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.
INTRODUCTION:Globally, more than two million babies are stillborn every year. The primary cause of stillbirth is placental dysfunction, which impairs oxygen and nutrient delivery to the fetus. From a physical perspective, the efficiency of this exchange depends on placental blood flow and microvascular villous structure. Doppler ultrasound (US) imaging is used clinically to assess abnormal placental blood flow, but evaluation of placental structure remains largely observational. Texture analysis of B-mode US images offers a quantitative approach to indirectly characterize placental structure through analysis of US image pixels. Therefore, the objective of this study was to investigate whether integrated quantitative biomarkers derived from placental texture, Doppler ultrasound, and standard clinical variables improve the assessment of pregnancies associated with adverse fetal outcomes. METHODS:Patients (N = 129) were recruited, and B-mode US images and Doppler measurements were acquired at visit 1 (28-32 weeks of gestation) and visit 2 (34-38 weeks of gestation). Clinical outcome data were used to define patients as High-Risk based on previously established markers of significant stillbirth risk (e.g., preeclampsia with severe features, severe FGR, perinatal hypoxia). Logistic regression and random forest models were used to evaluate the predictive value of each modality with clinical variables and in combination. RESULTS:Quantitative placental texture features demonstrated predictive performance comparable to Doppler ultrasound measurements, supporting their associations with adverse fetal outcomes. Integrating clinical, Doppler, and texture features further improved model performance compared with models developed from clinical variables alone or paired with either imaging modality individually. These findings demonstrate that quantifying placental US texture provides complementary information to Doppler and clinical variables. While further validation in larger external cohorts is required, this work supports the potential of quantitative multimodal ultrasound biomarkers for assessing pregnancies with adverse fetal outcomes.
The pulsation of blood in the umbilical artery and the feto-placental circulation has been a focus for computational modelling because of the important association between the patterns of pulsation and placental pathology. This review examines different approaches that have been used to study this relationship and the insights that can be drawn from these computational models. Wave reflection in the umbilical artery is presented both to aid in the interpretation of these patterns and as an example of how modelling approaches can be used to unify disparate hemodynamic measurements. The linkage between modelling and measurement methodologies is examined along with the considerations for model selection when approaching a new research question.
INTRODUCTION:The small GTPase RAC1 is crucial for blastocyst attachment to the endometrium during embryo implantation. RAC1 is regulated by RHOH, whose role in pregnancy establishment we investigated. METHODS:We analyzed uterine RHOH expression by immunoblotting during the endometrial receptivity window for embryo attachment and miscarriage (LPS-, and IFN-γ-induced), and abortion (RU-486-, and ormeloxifene-induced), using a mouse model, human-origin placenta and decidua-placenta, and human endometrial epithelial cells. Additionally, we determined the cellular localization of RHOH by immunohistochemistry. RESULTS:RHOH levels were elevated in placentas from human miscarriage cases (n = 7) and in decidua-placenta tissues (n = 13) from these cases. In the mouse model, RHOH expression was downregulated at the implantation site during both receptive and post-receptive stages. Conversely, during pseudopregnancy, uterine RHOH expression was higher in the pre-receptive stage. RHOH was primarily localized in stromal and epithelial cells at all stages examined. In the decidualized uterus, RHOH expression was reduced and predominantly detected in stromal cells. Additionally, upregulation of RHOH was observed in the uterus from miscarriage/abortion cases and inter-implantation sites in non-miscarriage/abortion mouse models. Furthermore, the downstream effector RAC1 was also increased in the miscarriage/abortion mouse models. DISCUSSION:RHOH was upregulated in the placenta and decidua-placenta tissues of human miscarriage cases. Similarly, in mouse models, RHOH expression increased in the endometrium during miscarriage/abortion. In contrast, RHOH expression decreased at implantation sites and in the decidualized endometrium. These findings suggest that RHOH functions as a regulator of endometrial receptivity, potentially involving RAC1, and that their upregulation is associated with miscarriage/abortion.
