Potential value of lncRNAs as a biomarker for proliferative diabetic retinopathy.

To investigate the differences in lncRNAs expression in whole blood between diabetic retinopathy (DR) patients and healthy subjects, and to evaluate the potential value of lncRNAs as a diagnostic biomarker for proliferative diabetic retinopathy (PDR). A series of 34 PDR patients, 34 patients with non-proliferative DR (NPDR) and 34 healthy participants were enroled. Differentially expressed lncRNAs were demonstrated using high-throughput sequencing and validated using qRT-PCR. Gene Ontology (GO) was performed to explore the possible biological function of the differentially expressed lncRNAs. lncRNA/mRNA coexpression network was built to determine the targets of differentially expressed lncRNAs. Receiver operating characteristic (ROC) analysis was utilized to evaluate the diagnostic value of lncRNAs for PDR. We identified 175 and 179 differentially expressed lncRNAs in PDR patients compared with control samples and NPDR patients, respectively. GO analysis showed that the various metabolic processes were possibly influenced by these dysregulated lncRNAs. Using the differently expressed lncRNAs data, we further identified 82 overlapping lncRNAs in PDR patients with NPDR and control subjects. Part of these overlapping lncRNAs was significantly correlated with nuclear factor kappa B (NF-κB) and Wnt signal pathways. ROC curves were constructed for two upregulated lncRNAs and the ROC analysis indicated that both of them had potential diagnostic value and could distinguish PDR from control subjects and NPDR patients. LncRNAs expression was altered in PDR patients compared with NPDR and control subjects. Moreover, it provides a resource that lncRNAs might be novel diagnostic and prognostic biomarker for PDR.

Liu B ，Cong C ，Ma Y ，Ma X ，Zhang H ，Wang J ... -  《-》

Diagnostic and prognostic role of serum miR-20b, miR-17-3p, HOTAIR, and MALAT1 in diabetic retinopathy.

Noncoding RNAs are emerging biomarkers for many diseases including diabetic retinopathy (DR). This study aimed to measure the expression levels of serum miR-20b, miR-17-3p, HOTAIR, and MALAT1 in DR patients. A total of 80 patients diagnosed as type 2 diabetes (T2D) and 81 healthy subjects were recruited in this study. T2D patients were divided into three groups: nondiabetic retinopathy (NDR) group (30 patients), nonproliferative diabetic retinopathy (NPDR) group (30 patients), and proliferative diabetic retinopathy (PDR) group (20 patients). Quantitative real-time polymerase chain reaction (PCR) was used to assess the expression of serum miR-20b, miR-17-3p, HOTAIR, and MALAT1. We found a significant decrease in serum miR-20b and a significant increase in serum HOTAIR and MALAT1 in NDR patients compared to healthy subjects. Also, we revealed a significant decrease in serum miR-20b and miR-17-3p and a significant increase in serum HOTAIR and MALAT1 in each of NPDR and PDR groups when compared with healthy subjects. Furthermore, we reported a significant decrease in miR-20b and miR-17-3p and a significant increase in HOTAIR and MALAT1in DR as well as in PDR patients when compared with NDR patients. However, on comparing NPDR with NDR patients, no significant difference was observed regarding the expression levels of miR-20b and miR-17-3p, in contrast, significant elevation of serum HOTAIR and MALAT1 was found in NPDR. Moreover, we observed a significant decrease in serum miR-20b and miR-17-3p and a significant increase in serum HOTAIR and MALAT1 in PDR group relative to NPDR group. Receiver operating characteristic (ROC) curve was used for evaluating the diagnostic value of the examined serum noncoding RNAs as novel biochemical indicators detecting severity of DR. Our analyses suggested that the examined serum noncoding RNAs may discriminate DR (PDR and NPDR) from NDR. Furthermore, these noncoding RNAs (less importantly miR-17) can be used as promising novel biomarkers for prediction DR severity, distinguishing PDR from NPDR patients. We can conclude that serum miR-20b, miR-17-3p, HOTAIR, and MALAT1 may be used as noninvasive biomarkers for screening of DR and early diagnosis of PDR. © 2018 IUBMB Life, 71(3):310-320, 2019.

Shaker OG ，Abdelaleem OO ，Mahmoud RH ，Abdelghaffar NK ，Ahmed TI ，Said OM ，Zaki OM ... -  《-》

Effect of intravitreal conbercept treatment on the expression of Long Noncoding RNAs and mRNAs in Proliferative Diabetic Retinopathy Patients.

