Introduction Gestational diabetes mellitus (GDM) is a major public health challenge characterized by placental immunometabolic dysregulation. This study aimed to identify molecular signatures associated with GDM-related placental pathology and evaluate their diagnostic and therapeutic implications. Methods Two bulk placental transcriptomic datasets (GSE70493 and GSE263483) and one single-cell dataset (GSE173193) were analyzed. Differential expression analysis, WGCNA, and machine-learning feature selection (LASSO and SVM-RFE) were used to identify candidate gene signatures. A two-gene logistic regression model was constructed and externally validated. Immune-cell correlation rewiring and single-cell in silicoperturbation analysis explored immune-network alterations and regulatory roles of identified genes. A drug-prioritization framework was used for therapeutic screening. Results Intersecting WGCNA modules with differentially expressed genes identified 29 candidates enriched in antigen presentation and mononuclear phagocyte functions. Dual-algorithm selection converged on HLA-DQA2 and FGL2. The two-gene model achieved an AUC of 0.786 in the discovery cohort and 0.813 in the external validation cohort. Immune rewiring analysis indicated a shift toward M1 macrophage polarization. Single-cell perturbation analysis supported regulatory roles of HLA-DQA2 and FGL2 within the macrophage lineage. Drug analysis reaffirmed insulin, metformin, and glyburide as stable therapeutic anchors. Discussion The HLA-DQA2/FGL2 signature captures key aspects of placental immunometabolic dysregulation in GDM, including altered antigen presentation and macrophage-associated immune rewiring. This integrated framework provides candidate biomarkers for GDM risk stratification and generates computational hypotheses for future mechanistic and therapeutic studies.
OBJECTIVES:Prenatal diagnosis of the placenta accreta spectrum (PAS) relies on subjective anatomical MRI interpretation and lacks objective quantitative biomarkers. This study aimed to validate diffusion MRI myometrial tractography for PAS detection and compare its performance with conventional visual MRI diagnosis. MATERIALS AND METHODS:This prospective study enrolled 154 pregnant women with suspected PAS. All subjects underwent routine MRI and DTI. Tractography-derived FA, fiber density and ADC were measured. Diagnosis was confirmed by 2023 FIGO criteria; participants were stratified into a 70% training set and independent 30% test set. Statistical analyses included group comparisons, ROC analyses with DeLong's test, and ICC for interobserver reliability. RESULTS:PAS patients exhibited lower FA (p < 0.001), reduced fiber density (p < 0.001) and higher ADC (p = 0.044). The clinically adjusted FA + fiber density model reached an AUC of 0.86 in the training set (conventional MRI AUC = 0.81) and maintained an AUC of 0.82 in the test set, comparable to conventional MRI. Combined parameters yielded an AUC of 0.82 for differentiating PAS grade 1 and grade 2. ICCs (0.788-0.948) indicated excellent interobserver agreement. CONCLUSION:Quantitative diffusion MRI tractography provides objective biomarkers for prenatal PAS evaluation. The combined FA and fiber density model yields stable diagnostic performance and delivers complementary quantitative information to mitigate subjectivity in conventional anatomical-MRI assessment. This proof-of-concept work requires further external multicenter validation before clinical adoption.
INTRODUCTION:Abnormal umbilical cord insertion may influence the risk of adverse outcomes in monochorionic twin pregnancies. This study aimed to evaluate the diagnostic accuracy of prenatal ultrasound in identifying different types of umbilical cord insertion across pregnancy trimesters. METHODS:This prospective monocentric study was conducted at Fondazione Policlinico Universitario Agostino Gemelli IRCCS, Rome, between 2021 and 2025. Monochorionic twin pregnancies with prenatal ultrasound assessment, obstetric outcomes, and postpartum placental examination were included. Umbilical cord insertion was assessed during each trimester, using placental examination as the reference standard. Sonographic classification included eccentric, marginal, and velamentous insertions, and proximate cord insertion (pPCI), defined as insertions ≤4 cm apart. Diagnostic performance was evaluated through sensitivity, specificity, predictive values, likelihood ratios, and accuracy. RESULTS:Among 184 monochorionic twin pregnancies, placental examination identified eccentric, marginal, and velamentous insertions in 53.0%, 35.0%, and 12.0% of cases, respectively. pPCI occurred in 9.3% (17/182). Ultrasound showed a sensitivity of 0.79, specificity of 0.93, and overall accuracy of 0.87 (95% CI 0.83-0.90). Accuracy improved from 60.0% in the first trimester to 80.9% and 77.5% in the second and third trimesters. Detection of pPCI showed 99% accuracy. Posterior placental location was associated with reduced diagnostic accuracy (OR 0.24; 95% CI 0.12-0.43; p < 0.001). DISCUSSION:Prenatal ultrasound reliably identifies umbilical cord insertion abnormalities in monochorionic twin pregnancies, with optimal performance especially in the second trimester, supporting its routine antenatal assessment.
BACKGROUND:Maternal high-fat diet (HFD) contributes to developmental programming through placental adaptations; however, the effects of exposure timing before and during pregnancy remain unclear. OBJECTIVE:To investigate the distinct effects of pre-pregnancy and gestational HFD on maternal metabolism, pregnancy outcomes, placental oxidative stress, placental gene-expression adaptations, and gut microbiota in rats. METHODS:Female Sprague-Dawley rats were fed a control diet (V) or HFD (H) before and/or during pregnancy, generating four groups (V/V, V/H, H/V, H/H). Maternal metabolic parameters, placental gene expression, oxidative stress, and gut microbiota composition were analyzed at gestational day 21. RESULTS:Pre-pregnancy high-fat diet increased maternal body and liver weights and elevated placental oxidative stress, whereas gestational high-fat diet increased adiposity and impaired glucose tolerance and insulin sensitivity without affecting pregnancy outcomes. Placental nutrient transporters, including glucose transporter 1 (Glut1) and solute carrier family 38 (Slc38) members, were mainly regulated by pre-pregnancy diet. Insulin-like growth factor 2 (Igf2) was influenced by gestational diet and its interaction with pre-pregnancy diet, while insulin-like growth factor 2 receptor (Igf2R) and insulin 2 (Ins2) expression were primarily influenced by pre-pregnancy diet. Forkhead box O1 (FoxO1) and Forkhead box O3 (FoxO3) exhibited differential responses to maternal dietary exposure. Gestational HFD also altered gut microbiota composition without affecting microbial diversity. CONCLUSION:Pre-pregnancy and gestational HFD exert distinct but complementary effects on maternal metabolism and placental adaptations. These findings highlight the stage-specific impact of maternal nutritional status on developmental programming and placental function.
Introduction The human placental trophoblast layer is a crucial barrier to the transfer of xenobiotics from maternal to fetal fluids. Intercellular spaces between adjacent trophoblast cells are sealed by bicellular and tricellular tight junctions (TJs), which respectively include claudins and angulins. However, which TJ components are involved in the barrier function of placental trophoblast cells remains unclear. Methods In this study, we used proteins that bind to the extracellular domain of claudins or angulins (hereafter referred to as binders) to investigate the contributions of various TJ components to barrier function in JEG-3 cells, a human trophoblast model. Results In this system, claudin binders enhanced the permeation of paracellular passage markers (dextrans with molecular mass of ≤150 kDa) and thus more effectively attenuated TJ integrity than did angulin binders. Gene expression analysis revealed that claudin-4 and angulin-1 are key components in bicellular and tricellular TJs, respectively. Conclusions This study is the first to characterize the barrier function of TJ components in JEG-3 cell monolayers, a human trophoblast model.
