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首页> 外文期刊>BMC Medical Genomics >Erythrocyte microRNA sequencing reveals differential expression in relapsing-remitting multiple sclerosis
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Erythrocyte microRNA sequencing reveals differential expression in relapsing-remitting multiple sclerosis

机译：红细胞microRNA测序揭示复发性多发性硬化症中的差异表达

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There is a paucity of knowledge concerning erythrocytes in the aetiology of Multiple Sclerosis (MS) despite their potential to contribute to disease through impaired antioxidant capacity and altered haemorheological features. Several studies have identified an abundance of erythrocyte miRNAs and variable profiles associated with disease states, such as sickle cell disease and malaria. The aim of this study was to compare the erythrocyte miRNA profile of relapsing-remitting MS (RRMS) patients to healthy sex- and age-matched controls. Erythrocytes were purified by density-gradient centrifugation and RNA was extracted. Following library preparation, samples were run on a HiSeq4000 Illumina instrument (paired-end 100?bp sequencing). Sequenced erythrocyte miRNA profiles (9 patients and 9 controls) were analysed by DESeq2. Differentially expressed miRNAs were validated by RT-qPCR using miR-152-3p as an endogenous control and replicated in a larger cohort (20 patients and 18 controls). After logarithmic transformation, differential expression was determined by two-tailed unpaired t-tests. Logistic regression analysis was carried out and receiver operating characteristic (ROC) curves were generated to determine biomarker potential. A total of 236 erythrocyte miRNAs were identified. Of twelve differentially expressed miRNAs in RRMS two showed increased expression (adj. p?
机译：尽管多发性硬化症（MS）的病因可能会通过抗氧化能力受损和血液流变学特征改变而促成疾病，但对红细胞的了解却很少。几项研究已经确定了丰富的红细胞miRNA和与疾病状态（例如镰状细胞病和疟疾）相关的可变图谱。这项研究的目的是比较复发缓解型MS（RRMS）患者与健康的性别和年龄相匹配的对照组的红细胞miRNA谱。通过密度梯度离心纯化红细胞并提取RNA。文库制备后，将样品在HiSeq4000 Illumina仪器上运行（配对末端100 bp测序）。通过DESeq2分析测序的红细胞miRNA图谱（9例患者和9例对照）。使用miR-152-3p作为内源对照，通过RT-qPCR验证了差异表达的miRNA，并在更大的队列中复制（20名患者和18名对照）。对数转换后，通过两尾未配对t检验确定差异表达。进行逻辑回归分析，并生成接收器操作特征（ROC）曲线以确定生物标志物的潜力。总共鉴定出236种红细胞miRNA。在RRMS中的十二个差异表达的miRNA中，有两个显示出增加的表达（调节p <0.05）。跨差异表达的miRNA仅有明显的倍数变化。与健康对照相比，RT-qPCR证实了RRMS中miR-30b-5p（0.61倍，p 0.05）和miR-3200-3p（0.36倍，p 0.01）的差异表达。 miR-3200-5p的相对表达（0.66倍，NS p？=？0.096）也具有重要意义。 MiR-3200-5p与通过录音的认知屏幕测得的认知呈正相关（r = 0.60; p <0.01）。作为单个miRNA，MiR-3200-3p表现出最大的生物标志物潜力（准确度≥75.5％，p≤0.01，灵敏度≤72.7％，特异性≤84.0％）。将miR-3200-3p，miR-3200-5p和miR-30b-5p组合到复合生物标记物中，可将准确性提高到83.0％（p <0.05），灵敏度提高到77.3％，特异性提高到88.0％。这是第一个报告RRMS中红细胞miRNA差异的研究。尽管尚需阐明miRNA在红细胞中的作用，但红细胞miRNA的差异表达可能会被用作生物标记物，并应进一步研究其对MS病理学和认知的潜在影响。

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