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This study identifies and validates circulating microRNAs as biomarkers for early detection of gestational diabetes mellitus (GDM).

February 8, 2026Journal of Endocrinological Investigation0 citationsOpen Access

Circulating miR-326 and miR-532-3p: novel biomarkers for early gestational diabetes mellitus detection with high diagnostic accuracy

HLHui-Zhen LinFujian Medical University ZLZhikun LiangGuangzhou Mechanical Engineering Research Institute (China)JHJun-Ju HuangGuangzhou Mechanical Engineering Research Institute (China)

Key Points

This study identifies and validates circulating microRNAs as biomarkers for early detection of gestational diabetes mellitus (GDM).
Conducted a two-phase cross-sectional study with 55 GDM patients and 55 healthy controls.
Used RNA sequencing to discover differentially expressed miRNAs in blood samples.
Validated findings with reverse transcription and quantitative PCR for the entire cohort.
Performed bioinformatics analyses to explore functional roles of identified miRNAs.
Significant upregulation of miR-326 and miR-532-3p in GDM patients confirmed by qPCR.
Found strong negative correlations between miRNA levels and glucose levels post-meal.
Achieved excellent diagnostic performance with AUCs of 0.95 for miR-326 and 0.96 for miR-532-3p.
Functional analyses linked these miRNAs to critical metabolic signaling pathways.

Abstract

Abstract Purpose Gestational diabetes mellitus (GDM) significantly threatens maternal and fetal health, necessitating early and accurate diagnostic tools. This study aimed to identify and validate circulating microRNAs (miRNAs) as novel biomarkers for GDM. Methods A two-phase cross-sectional study enrolled 55 GDM patients and 55 matched healthy pregnant controls. In the discovery phase, RNA sequencing (RNA-seq) of peripheral blood RNA from a randomly selected subset (5 GDM patients, 5 controls) identified differentially expressed miRNAs. The validation phase employed reverse transcription and quantitative PCR (qPCR) in the entire cohort to confirm identified miR-326 and miR-532-3p expression. Bioinformatics analyses (Gene Ontology GO and Kyoto Encyclopedia of Genes and Genomes KEGG) investigated their functional roles using a consensus-based approach across multiple databases. Results RNA-seq revealed significant upregulation of miR-326 (mean normalized counts = 14.59, P < 0.05; log 2 Fold Change = 0.93, P < 0.01) and miR-532-3p (mean normalized counts = 46.45, P < 0.05; log 2 Fold Change = 0.66, P < 0.01) in GDM patients, a finding corroborated by cross-referencing with the Gene Expression Omnibus (GEO) database. qPCR validation confirmed significantly higher expression for both miRNAs in GDM ( P < 0.001), with strong negative correlations to fasting, 1-hour and 2-hour postprandial glucose levels. Receiver operating characteristic (ROC) curve analysis revealed excellent diagnostic performance (area under the curve AUC: 0.95 95% CI: 0.91–0.99 for miR-326 and 0.96 95% CI: 0.93–0.99 for miR-532-3p), robustly confirmed by bootstrap resampling. Functional analyses linked these miRNAs to phosphoinositide 3-kinase (PI3K)/Akt and Rap1 signaling pathways. Conclusion This study provides compelling evidence for circulating miR-326 and miR-532-3p as a synergistic pair of biomarkers for GDM. Their high diagnostic accuracy and mechanistic insights into metabolic dysregulation position them as a promising complementary tool for early GDM risk assessment and for understanding GDM pathophysiology.

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