MicroRNAs hsa-miR-99b, hsa-miR-330, hsa-miR-126 and hsa-miR-30c: Potential Diagnostic Biomarkers in Natural Killer (NK) Cells of Patients with Chronic Fatigue Syndrome (CFS)/ Myalgic Encephalomyelitis (ME).
MicroRNAs hsa-miR-99b, hsa-miR-330, hsa-miR-126 and hsa-miR-30c: Potential Diagnostic Biomarkers in Natural Killer (NK) Cells of Patients with Chronic Fatigue Syndrome (CFS)/ Myalgic Encephalomyelitis (ME).
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DOI：
10.1371/journal.pone.0150904
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Kerr JR
中科院分区：
文献类型：
--
作者：
Petty RD;McCarthy NE;Le Dieu R;Kerr JR
慢性疲劳综合征（CFS/ME）是一种病因不明的复杂多系统疾病，在全球人口的1%中引起衰弱症状。虽然大量的基因已被证明在CFS/ME患者中表现出改变的表达，但目前尚不清楚调节基因翻译的microRNA（miRNA）分子是否有助于疾病的发病机制。我们假设患者白细胞中microRNA表达的变化有助于CFS/ME病理学，因此可能代表可以在CFS/ME患者外周血中检测到的有用的诊断生物标志物。使用Ambion Bioarray V1分析CFS/ME患者和健康对照的外周血单核细胞（PBMC）中的miRNA表达。证明差异表达的miRNA通过qRT-PCR验证，然后在来自独立患者队列的分级血液白细胞亚群中复制。然后将通过这些实验鉴定的CFS/ME相关miRNA转染到原代NK细胞中，并进行基因表达分析以鉴定其基因靶标。微阵列分析鉴定了34种miRNA的差异表达，所有这些miRNA均上调。通过qRT-PCR证实了34种miRNA中的4种的表达变化。通过来自独立患者队列的细胞类型对PBMC样品进行分级，鉴定了NK细胞、B细胞和单核细胞中miRNA表达的变化，其中最显著的异常发生在NK细胞中。用hsa-miR-99 b或hsa-miR-330- 3 p转染原代NK细胞，导致与NK细胞活化一致的基因表达变化，但细胞毒性降低，表明缺陷的NK细胞功能有助于CFS/ME病理学。这项研究表明CFS/ME患者外周血单核细胞中微小RNA表达发生改变，这是潜在的诊断生物标志物。在NK细胞中鉴定出最大程度的miRNA失调，其靶点与细胞活化和改变的效应子功能一致。
Chronic Fatigue Syndrome (CFS/ME) is a complex multisystem disease of unknown aetiology which causes debilitating symptoms in up to 1% of the global population. Although a large cohort of genes have been shown to exhibit altered expression in CFS/ME patients, it is currently unknown whether microRNA (miRNA) molecules which regulate gene translation contribute to disease pathogenesis. We hypothesized that changes in microRNA expression in patient leukocytes contribute to CFS/ME pathology, and may therefore represent useful diagnostic biomarkers that can be detected in the peripheral blood of CFS/ME patients. miRNA expression in peripheral blood mononuclear cells (PBMC) from CFS/ME patients and healthy controls was analysed using the Ambion Bioarray V1. miRNA demonstrating differential expression were validated by qRT-PCR and then replicated in fractionated blood leukocyte subsets from an independent patient cohort. The CFS/ME associated miRNA identified by these experiments were then transfected into primary NK cells and gene expression analyses conducted to identify their gene targets. Microarray analysis identified differential expression of 34 miRNA, all of which were up-regulated. Four of the 34 miRNA had confirmed expression changes by qRT-PCR. Fractionating PBMC samples by cell type from an independent patient cohort identified changes in miRNA expression in NK-cells, B-cells and monocytes with the most significant abnormalities occurring in NK cells. Transfecting primary NK cells with hsa-miR-99b or hsa-miR-330-3p, resulted in gene expression changes consistent with NK cell activation but diminished cytotoxicity, suggesting that defective NK cell function contributes to CFS/ME pathology. This study demonstrates altered microRNA expression in the peripheral blood mononuclear cells of CFS/ME patients, which are potential diagnostic biomarkers. The greatest degree of miRNA deregulation was identified in NK cells with targets consistent with cellular activation and altered effector function.