An Exploratory Biomarker Study of First-Trimester Circulating miRNAs Associated with Later Gestational Diabetes Mellitus

Gestational diabetes mellitus (GDM) develops silently during early pregnancy, yet its earliest circulating molecular signatures remain poorly defined. In this exploratory biomarker study, we characterized first-trimester circulating microRNA (miRNAs) associated with later GDM using a pool-based small RNA sequencing approach. Using a systematic and unbiased sequencing strategy with locus-level miRNA resolution, we profiled the first-trimester plasma miRNome and prioritized a set of 18 mature miRNAs from among 255 detected species. Set-level functional enrichment analyses based on curated and predicted miRNA-target interactions derived primarily from cellular and tissue-based studies showed annotation-based convergence on pathways related to Ca²⁺ homeostasis, glucagon-insulin regulatory circuits, and PI3K-AKT signaling. Network analysis indicated coordinated associations among these miRNAs and shared target pathways involved in insulin secretion and insulin sensitivity. Key contributors-including miR-29a-3p, miR-29c-3p, miR-146a-5p, let-7a-5p, and miR-182-5p-were linked, through in silico target annotation, to central metabolic regulators such as <i>PTEN</i>, <i>PIK3R1</i>, <i>AKT1</i>, <i>AKT2</i>, and components of Ca²⁺ signaling (<i>ATP2A2</i>, <i>CALM1/3</i>, <i>ITPR1</i>, <i>RYR2</i>). These circulating miRNAs should be interpreted primarily as biomarkers reflecting coordinated metabolic states rather than as direct causal mediators. Most identified miRNAs have not been previously reported in the context of first-trimester GDM, supporting the exploratory and hypothesis-generating nature of this circulating miRNA signature in early gestational metabolic research.