressed exo-miRNAs in the plasma of patients with COPD and controls were identified using high-throughput sequencing and confirmed using reverse transcription-quantitative PCR (RT-qPCR). Bioinformatics analysis was then performed to predict the function of the selected exo-miRNAs and their target genes in COPD. After a network model was constructed, linear regression analysis was performed to determine the association between exo-miRNA expression and the clinical characteristics of subjects in a validated cohort (46 COPD cases; 34 matched healthy controls). Receiver operating characteristic curve was subsequently plotted to test the diagnostic value of the candidate biomarkers. The top 20 significantly aberrantly expressed COPD-associated exo-miRNAs were verified using RT-qPCR. Of these, nine were then selected for subsequent analysis, five of which were found to be upregulated (miR-23a, miR-1, miR-574, miR-152 and miR-221) and four of which were downregulated (miR-3158, miR-7706, miR-685 and miR-144). The results of Gene Ontology and KEGG pathway analysis revealed that these miRNAs were mainly involved in certain biological functions, such as metabolic processes, such as galactose metabolism and signaling pathways (PI3K-AKT) associated with COPD. The expression levels of three exo-miRNAs (miR-23a, miR-221 and miR-574) were found to be negatively associated with the forced expiratory volume in the 1st second/forced vital capacity. Furthermore, the area under the curve values of the three exo-miRNAs (miR-23a, miR-221 and miR-574) for COPD diagnosis were 0.776 [95% confidence interval (CI), 0.669-0.882], 0.688 (95% CI, 0.563-0.812) and 0.842 (95% CI, 0.752-0.931), respectively. In conclusion, the three circulating exosomal miRNAs (miR-23a, miR-221 and miR-574) may serve as novel circulating biomarkers for the diagnosis of COPD. These results may also enhance our understanding and provide novel potential treatment options for patients with COPD.
外泌体微小RNA（外泌体miRNA或miR）已在多种疾病中显示出诊断价值。然而，它们在慢性阻塞性肺疾病（COPD）中的诊断价值尚未完全确立。本研究的目的是筛选COPD患者和健康个体血浆中差异表达的外泌体miRNA，并评估其在COPD中的潜在诊断价值。使用高通量测序鉴定COPD患者和对照组血浆中差异表达的外泌体miRNA，并使用逆转录定量PCR（RT-qPCR）进行验证。然后进行生物信息学分析，以预测所选外泌体miRNA及其靶基因在COPD中的功能。构建网络模型后，进行线性回归分析以确定外泌体miRNA表达与验证队列（46例COPD病例；34例匹配的健康对照）中受试者临床特征之间的关联。随后绘制受试者工作特征曲线以测试候选生物标志物的诊断价值。使用RT-qPCR验证了前20个显著异常表达的COPD相关外泌体miRNA。其中，选择9个进行后续分析，发现其中5个上调（miR-23a、miR-1、miR-574、miR-152和miR-221），4个下调（miR-3158、miR-7706、miR-685和miR-144）。基因本体论和KEGG通路分析结果表明，这些miRNA主要参与某些生物学功能，如代谢过程，如半乳糖代谢以及与COPD相关的信号通路（PI3K-AKT）。发现三种外泌体miRNA（miR-23a、miR-221和miR-574）的表达水平与第1秒用力呼气量/用力肺活量呈负相关。此外，三种外泌体miRNA（miR-23a、miR-221和miR-574）用于COPD诊断的曲线下面积值分别为0.776 [95%置信区间（CI），0.669-（0.882）]、0.688（95%CI，0.563-0.812）和0.842（95%CI，0.752-0.931）。总之，三种循环外泌体miRNA（miR-23a、miR-221和miR-574）可能作为诊断COPD的新型循环生物标志物。这些结果也可能增进我们的理解，并为COPD患者提供新的潜在治疗选择。
