Comprehensive analysis of microRNA profiles in multiple sclerosis including next-generation sequencing
Comprehensive analysis of microRNA profiles in multiple sclerosis including next-generation sequencing
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DOI：
10.1177/1352458513496343
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发表时间：
2014-03-01
影响因子：
5.8
通讯作者：
Ruprecht, Klemens
中科院分区：
文献类型：
--
作者：
Keller, Andreas;Leidinger, Petra;Ruprecht, Klemens
背景：微小RNA（miRNA）是具有基因调节功能的短的非编码RNA，其表达谱可作为疾病生物标志物。目的：本研究的目的是对临床孤立综合征（CIS）或复发缓解型多发性硬化症（RRMS）患者血液中的miRNA表达谱进行全面分析，包括下一代测序（NGS）。研究方法：通过NGS、微阵列分析和定量实时聚合酶链反应（qRT-PCR）分析来自CIS（n = 25）或RRMS（n = 25）的未经治疗患者和50名健康对照的全血样品中的miRNA表达。结果：在CIS/RRMS患者中，NGS和微阵列分析分别鉴定出38个和8个显著失调的miRNA。通过两种方法发现这些miRNAs中的三种显著上调（hsa-miR-16-2- 3 p）或下调（hsa-miR-20 a-5 p，hsa-miR-7-1- 3 p）。另外5个在NGS筛选中显著失调的miRNAs在微阵列分析中显示出相同的调控方向。qRT-PCR证实了所有八种miRNA的调控方向，并且对三种miRNA具有显著意义。结论：本研究鉴定了一组在CIS/RRMS中失调的miRNAs，并再次证实了先前报道的MS中hsa-miR-20 a-5 p的低表达。hsa-miR-20 a-5 p和其他经验证的miRNAs可能代表未来评估MS生物标志物的有希望的候选者，并且可能与该疾病的病理生理学相关。
Background: MicroRNAs (miRNAs) are short, noncoding RNAs with gene regulatory functions whose expression profiles may serve as disease biomarkers. Objective: The objective of this study was to perform a comprehensive analysis of miRNA expression profiles in blood of patients with a clinically isolated syndrome (CIS) or relapsing-remitting multiple sclerosis (RRMS) including next-generation sequencing (NGS). Methods: miRNA expression was analyzed in whole blood samples from treatment-naive patients with CIS (n = 25) or RRMS (n = 25) and 50 healthy controls by NGS, microarray analysis, and quantitative real-time polymerase chain reaction (qRT-PCR). Results: In patients with CIS/RRMS, NGS and microarray analysis identified 38 and eight significantly deregulated miRNAs, respectively. Three of these miRNAs were found to be significantly up- (hsa-miR-16-2-3p) or downregulated (hsa-miR-20a-5p, hsa-miR-7-1-3p) by both methods. Another five of the miRNAs significantly deregulated in the NGS screen showed the same direction of regulation in the microarray analysis. qRT-PCR confirmed the direction of regulation for all eight and was significant for three miRNAs. Conclusions: This study identifies a set of miRNAs deregulated in CIS/RRMS and reconfirms the previously reported underexpression of hsa-miR-20a-5p in MS. hsa-miR-20a-5p and the other validated miRNAs may represent promising candidates for future evaluation as biomarkers for MS and could be of relevance in the pathophysiology of this disease.