To evaluate the effect of conbercept on the expression of long noncoding RNAs (lncRNAs) and mRNAs in the fibrovascular membranes of proliferative diabetic retinopathy (PDR) patients. Twenty patients, diagnosed with PDR, who underwent pars plana vitrectomy (PPV), were recruited for this study. Ten patients were treated for PPV alone (Control Group), and the others received conbercept injections before PPV (Treated Group). The fibrovascular membranes were harvested during surgery. Expression of lncRNAs and mRNAs in the membranes was tested using lncRNA Arrays. Bioinformatics analyses were performed to identify the related biological modules and pathways of the differentially expressed genes. A lncRNA/mRNA coexpression network was built to identify the correlations between lncRNAs and mRNAs. Real-time PCR was conducted to verify the microarray results. We identified 427 differentially expressed lncRNAs, of which 263 were upregulated and 164 were downregulated. Gene ontology (GO) analysis indicated that these lncRNAs-coexpressed mRNAs targeted various metabolic processes, especially the gluconeogenesis. Kyoto Encyclopaedia of Genes and Genomes (KEGG) results indicated that 16 pathways had significant differences in gene expression, including gluconeogenesis, HIF-1 signalling pathway, NOD-like receptor pathway, etc. The lncRNA/mRNA coexpression network revealed that many differentially expressed lncRNAs were enriched in the HIF-1, TNF-α and NOD-like receptor pathways. LincRNAs were the largest category and further bioinformatics analysis implied that these lincRNAs-coexpressed mRNAs were mainly involved in PDR-related biological processes and pathological pathways. Conbercept treatment can change the expression profiles of lncRNAs and mRNAs in the fibrovascular membranes of PDR patients. A complete understanding of the relationship between lncRNAs and anti-VEGF drugs may contribute to new therapeutic regimen for PDR.

Wang J ，Gao X ，Liu J ，Wang J ，Zhang Y ，Zhang T ，Zhang H ... -  《-》

Discovery and validation of hsa_circ_0001953 as a potential biomarker for proliferative diabetic retinopathy in human blood.

This study aimed to determine whether circular RNAs (circRNAs) in whole blood could be served as novel non-invasive biomarkers for proliferative diabetic retinopathy (PDR). This retrospective cross-sectional study comprised 34 healthy participants, 34 PDR patients and 34 non-proliferative DR (NPDR) patients. High-throughput whole transcriptome sequencing was performed to explore the expression profile of circRNAs in the whole blood, and the candidate circRNAs were validated by quantitative real-time polymerase chain reaction (qRT-PCR). Receiver operating characteristic (ROC) analysis evaluated the ability of these candidate circRNAs in discriminating PDR patients from NPDR patients and healthy subjects. Finally, the networks of circRNA-miRNA-mRNA based on the candidate circRNAs were constructed. Using sequencing and qRT-PCR, hsa_circ_0001953 was found to be elevated in PDR patients in contrast with the other two groups. Statistical analysis showed that the expression levels of hsa_circ_0001953 in PDR patients were positively related to the duration of diabetes and HbAc1. Receiver operating characteristic (ROC) curve analysis revealed that hsa_circ_0001953 was associated with a high diagnostic accuracy in discriminating PDR patients from NPDR patients and healthy controls, resulting in an area under the curve (AUC) of 0.87 and 0.92, respectively. The circRNA-miRNA-target gene networks for hsa_circ_0001953 showed that hsa_circ_0001953 could interact with dozens of miRNAs and some targeted mRNAs have been potentially involved in the pathogenesis of diabetes. The present findings indicate that hsa_circ_0001953 in the whole blood may serve as a novel diagnostic biomarker and potential therapeutic target for PDR.

Wu Z ，Liu B ，Ma Y ，Chen H ，Wu J ，Wang J ... -  《-》

High level of lncRNA NR2F1-AS1 predict the onset and progression of diabetic retinopathy in type 2 diabetes.

This study aimed to investigate whether dysregulation of NR2F1-AS1 is associated with the T2D onset and DR progression. A total of 269 individuals diagnosed with T2D were recruited in this study, including 111 individuals with uncomplicated T2D and 158 individuals with nonproliferative diabetic retinopathy (NPDR). A total of 48 among the NPDR individuals developed into proliferative diabetic retinopathy (PDR), excepting that 6 individuals were dropped out. RT-qPCR was used to determine the expression of NR2F1-AS1 level in blood. The correlations between NR2F1-AS1 expression and other parameters were tested by Pearson correlation. Receiver operating characteristic (ROC) curves were plotted for evaluating the diagnostic value of NR2F1-AS1 as a biochemical indicator detecting T2D and PDR. Kaplan-Meier method and Cox multivariable analysis was used to evaluate the predictive power of NR2F1-AS1 for incidence of PDR. A significant increase in blood NR2F1-AS1 in uncomplicated T2D, NPDR, and PDR patients was found when compared to healthy subjects. Significant correlations between NR2F1-AS1 expression and the level of fasting glucose, 2-h postprandial blood glucose, and HbA1c were revealed. Furthermore, NR2F1-AS1 represented distinguishing ability in T2D individuals from healthy subjects and PDR individuals from NPDR individuals. NR2F1-AS1 also showed strong predictive ability for PDR. The examination of blood NR2F1-AS1 may be used as a noninvasive biomarker for screening of T2D and early diagnosis of DR. NR2F1-AS1 may also be used as a promising novel biomarker for prediction PDR. NR2F1-AS1 may play a role in the progression of DR by moderating EndMT.

Ji X ，Sun J ，Wang Z 《-》