Sun Li, Tu Jianxin, Liu Cailong, Pan Axiao, Xia Xiaoru, Chen Xiaowei
Department of Rheumatology and Immunology, The First Affiliated Hospital of Wenzhou Medical University, Nanbaixiang, Ouhai District, Wenzhou, 325000, China.
Department of Orthopaedic Sports Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, China.
Inflammopharmacology. 2020 Apr;28(2):437-450. doi: 10.1007/s10787-019-00666-6. Epub 2019 Nov 28.
This study aimed to analyze long non-coding RNA (lncRNA) expression profiles in synovium tissue of patients with RA using RNA sequencing, and to further assess the clinical values of dysregulated lncRNAs in RA diagnosis and monitoring.
Thirty patients with RA who underwent knee arthroscopy and 30 controls with knee trauma who underwent surgery were consecutively enrolled and synovium tissue samples of both groups were obtained during surgery. In the exploration stage, lncRNA and mRNA expression profiles in three RA samples and three control samples were detected by RNA sequencing and bioinformatic analyses were then performed. In the validation stage, quantitative polymerase chain reaction (qPCR) was subsequently used to detect expression of five candidate lncRNAs in 30 patients with RA and 30 control patients.
A total of 349 lncRNAs and 1582 mRNAs were upregulated and 806 lncRNAs and 1295 mRNAs were downregulated in patients with RA compared with controls. Enrichment analyses revealed that these dysregulated lncRNAs and mRNAs were mainly involved in regulating immune response, leukocyte migration, complement activation, and B cell receptor signaling pathway. Subsequent qPCR validation discovered that lnc-AL928768.3 (P < 0.001) and lnc-AC091493.1 (P < 0.001) were elevated in patients with RA compared with controls and afford good predictive values for RA risk by receiver operating characteristic (ROC) curve analysis. Additionally, the two lncRNAs were positively associated with C-reactive protein level and disease activity score in 28 joints (ESR) (all P < 0.05).
Analysis of lncRNA expression profiles by sequencing reveals that lnc-AL928768.3 and lnc-AC091493.1 are novel biomarkers for RA risk and activity.
本研究旨在通过RNA测序分析类风湿关节炎（RA）患者滑膜组织中的长链非编码RNA（lncRNA）表达谱，并进一步评估失调的lncRNAs在RA诊断和监测中的临床价值。
连续纳入30例行膝关节镜检查的RA患者和30例因膝关节外伤接受手术的对照者，并在手术期间获取两组的滑膜组织样本。在探索阶段，通过RNA测序检测3例RA样本和3例对照样本中的lncRNA和mRNA表达谱，随后进行生物信息学分析。在验证阶段，随后使用定量聚合酶链反应（qPCR）检测30例RA患者和30例对照患者中5种候选lncRNAs的表达。
与对照组相比，RA患者中共有349种lncRNAs和1582种mRNAs上调，806种lncRNAs和1295种mRNAs下调。富集分析显示，这些失调的lncRNAs和mRNAs主要参与调节免疫反应、白细胞迁移、补体激活和B细胞受体信号通路。随后的qPCR验证发现，与对照组相比，RA患者中lnc-AL928768.3（P<0.001）和lnc-AC091493.1（P<0.001）升高，通过受试者工作特征（ROC）曲线分析对RA风险具有良好的预测价值。此外，这两种lncRNAs与C反应蛋白水平和28个关节疾病活动评分（ESR）呈正相关（均P<0.05）。
通过测序分析lncRNA表达谱发现，lnc-AL928768.3和lnc-AC091493.1是RA风险和活动的新型生物标志物。
