Li Guoqing, Chen Hongyi, Shen Jiacheng, Ding Yimin, Chen Jingqiong, Zhang Yongbin, Tang Mingrui, Xu Nan, Fang Yuxuan
Department of Rheumatology and Immunology, Affiliated Hospital of Yangzhou University, Yangzhou University, No. 368 Hanjiang Middle Road, Yangzhou, Jiangsu, 225000, China.
J Orthop Surg Res. 2025 Jan 29;20(1):109. doi: 10.1186/s13018-025-05494-9.
Rheumatoid arthritis (RA), a chronic inflammatory joint disease causing permanent disability, involves exosomes, nanosized mammalian extracellular particles. Circular RNA (circRNA) serves as a biomarker in RA blood samples. This research screened differentially expressed circRNAs in RA patient plasma exosomes for novel diagnostic biomarkers. In this study, samples of RA patients with insufficient response to methotrexate (MTX-IR), combined use of tumor necrosis factor inhibitors (TNFi) were followed up for half a year, and 56 circRNA samples of self-test data were stratified into training, testing, and external validation cohorts according to whether American College of Rheumatology 20% improvement criteria (ACR20) was achieved. A diagnostic xgboost model was developed using common hub genes identified by random forest and least absolute shrinkage and selection operator (LASSO), with intersection genes derived from overlapping machine learning-selected genes. Diagnostic performance evaluated via receiver operating characteristic (ROC) curves using pROC for area under the curve (AUC). Optimal LASSO model with 4 circRNAs determined, with AUC > 0.6 for key genes. The model validation performed well on the test set, but not significantly on the validation set. Then, circRNA screening was performed in combination with clinical data, and cross-validation identified hsa-circ0002715, hsa-circ0001946, hsa-circ0000836, and rheumatoid factor (RF) as key genes, among which hsa-circ0002715 and hsa-circ0001946 were emphasized as key markers on the training set. In addition, the morphology and size of exosomes and the expression of CD9 and CD81 verified the successful extraction of exosomes. The qPCR analysis of plasma exosomes in TNFi patients found that the expression of hsa-circ0002715 was higher than that in patients who didn't reach ACR20, and the expression of hsa-circ0001946 was lower than that in patients who didn't reach ACR20. The above studies suggested that hsa-circ0002715 and hsa-circ0001946 may become markers for predicting MTX-IR RA patients and TNFi precision treatment.
类风湿性关节炎（RA）是一种导致永久性残疾的慢性炎症性关节疾病，涉及外泌体，即纳米级的哺乳动物细胞外颗粒。环状RNA（circRNA）可作为RA血液样本中的生物标志物。本研究筛选了RA患者血浆外泌体中差异表达的circRNA，以寻找新的诊断生物标志物。在本研究中，对甲氨蝶呤反应不足（MTX-IR）的RA患者样本联合使用肿瘤坏死因子抑制剂（TNFi）进行了半年的随访，并根据是否达到美国风湿病学会20%改善标准（ACR20）将56个circRNA自检数据样本分层为训练、测试和外部验证队列。使用通过随机森林和最小绝对收缩和选择算子（LASSO）确定的常见枢纽基因开发了一种诊断性XGBoost模型，其交叉基因源自重叠的机器学习选择基因。使用pROC通过受试者操作特征（ROC）曲线评估诊断性能以计算曲线下面积（AUC）。确定了具有4个circRNA的最佳LASSO模型，关键基因的AUC>0.6。该模型在测试集上表现良好，但在验证集上表现不显著。然后，结合临床数据进行circRNA筛选，交叉验证确定hsa-circ0002715、hsa-circ0001946、hsa-circ0000836和类风湿因子（RF）为关键基因，其中hsa-circ0002715和hsa-circ0001946在训练集上被强调为关键标志物。此外，外泌体的形态和大小以及CD9和CD81的表达验证了外泌体的成功提取。对TNFi患者血浆外泌体的qPCR分析发现，hsa-circ000271�的表达高于未达到ACR20的患者，而hsa-circ0001946的表达低于未达到ACR20的患者。上述研究表明，hsa-circ0002715和hsa-circ0001946可能成为预测MTX-IR RA患者和TNFi精准治疗的标志物。
