Headache and Neurological Pain Research Group, Vall d'Hebron Institute of Research (VHIR), Universitat Autònoma de Barcelona, Barcelona, Spain.
Neurology Department, Headache Unit, Vall d'Hebron University Hospital, Barcelona, Spain.
J Headache Pain. 2023 Feb 17;24(1):11. doi: 10.1186/s10194-023-01542-z.
Several studies have described potential microRNA (miRNA) biomarkers associated with migraine, but studies are scarcely reproducible primarily due to the heterogeneous variability of participants. Increasing evidence shows that disease-related intrinsic factors together with lifestyle (environmental factors), influence epigenetic mechanisms and in turn, diseases. Hence, the main objective of this exploratory study was to find differentially expressed miRNAs (DE miRNA) in peripheral blood mononuclear cells (PBMC) of patients with migraine compared to healthy controls in a well-controlled homogeneous cohort of non-menopausal women.
Patients diagnosed with migraine according to the International Classification of Headache Disorders (ICHD-3) and healthy controls without familial history of headache disorders were recruited. All participants completed a very thorough questionnaire and structured-interview in order to control for environmental factors. RNA was extracted from PBMC and a microarray system (GeneChip miRNA 4.1 Array chip, Affymetrix) was used to determine the miRNA profiles between study groups. Principal components analysis and hierarchical clustering analysis were performed to study samples distribution and random forest (RF) algorithms were computed for the classification task. To evaluate the stability of the results and the prediction error rate, a bootstrap (.632 + rule) was run through all the procedure. Finally, a functional enrichment analysis of selected targets was computed through protein-protein interaction networks.
After RF classification, three DE miRNA distinguished study groups in a very homogeneous female cohort, controlled by factors such as demographics (age and BMI), life-habits (physical activity, caffeine and alcohol consumptions), comorbidities and clinical features associated to the disease: miR-342-3p, miR-532-3p and miR-758-5p. Sixty-eight target genes were predicted which were linked mainly to enriched ion channels and signaling pathways, neurotransmitter and hormone homeostasis, infectious diseases and circadian entrainment.
A 3-miRNA (miR-342-3p, miR-532-3p and miR-758-5p) novel signature has been found differentially expressed between controls and patients with migraine. Enrichment analysis showed that these pathways are closely associated with known migraine pathophysiology, which could lead to the first reliable epigenetic biomarker set. Further studies should be performed to validate these findings in a larger and more heterogeneous sample.
已有多项研究描述了与偏头痛相关的潜在 microRNA（miRNA）生物标志物，但由于参与者的异质性差异较大，这些研究几乎无法重现。越来越多的证据表明，与疾病相关的内在因素以及生活方式（环境因素）会影响表观遗传机制，进而影响疾病。因此，这项探索性研究的主要目的是在绝经前女性这一经过严格控制的同质队列中，寻找偏头痛患者与健康对照者外周血单个核细胞（PBMC）中差异表达的 miRNA（DE miRNA）。
根据国际头痛疾病分类（ICHD-3）诊断为偏头痛的患者和无头痛疾病家族史的健康对照者被招募入组。所有参与者均完成了一份非常详细的问卷和结构化访谈，以控制环境因素。从 PBMC 中提取 RNA，并使用微阵列系统（GeneChip miRNA 4.1 Array 芯片，Affymetrix）来确定研究组之间的 miRNA 图谱。进行主成分分析和层次聚类分析以研究样本分布，并计算随机森林（RF）算法进行分类任务。为了评估结果的稳定性和预测错误率，通过所有程序运行了 bootstrap（.632+规则）。最后，通过蛋白质-蛋白质相互作用网络计算了选定靶标的功能富集分析。
通过 RF 分类，在一个经过严格控制的女性同质队列中，三个 DE miRNA 区分了研究组，这些因素包括人口统计学（年龄和 BMI）、生活习惯（体育活动、咖啡因和酒精摄入）、合并症和与疾病相关的临床特征：miR-342-3p、miR-532-3p 和 miR-758-5p。预测到 68 个靶基因，这些基因主要与丰富的离子通道和信号通路、神经递质和激素稳态、传染病和昼夜节律调节相关。
在对照组和偏头痛患者之间发现了一个由 3 个 miRNA（miR-342-3p、miR-532-3p 和 miR-758-5p）组成的新型差异表达 miRNA 特征。富集分析表明，这些途径与已知的偏头痛病理生理学密切相关，这可能导致第一个可靠的表观遗传生物标志物集。应进一步开展研究，在更大和更异质的样本中验证这些发现。