BACKGROUND:Preterm premature rupture of membranes (PPROM) before full term can lead to adverse pregnancy outcomes. Chorioamnionitis is the main cause of premature birth. This investigation aims to evaluate the clinical value of miR-338-3p in PPROM with histological chorioamnionitis (HCA), and its mechanism in human amniotic epithelial cells (hAECs). METHODS:215 patients with PPROM were included (PPROM-HCA group (n = 105), PPROM group (n = 110)). MiR-338-3p and PTN expression in the tissues were measured via real time quantitative polymerase chain reaction. The diagnostic value of miR-338-3p for PPROM with HCA was evaluated using receiver operating characteristic analysis. Cell proliferation, apoptosis and inflammatory factor levels were quantified via cell counting kit-8, flow cytometry and enzyme-linked immunosorbent assay. RESULTS:Compared with PPROM group, miR-338-3p was upregulated in the tissues of PPROM-HCA patients, while PTN was downregulated. MiR-338-3p had a good diagnostic value for PPROM-HCA patients. MiR-338-3p overexpression suppressed hAECs proliferation, promoted apoptosis, and increased interleukin-6 and tumor necrosis factor-α levels; inhibition of miR-338-3p showed the reverse phenotypes. PTN was the target gene of miR-338-3p. PTN overexpression partially reversed the cell proliferation inhibition, increased apoptosis and release of inflammatory factors induced by the upregulation of miR-338-3p. CONCLUSION:MiR-338-3p is upregulated in PPROM-HCA patients, and it has a good diagnostic value. MiR-338-3p modulates the proliferation, apoptosis and inflammatory response of hAECs via targeting PTN. These findings offer novel insights into the pathological research of PPROM-HCA.
BACKGROUND:Maternal obesity is a well-established risk factor for preeclampsia; however, whether increased maternal BMI is associated with biologically distinct subtypes of this disorder remains unknown. OBJECTIVE:To investigate whether maternal BMI is associated with differences in placental gene expression patterns observed in preeclampsia and to identify transcriptomic signatures with potential relevance for future biomarker research. STUDY DESIGN:This exploratory analysis used placental transcriptomic data obtained from the Gene Expression Omnibus. Differential expression within prespecified contrasts encoding preeclampsia status and maternal BMI was analyzed using a multivariable linear modeling framework with adjustment for relevant confounders. In a secondary analysis, maternal BMI was modeled continuously with an interaction term between BMI and preeclampsia status. RESULTS:Placental transcriptomic data from 132 placentas representing a range of maternal hypertensive and normotensive states were analyzed. Genes commonly associated with preeclampsia, including FLT1, LEP, HTRA4, and FSTL3, were differentially expressed across all BMI categories. SERPINA3 demonstrated the strongest differential expression in the obese contrast, with an estimated eight-fold increase (95% CI: 3.9-16.6) in preeclamptic pregnancies complicated by obesity, whereas expression differences were smaller and not statistically significant in healthy-weight and overweight contrasts. Gene set enrichment analysis suggested heterogeneity in the underlying biology of preeclampsia across BMI strata. CONCLUSION:Placental transcriptional patterns associated with preeclampsia differed across maternal BMI strata. These findings are consistent with previously proposed molecular subtypes of preeclampsia and highlight SERPINA3 as a candidate transcriptomic marker warranting further investigation, specifically in pregnancies complicated by obesity.
OBJECTIVES:The primary objective was to elucidate whether placental histopathology representative of intrauterine insults during pulmonary vascular development are associated with a diagnosis of persistent pulmonary hypertension of the newborn (PPHN). The secondary objective was to compare placental histopathologic lesions across PPHN etiologies. STUDY DESIGN:We conducted a case-control study of mother-infant dyads ≥35 weeks gestation who delivered at a tertiary care center between 2020 and 2025. Cases were infants diagnosed with PPHN and treated with inhaled nitric oxide; unaffected controls were infants without congenital anomalies. Placentas underwent blinded histopathologic review using standardized criteria. Multivariate logistic regression modeling was used to control for confounding maternal and infant factors. RESULTS:106 placentas were analyzed (53 PPHN, 53 controls). Placental lesions were significantly more common in PPHN placentas, including fetal vascular malperfusion (30.2% vs 9.4%, p < 0.01), placental inflammation (66.0% vs 37.7%, p < 0.01), chronic presence of meconium (43.4% vs 15.2%, p < 0.01), and chorangiosis (7.6% vs 0%, p = 0.04). In adjusted analyses compared to controls, among 41 placentas with fetal development etiologies (e.g. congenital anomalies) of PPHN, fetal vascular malperfusion, placental inflammation and fetal inflammatory response were more common. Among 12 placentas with typical causes of PPHN (e.g. meconium aspiration syndrome), placental inflammation, maternal and fetal inflammatory responses, and meconium were more common. CONCLUSIONS:PPHN placentas demonstrate lesions of malperfusion, inflammation, and chronic meconium, suggesting a complex interplay between intrauterine hypoxia and inflammation as a potential mechanism for the abnormal pulmonary vascular development and function seen in PPHN.
