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

Abstract

Background Gestational 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. Methods Discovery: 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. Results A 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. Conclusions During 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.

The implications of exosomes in pregnancy: emerging as new diagnostic markers and therapeutics targets

“…It has been found that CD63+ and HSP70+ exosomes are present in the uterine luminal fluid (ULF) of cyclic and pregnant sheep [ 91 ]. Exosomes are involved in the suitable interaction between the embryo and uterine endometrium required for successful embryo implantation during pregnancy [ 92 ]. Endometrial epithelium can release exosomes into the uterine cavity and transfer specific miRNAs to the blastocyst or endometrial epithelial cells to enhance embryo implantation [ 93 ].…”

Extracellular Vesicles—New Players in Cell-to-Cell Communication in Gestational Diabetes Mellitus

“…SIRT1-deficient mice demonstrate increased release of EVs from adipose tissue with a potential systemic effect on metabolic regulation [ 87 ] leading to impaired glucose tolerance and insulin sensitivity. It has been determined that GDM patients show differences in the expression of other proteins such as spectrin alpha erythrocytic (SPTA)-1, CAMK 2β, PAPP-A, Perilipin 4, fatty acid-binding protein (FABP) 4 and hexokinase-3 [ 17 ], and all proteins were previously shown to also be differentially expressed and related to insulin resistance [ 88 , 89 , 90 ]. Taken globally, these data suggest that EVs are involved in changes in insulin sensitivity by transferring bioactive molecules to target cells that might lead to GDM.…”

“…Recently, it has been identified that miRNAs, miR-16-2-3p, miR-16-5p, miR-1910-5p, miR-423-5p, miR-92a-3p and miR-92b-3p are differentially expressed in GDM compared to normal glucose tolerance (NGT) pregnancies [ 17 ]. Interestingly, the expression of miR-423-5p and miR-16-2-3p is reported to be similarly altered in type 2 diabetes [ 96 , 97 ].…”

“…To date there is a paucity of data defining changes in the release, role and diagnostic utility of circulating EVs in pregnancies complicated by GDM. The presence of placenta-derived EVs in maternal circulation have been characterised by the co expression of a placental marker (i.e., placental alkaline phosphatase (PLAP)) [ 12 , 14 ], which has allowed detection of changes in the levels and content of EVs in GDM [ 11 , 15 , 16 , 17 , 18 ]. Thus, the aim of this review is to discuss the current criteria of diagnosis of GDM and how EV research might have a potential role in the early identification of women at risk to develop GDM.…”

Small Non-Coding RNAs in the Human Placenta: Regulatory Roles and Clinical Utility

“…Increased presence of these nucleic acids in maternal serum have been observed in pregnancy complications, where expression of interferons and cytokines trigger cellular dysfunction resulting in their diffusion into maternal circulation ( Konečná et al, 2018 ). Several miRNAs originating from extracellular vesicles in maternal plasma were found to differ in expression by gestational age in women with and without gestational diabetes ( Nair et al, 2021 )—a condition wherein the increased insulin demands required during pregnancy aren’t met.…”