Analysis of serum circulating MicroRNAs level in Malaysian patients with gestational diabetes mellitus

Abstract

Gestational diabetes mellitus (GDM) is a severe global issue that requires immediate attention. MicroRNA expression abnormalities are possibly disease-specific and may contribute to GDM pathological processes. To date, there is limited data on miRNA profiling in GDM, especially that involves a longitudinal study. Here, we performed miRNA expression profiling in the entire duration of pregnancy (during pregnancy until parturition and postpartum) using a miRNA- polymerase chain reaction array (miRNA-PCRArray) and in-silico analysis to identify unique miRNAs expression and their anticipated target genes in Malay maternal serum. MiRNA expression levels and their unique potential as biomarkers were explored in this work. In GDM patients, the expression levels of hsa-miR-193a, hsa-miR-21, hsa-miR-23a, and hsa-miR-361 were significantly increased, but miR-130a was significantly downregulated. The area under the curve (AUC) and receiver operating characteristic (ROC) curve study demonstrated that hsa-miR-193a (AUC = 0.89060 ± 04,470, P = 0.0001), hsa-miR-21 (AUC = 0.89500 ± 04,411, P = 0.0001), and miR-130a (AUC = 0.6939 ± 0.05845, P = 0.0025) had potential biomarker features in GDM. In-silico analysis also revealed that KLF (Kruppel-Like family of transcription factor), ZNF25 (Zinc finger protein 25), AFF4 (ALF transcription elongation factor 4), C1orf143 (long intergenic non-protein coding RNA 2869), SRSF2 (serine and arginine rich splicing factor 2), and ZNF655 (Zinc finger protein 655) were prominent genes targeted by the common nodes of miR23a, miR130, miR193a, miR21, and miR361.Our findings suggest that circulating microRNAs in the first trimester has the potential for GDM screening in the Malay population.

Figure 1

Scatter plot analysis of the relative expression of thirty-seven miRNAs compared between control and GDM patients. (A) = first trimester, (B) = second trimester (C) = third trimester (D) = post-partum. Color scheme: red color indicates upregulated miRNAs, while blue indicates downregulated miRNAs. The miRNAs that are significantly altered between the two groups (control and GDM) are located above. The diagonal black line corresponds to P < 0.05. Analysis was performed using online statistical analysis by Qiagen.

Figure 2

Dysregulated expression pattern of microRNAs in First (A), Second (B), Third (C) and postpartum (D) in GDM patients. Charts were derived from Graf Pad Prism version8. P-value calculated using non-parametric analytical test.

Figure 3

ROC curve analysis. The figure shows the comparison of five dysregulated microRNAs and their specific AUC based on their ΔCt distribution in the first trimester of pregnancy (A–E). Cumulative Graff of ROC analysis of three combined microRNAs and Binary logistic regression (BLR) analysis (E and F). The ROC curve is derived from Graf Pad Prism version 8. Normality distribution of samples was performed using Shapiro–Wilk test. P-value calculated using non-parametric analytical test.

Figure 4

In-silico analysis of dysregulated microRNAs in the first trimester. Green and purple colors indicate dysregulated microRNAs and their common target genes, respectively. ZNF25, RANBP17, CLORF43, ZNF655, AFF4, KLF and SRSF2 were targeted in a common node of microRNAs. Interaction pathway provided by Cytoscape tool version 3.7.1.