Wang Xing-Zhi, Li Shuo, Liu Yun, Cui Gui-Yun, Yan Fu-Ling
School of Medicine, Southeast University, Nanjing, Jiangsu, 210009, People's Republic of China.
Department of Neurology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu, 221006, People's Republic of China.
J Inflamm Res. 2022 Jul 17;15:4087-4104. doi: 10.2147/JIR.S368417. eCollection 2022.
Accumulating evidence suggests that circular RNAs (circRNAs) are involved in immune and inflammatory processes after acute ischemic stroke (AIS). However, the roles of circRNA-mediated competing endogenous RNA (ceRNA) in modulating immune inflammation of AIS have not yet been determined. This study aimed to construct a circRNA-mediated immune-related ceRNA network and identify novel circRNAs in AIS.
Microarray data were downloaded from the GEO database and further analysed by R software. Then, we constructed a circRNA-mediated ceRNA network based on interaction information from the bioinformatics database. A topological property analysis of the ceRNA network was conducted to screen novel circRNAs. Finally, we further applied quantitative real-time polymerase chain reaction (qRT-PCR) to two independent sets.
We constructed an AIS immune-related ceRNA (AISIRC) network containing immune-related genes (IRGs), miRNAs, and circRNAs. Additionally, we extracted the subnetwork from the AISIRC network and screened six immune-related circRNAs. After identification and validation, we finally confirmed that plasma levels of circPTP4A2 and circTLK2 were significantly increased in AIS patients compared with both healthy control subjects (HCs) and transient ischemic attack (TIA) patients. Logistic regression and receiver-operating characteristic (ROC) curve analyses demonstrated that these two circRNAs may function as predictive and discriminative biomarkers for AIS. We also confirmed that plasma levels of circPTP4A2 were elevated in TIA patients compared with HCs and might be an independent risk factor for predicting TIA. Longitudinal analysis of circRNA expression up to 90 days after AIS indicated that the ability of circPTP4A2 and circTLK2 to monitor AIS dynamics was highly desirable.
In summary, the circRNA-mediated immune-related ceRNA network was successfully constructed, and two circulating circRNAs (circPTP4A2 and circTLK2) improved sensitivity for the diagnosis of AIS and could be considered diagnostic biomarkers.
越来越多的证据表明，环状RNA（circRNA）参与急性缺血性卒中（AIS）后的免疫和炎症过程。然而，circRNA介导的竞争性内源性RNA（ceRNA）在调节AIS免疫炎症中的作用尚未确定。本研究旨在构建circRNA介导的免疫相关ceRNA网络，并鉴定AIS中的新型circRNA。
从基因表达综合数据库（GEO数据库）下载微阵列数据，并使用R软件进行进一步分析。然后，我们根据生物信息学数据库中的相互作用信息构建了一个circRNA介导的ceRNA网络。对ceRNA网络进行拓扑性质分析以筛选新型circRNA。最后，我们在两个独立的数据集中进一步应用了定量实时聚合酶链反应（qRT-PCR）。
我们构建了一个包含免疫相关基因（IRG）、微小RNA（miRNA）和circRNA的AIS免疫相关ceRNA（AISIRC）网络。此外，我们从AISIRC网络中提取了子网，并筛选出六个免疫相关的circRNA。经过鉴定和验证，我们最终证实，与健康对照受试者（HC）和短暂性脑缺血发作（TIA）患者相比，AIS患者血浆中circPTP4A2和circTLK2的水平显著升高。逻辑回归和受试者工作特征（ROC）曲线分析表明，这两种circRNA可能作为AIS的预测和鉴别生物标志物。我们还证实，与HC相比，TIA患者血浆中circPTP4A2水平升高，可能是预测TIA的独立危险因素。对AIS后长达90天的circRNA表达进行纵向分析表明，circPTP4A2和circTLK2监测AIS动态的能力非常理想。
总之，成功构建了circRNA介导的免疫相关ceRNA网络，两种循环circRNA（circPTP4A2和circTLK2）提高了AIS诊断的敏感性，可被视为诊断生物标志物。
