Table_1_Serum MyomiRs as Biomarkers for Female Carriers of Duchenne/Becker Muscular Dystrophy.docx

Background: Duchenne/Becker muscular dystrophy (DMD/BMD) is an X-linked recessive lethal neuromuscular disease. MicroRNAs expressed in striated muscle, myomiRs, have been proposed as its potential biomarkers. Serum creatine kinase (CK) is commonly used as a biomarker in clinical practice, but it is not reliable. The aim of this study was to assess whether serum levels of myomiRs has diagnostic value for detection of female DMD/BMD carriers with normal or elevated CK.Methods: Thirty four female carriers and 33 age-matched healthy female controls were enrolled. Peripheral blood samples were collected and serum miRNAs were extracted for measurement of miR-1, miR-133a, miR-133b, miR-206, miR-208a, miR-208b, and miR-499 by quantitative real-time polymerase chain reaction.Results: MiR-1, miR-133a, miR-133b, miR-206, miR-208a, miR-208b, and miR-499 were upregulated in all female carriers in comparison to healthy controls. MiR-1 (Spearman's rho = +0.406, p = 0.017) was correlated with CK in the female carrier group. Receiver operating characteristic curve analysis of all seven myomiRs showed that the area under the curve (AUC) for miR-499, miR-133b, miR-1, miR-208b, and miR-133a exceeded 70.0%, and for miR-206 and miR-208a exceeded 60.0%. MiR-133b and miR-499 were significantly increased in all female carriers, even those with normal CK. AUC for the combination of all seven miRNAs was 87.2%. CK (OR 0.406, 95% CI 0.000–0.001, p < 0.0001) and miR-499 (OR 0.323, 95% CI 0.023–0.106, p = 0.003) were considered to be independent predictors for female carriers presence in the multivariable regression analysis model.Conclusions: MiR-133b and miR-499 are potentially useful biomarkers for female carriers with DMD/BMD (including those with normal CK). The combination of all seven serum miRNAs and their respective combinations with CK have better diagnostic value for female carriers than either CK or any separate miRNA.

背景：杜氏肌营养不良症/贝克肌营养不良症（DMD/BMD）是一种X连锁隐性致死性神经肌肉疾病。肌源微RNA（myomiRs）在横纹肌中的表达已被提议作为其潜在的生物标志物。血清肌酸激酶（CK）在临床实践中常被用作生物标志物，但其可靠性不足。本研究旨在评估血清肌源微RNA水平是否对检测具有正常或升高CK水平的女性DMD/BMD携带者具有诊断价值。方法：纳入34名女性携带者和33名年龄匹配的健康女性对照组。收集外周血样本，提取血清miRNA，通过定量实时聚合酶链反应测量miR-1、miR-133a、miR-133b、miR-206、miR-208a、miR-208b和miR-499。结果：与健康对照组相比，所有女性携带者的miR-1、miR-133a、miR-133b、miR-206、miR-208a、miR-208b和miR-499均上调。在女性携带者组中，miR-1与CK呈正相关（Spearman's rho = +0.406，p = 0.017）。所有七种肌源微RNA的受试者工作特征曲线分析显示，miR-499、miR-133b、miR-1、miR-208b和miR-133a的曲线下面积（AUC）均超过70.0%，而miR-206和miR-208a的AUC超过60.0%。即使在CK正常的情况下，所有女性携带者的miR-133b和miR-499也显著增加。所有七种miRNA的组合AUC为87.2%。在多变量回归分析模型中，CK（OR 0.406，95% CI 0.000–0.001，p < 0.0001）和miR-499（OR 0.323，95% CI 0.023–0.106，p = 0.003）被认为是女性携带者存在的独立预测因子。结论：miR-133b和miR-499可能是DMD/BMD女性携带者（包括CK正常的携带者）的有用生物标志物。所有七种血清miRNA及其与CK的组合，相较于CK或任何单一miRNA，均具有更高的诊断价值。