Department of Experimental and Clinical Medicine, University of Florence, Firenze, Italy.
SOD Interdisciplinary Internal Medicine-Behçet Center and Lupus Clinic-Azienda Ospedaliero Universitaria Careggi, Azienda Ospedaliero Universitaria Careggi, Firenze, Italy.
Ann Rheum Dis. 2022 Mar;81(3):386-397. doi: 10.1136/annrheumdis-2021-220859. Epub 2021 Nov 29.
Behçet's syndrome (BS) is a rare systemic vasculitis often complicated by thrombotic events. Given the lack of validated biomarkers, BS diagnosis relies on clinical criteria.In search of novel biomarkers for BS diagnosis, we determined the profile of plasmatic circulating microRNAs (ci-miRNAs) in patients with BS compared with healthy controls (HCs).
ci-miRNA profile was evaluated by microarray in a screening cohort (16 patients with BS and 18 HCs) and then validated by poly(T) adaptor PCR (PTA-PCR) in a validation cohort (30 patients with BS and 30 HCs). Two disease control groups (30 patients with systemic lupus erythematosus (SLE) and 30 patients with giant cell arteritis (GCA) were also analysed.
From the microarray screening, 29 deregulated (differentially expressed (DE)) human ci-miRNAs emerged. A hierarchical cluster analysis indicated that DE ci-miRNAs clearly segregated patients from controls, independently of clinical features. PTA-PCR analysis on the validation cohort confirmed the deregulation of miR-224-5p, miR-206 and miR-653-5p. The combined receiver operating characteristic (ROC) curve analyses showed that such ci-miRNAs discriminate BS from HCs (and BS with active vs inactive disease), as well as BS from patients with SLE and GCA.The functional annotation analyses (FAAs) showed that the most enriched pathways affected by DE ci-miRNAs (ie, cell-matrix interaction, oxidative stress and blood coagulation) are related to thrombo-inflammatory mechanisms. Accordingly, the expression of the three ci-miRNAs from the validation cohort significantly correlated with leucocyte reactive oxygen species production and plasma lipid peroxidation.
The ci-miRNA profile identified in this study may represent a novel, poorly invasive BS biomarker, while suggesting an epigenetic control of BS-related thrombo-inflammation.
贝切特综合征（BS）是一种罕见的系统性血管炎，常伴有血栓形成事件。由于缺乏经过验证的生物标志物，BS 的诊断依赖于临床标准。为了寻找 BS 诊断的新型生物标志物，我们测定了 BS 患者与健康对照者（HCs）相比的血浆循环 microRNA（ci-miRNA）谱。
通过微阵列在筛查队列（16 例 BS 患者和 18 例 HCs）中评估 ci-miRNA 谱，然后在验证队列（30 例 BS 患者和 30 例 HCs）中通过 Poly（T）接头 PCR（PTA-PCR）进行验证。还分析了两个疾病对照组（30 例系统性红斑狼疮（SLE）患者和 30 例巨细胞动脉炎（GCA）患者）。
从微阵列筛选中，出现了 29 个下调（差异表达（DE））的人类 ci-miRNA。层次聚类分析表明，DE ci-miRNA 可清楚地将患者与对照者分开，与临床特征无关。验证队列的 PTA-PCR 分析证实了 miR-224-5p、miR-206 和 miR-653-5p 的下调。联合受试者工作特征（ROC）曲线分析表明，这些 ci-miRNA 可区分 BS 与 HCs（以及 BS 与活动期与非活动期疾病），以及 BS 与 SLE 和 GCA 患者。功能注释分析（FAAs）表明，DE ci-miRNA 最富集的途径（即细胞-基质相互作用、氧化应激和血液凝固）与血栓炎症机制有关。因此，验证队列中这三个 ci-miRNA 的表达与白细胞活性氧产生和血浆脂质过氧化显著相关。
本研究中鉴定的 ci-miRNA 谱可能代表一种新型、侵袭性低的 BS 生物标志物，同时提示 BS 相关血栓炎症存在表观遗传调控。
