He Hengqian, Zhang Juntao, Gong Weikun, Liu Mengyun, Liu Hao, Li Xiaoyong, Wu Yufei, Lu Qinkang
Department of Ophthalmology, The Affiliated People's Hospital of Ningbo University, Ningbo, China.
Center for Disease Control and Prevention of Yinzhou District, Ningbo, China.
Front Genet. 2022 Feb 17;13:833573. doi: 10.3389/fgene.2022.833573. eCollection 2022.
Circular RNAs (circRNAs), a class of non-coding and undegradable RNAs, play many pathological functions by acting as miRNA sponges, interacting with RNA-binding proteins, and others. The recent literature indicates that circRNAs possess the advanced superiority for the early screening of diabetic retinopathy (DR). CircRNA sources of peripheral blood mononuclear cells (PBMCs) from healthy controls ( = 4), diabetes mellitus patients (DM) ( = 4), and DR patients ( = 4) were extracted for circular RNA microarray analysis. Enriched biological modules and signaling pathways were analyzed by Gene Ontology Enrichment and Kyoto Encyclopedia of Genes and Genomes analysis, respectively. Real-time quantitative reverse transcription PCR (RT-qPCR) was performed to validate differentiated levels of several circRNAs (fold change ≥2, < .05) in different groups of healthy control subjects ( = 20), DM patients ( = 60), and DR patients ( = 42). Based on our clinical data from DR, the diagnostic performance of candidate circRNAs was measured by operating characteristic curves (ROCs). Subsequently, their circRNA-miRNA networks were constructed by bioinformatics analysis. : Circular RNA microarray analysis was performed, and 2,452 and 289 circRNAs were screened with differential expression in DR patients compared to healthy controls and DM patients, respectively. Enrichment analyses showed that circRNAs in DR patients were enriched in extracellular matrix (ECM)-receptor interaction and focal adhesion pathways. The top 5 differential circRNAs in circRNA microarray analysis were subsequently quantified and verified by RT-qPCR. Consistently, a significant 2.2-fold reduction of hsa_circ_0095008 and 1.7-fold increase in hsa_circ_0001883 were identified in DR patients compared to DM patients. Meanwhile, the area under curves of hsa_circ_0095008 and hsa_circ_0001883 were 0.6710 (95% CI, 0.5646-0.7775) ( = 0.003399) and 0.6071 (95% CI, 0.4953-0.7189) ( = 0.06644), respectively, indicating a good diagnostic value. Our study provided a new sight for the pathological mechanism of DR and revealed the potential value of hsa_circ_0095008 and hsa_circ_0001883 as diagnostic biomarkers for the early diagnosis of DR patients.
环状RNA(circRNAs)是一类非编码且不可降解的RNA,通过充当微小RNA(miRNA)海绵、与RNA结合蛋白相互作用等发挥多种病理功能。最近的文献表明,circRNAs在糖尿病视网膜病变(DR)的早期筛查中具有显著优势。提取了来自健康对照者(n = 4)、糖尿病(DM)患者(n = 4)和DR患者(n = 4)的外周血单个核细胞(PBMCs)的circRNA来源,用于环状RNA微阵列分析。分别通过基因本体论富集分析和京都基因与基因组百科全书分析,对富集的生物模块和信号通路进行分析。采用实时定量逆转录PCR(RT-qPCR)验证了不同组健康对照者(n = 20)、DM患者(n = 60)和DR患者(n = 42)中几种circRNAs(倍数变化≥2,P < 0.05)的差异水平。基于我们的DR临床数据,通过操作特征曲线(ROC)测量候选circRNAs的诊断性能。随后,通过生物信息学分析构建它们的circRNA-miRNA网络。结果:进行了环状RNA微阵列分析,与健康对照者和DM患者相比,分别筛选出2452个和289个在DR患者中差异表达的circRNAs。富集分析表明,DR患者中的circRNAs富集于细胞外基质(ECM)-受体相互作用和粘着斑通路。随后对环状RNA微阵列分析中前5个差异circRNAs进行定量,并通过RT-qPCR验证。与DM患者相比,DR患者中hsa_circ_0095008显著降低2.2倍,hsa_circ_0001883增加1.7倍。同时,hsa_circ_0095008和hsa_circ_0001883的曲线下面积分别为0.6710(95%CI,0.5646 - 0.7775)(P = 0.003399)和0.6071(95%CI,0.4953 - 0.7189)(P = 0.06644),表明具有良好的诊断价值。我们的研究为DR的病理机制提供了新的见解,并揭示了hsa_circ_0095008和hsa_circ_0001883作为DR患者早期诊断生物标志物的潜在价值。