Extracellular vesicle-associated miRNAs are an adaptive response to gestational diabetes mellitus

Soumyalekshmi Nair,Nanthini Jayabalan,Jezid Miranda,Gregory Duncombe,Fátima Crispi,Carlos Palma,Eduard Gratacós,Valeska Ormazabal,Martha Lappas,Elizabeth A Mccarthy,Dominic Guanzon,Priyakshi Kalita De Croft,Alexis Shub,Katherin Scholz-Romero,Gregory Rice,Emilio Diaz,Carlos Salomon,Andrew Lai,H David Mcintyre

doi:10.1186/s12967-021-02999-9

Abstract

BackgroundGestational diabetes mellitus (GDM) is a serious public health issue affecting 9–15% of all pregnancies worldwide. Recently, it has been suggested that extracellular vesicles (EVs) play a role throughout gestation, including mediating a placental response to hyperglycaemia. Here, we investigated the EV-associated miRNA profile across gestation in GDM, assessed their utility in developing accurate, multivariate classification models, and determined the signaling pathways in skeletal muscle proteome associated with the changes in the EV miRNA profile.MethodsDiscovery: A retrospective, case–control study design was used to identify EV-associated miRNAs that vary across pregnancy and clinical status (i.e. GDM or Normal Glucose Tolerance, NGT). EVs were isolated from maternal plasma obtained at early, mid and late gestation (n = 29) and small RNA sequencing was performed. Validation: A longitudinal study design was used to quantify expression of selected miRNAs. EV miRNAs were quantified by real-time PCR (cases = 8, control = 14, samples at three times during pregnancy) and their individual and combined classification efficiencies were evaluated. Quantitative, data-independent acquisition mass spectrometry was use to establish the protein profile in skeletal muscle biopsies from normal and GDM.ResultsA total of 2822 miRNAs were analyzed using a small RNA library, and a total of 563 miRNAs that significantly changed (p < 0.05) across gestation and 101 miRNAs were significantly changed between NGT and GDM. Analysis of the miRNA changes in NGT and GDM separately identified a total of 256 (NGT-group), and 302 (GDM-group) miRNAs that change across gestation. A multivariate classification model was developed, based on the quantitative expression of EV-associated miRNAs, and the accuracy to correctly assign samples was > 90%. We identified a set of proteins in skeletal muscle biopsies from women with GDM associated with JAK-STAT signaling which could be targeted by the miRNA-92a-3p within circulating EVs. Interestingly, overexpression of miRNA-92a-3p in primary skeletal muscle cells increase insulin-stimulated glucose uptake.ConclusionsDuring early pregnancy, differently-expressed, EV-associated miRNAs may be of clinical utility in identifying presymptomatic women who will subsequently develop GDM later in gestation. We suggest that miRNA-92a-3p within EVs might be a protected mechanism to increase skeletal muscle insulin sensitivity in GDM.

Highlights

Gestational diabetes mellitus (GDM) is a serious public health issue affecting 9–15% of all pregnancies worldwide
We identified a total of 9 genes (Suppressor of cytokine signaling 2 (SOCS2), Mothers against decapentaplegic homolog 1 (SMAD1), Upstream stimulatory factor 1 (USF1), Cyclin-dependent kinase inhibitor 1 (CDKN1A), TNF receptor superfamily member 6 (FAS), Suppressor of cytokine signaling 4 (SOCS4), Nitric oxide synthase, inducible (NOS2), and Janus kinase 2 (JAK2)) significantly differentially expressed in cells transfected with miR-92a-3p compared to negative miRNA (p < 0.05) (Fig. 6D). miR-92a-3p increased the expression of SOCS4 (1.2-fold), FAS (1.4-fold), Cyclin dependent kinase inhibitor 1A (CDKN1A) (1.5-fold), SMAD Family Member 1 (SMAD1) (1.3-fold), SOCS2 (2.2-fold), JAK2 (1.4-fold), USF1 (1.2fold), and Interferon gamma receptor 1 (IFNGR1) (1.4-fold)
As difference in the miRNA content within extracellular vesicles (EVs) at early pregnancy were observed, we developed a classification model using the expression of the miRNAs hsa-miR-let7i-5p, hsa-miR-10a-5p, hsa-miR-151b, hsamiR-16–2-3p, hsa-miR-16-5p, hsa-miR-1910-5p, hsamiR-423-5p, hsa-miR-92a-3p, and hsa-miR-92b-3p at early gestation, which can predict the risk of developing GDM later in pregnancy, with a classification efficiency of 100% and 82% for without and with cross validation, respectively

Summary

Introduction

Gestational diabetes mellitus (GDM) is a serious public health issue affecting 9–15% of all pregnancies worldwide. Gestational Diabetes Mellitus (GDM) is defined as glucose intolerance with onset or first recognition during pregnancy [1]. With a worldwide prevalence of 9–15%, GDM has been recognized as the most common medical complication during pregnancy [2]. Women with a history of GDM have increased risk of developing type 2 diabetes and cardiovascular disease, whilst their offspring have increased risks for early onset type 2 diabetes and obesity [4]. Our studies involving extracellular vesicles (EVs) have shown that they may have a key role in GDM [5,6,7]

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