miR-326 和 miR-26a 是两种潜在的多发性硬化症缓解复发期诊断标志物。

miR-326 and miR-26a, two potential markers for diagnosis of relapse and remission phases in patient with relapsing-remitting multiple sclerosis.

机构信息

Division of Cellular and Molecular Biology, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran.

Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.

出版信息

Gene. 2014 Jul 10;544(2):128-33. doi: 10.1016/j.gene.2014.04.069. Epub 2014 Apr 30.

DOI:10.1016/j.gene.2014.04.069

PMID:24792898

Abstract

BACKGROUND

Multiple sclerosis is an inflammatory autoimmune disease widely characterized by myelin destruction of CNS. Th-17 cells, have been demonstrated to play a crucial role in pathogenesis of MS. MicroRNAs are a new class of non-coding RNAs that participate in post-transcriptional regulation of gene expression. Previous studies have reported a potential role of various miRNAs in induction of Th-17 differentiation and progress of autoimmune diseases. In recent years, it has been shown that miR-326 and miR-26a involved in progress of Th-17 and MS disease.

OBJECTIVE

To evaluate expression pattern of miR-326 and miR-26a in peripheral blood lymphocytes of relapsing-remitting MS patients during relapsing and remitting phases compared to healthy control subjects.

MATERIALS AND METHODS

Forty RR-MS patients of Isfahan population were diagnosed as relapsing (n=20) or remitting phase (n=20) patients according to clinical manifests and expression level of miR-26a and miR-326 was measured in these groups by quantitative real time PCR method compared to 20 healthy controls. In-silico molecular signaling pathway enrichment analysis was also performed on validated and predicted targets (targetome) of miR-26a by DAVID database to explore possible role of miR-26a in Th17 differentiation.

RESULTS

We observed up-regulation of both miR-326 and miR-26a in relapsing phase of multiple sclerosis patients compared with remitting phase (p value=0.0001) and healthy controls (p value=0.0091). ROC curve analysis confirmed valuable and precise potential of miR-326 to discriminate between relapsing and remitting phases of multiple sclerosis with specificity and sensitivity of 100% at a proposed optimum cutoff point. Furthermore, in-silico molecular signaling pathway enrichment analysis detected TGF-β signaling pathway as one of the most statistically relevant pathway with miR-26a targetome.

CONCLUSION

Our results confirmed potential of miR-326 as a diagnostic biomarker to discriminate between relapsing and remitting phases of multiple sclerosis disease. Similar expression pattern to miR-326 and in-silico molecular enrichment analysis altogether suggest an inducing role of miR-26a in differentiation of pathogenic Th17 cells during pathogenesis of multiple sclerosis by targeting major components of the TGF-β signaling pathway (i.e. SMAD4 and SMAD1) and disarrangement of this signaling pathway.

摘要

背景

多发性硬化症是一种广泛表现为中枢神经系统髓鞘破坏的炎症性自身免疫性疾病。Th-17 细胞已被证明在多发性硬化症的发病机制中起关键作用。 microRNAs 是一类新的非编码 RNA，参与基因表达的转录后调控。先前的研究报道了各种 microRNAs 在诱导 Th-17 分化和自身免疫性疾病进展中的潜在作用。近年来，研究表明 miR-326 和 miR-26a 参与 Th-17 和 MS 疾病的进展。

目的

评估复发缓解型多发性硬化症患者在复发和缓解期外周血淋巴细胞中 miR-326 和 miR-26a 的表达模式与健康对照组相比。

材料和方法

根据临床表现，将 40 名来自伊斯法罕的 RR-MS 患者诊断为复发期（n=20）或缓解期（n=20）患者，采用定量实时 PCR 法测量这些组中 miR-26a 和 miR-326 的表达水平，并与 20 名健康对照者进行比较。还通过 DAVID 数据库对 miR-26a 的验证和预测靶标（靶标组）进行了基于分子的信号通路富集分析，以探讨 miR-26a 在 Th17 分化中的可能作用。

结果

与缓解期（p 值=0.0001）和健康对照组（p 值=0.0091）相比，我们观察到多发性硬化症患者在复发期时 miR-326 和 miR-26a 均上调。ROC 曲线分析证实，miR-326 具有区分多发性硬化症复发和缓解期的高价值和精确潜力，在建议的最佳截断点处，特异性和敏感性均为 100%。此外，基于分子的信号通路富集分析检测到 TGF-β 信号通路是 miR-26a 靶标组中最具统计学意义的通路之一。

结论

我们的研究结果证实，miR-326 有潜力作为诊断生物标志物，以区分多发性硬化症疾病的复发和缓解期。与 miR-326 相似的表达模式和基于分子的富集分析表明，miR-26a 通过靶向 TGF-β 信号通路的主要成分（即 SMAD4 和 SMAD1）和干扰该信号通路，在多发性硬化症发病机制中诱导致病性 Th17 细胞分化中起诱导作用。