INTRODUCTION:The haemodynamics of maternal blood flow in the intervillous space (IVS) impacts placental exchange efficiency. The resulting shear stress can also affect syncytiotrophoblast function, and in turn placental development. Here, we use anatomically-informed multiscale modelling approaches to predict flow in the IVS and syncytiotrophoblast shear stress in fetal growth restriction (FGR). METHODS:Three-dimensional placentone models were established and parameterised to normal and FGR scenarios. Normal term and FGR placental tissue punches were microCT imaged, segmented, and used in tissue-level computational fluid dynamics simulations to predict syncytiotrophoblast shear stress. Tissue and placentone-level models were combined to predict syncytiotrophoblast shear stress ranges. Mechanosensing protein expression was determined by immunohistochemistry. RESULTS:Placentone-level models demonstrate variation in IVS flow velocity across the placental depth, with higher velocity regions at spiral artery mouths that extended further in FGR. Tissue-level models predicted higher levels of syncytiotrophoblast shear stress in FGR. Combined models predicted a greater proportion of tissue is exposed to higher shear stresses in FGR, with ∼2-fold increase in peak shear stress proximal to spiral artery openings. Including septal veins increased penetration of maternal blood and the proportion of tissue exposed to higher shear. In FGR, the syncytiotrophoblast had greater Dynein-1 and lower Kinesin-2 and TRVP6 expression, but no difference in IFT88, Polycystin-2 or Piezo-1 expression. CONCLUSIONS:In FGR, impaired remodelling of the spiral arteries and changes in villous tissue architecture combine to increase the proportion of placental tissue exposed to higher shear stress. Septal veins and central cavities act synergistically to ensure adequate placental perfusion.
INTRODUCTION:The monoterpenoid eucalyptol (EUC; 1,8-cineole) has a vasodilatory effect on rat arteries; however, its effects on the human fetoplacental vasculature had not yet been described. Thus, the objective of this study is to evaluate the endothelium-independent vasorelaxant effect of EUC on human umbilical veins from normoglycemic parturients (HUV-NG) and parturients with gestational diabetes mellitus (HUV-GDM). METHOD:Denuded HUV rings were mounted in organ bath for isometric recordings under different experimental protocols in the presence of EUC (1-3000 μM). RESULTS:EUC (1-3000 μM) inhibited vasoconstriction induced by KCl (60 mM) and 5-HT (10 μM), exhibiting greater potency in HUV-GDM. EUC (3000 μM) prevented vasoconstriction induced by CaCl2 or BaCl2 (0.1-30 mM) in HUV-NG. However, its efficacy in preventing vasoconstriction induced by high concentrations of BaCl2 was attenuated in HUV-GDM. The presence of potassium channel blockers (TEA, 4-AP, GLI, and BaCl2) reduced the vasorelaxant efficacy and potency of EUC, differentially in HUV-NG and HUV-GDM. DISCUSSION:The results suggest that EUC acts by modulating ion channels to induce vasorelaxation in HUV, possibly through the inhibition of voltage-operated calcium channels and the activation of potassium channels in vascular smooth muscle. Furthermore, the pathophysiological changes induced by GDM in HUV appear to alter the pharmacodynamics of EUC; nevertheless, the vasorelaxant effect of EUC was more potent via electromechanical and pharmacomechanical pathways in HUV-GDM. Thus, the current study provides a basis for the development of new pharmacological vasodilator strategies.
BACKGROUND:Placental malaria contributes to adverse birth outcomes, preterm birth, intrauterine growth restriction, and low birth weight, yet the early molecular responses of the human placenta to Plasmodium falciparum remain poorly defined. The aim of this study was to explore the early transcriptional responses of human placental explants to P. falciparum infection using an in vitro co-culture model. METHODS:Human placental explants were co-cultured with P. falciparum-infected erythrocytes. Tissue responses were evaluated through histopathology, LDH release assays, and transcriptomic profiling. RNA was extracted for downstream gene expression analysis, and differentially expressed genes were subjected to functional enrichment analysis. RESULTS:Short-term exposure induced evident syncytiotrophoblast injury and increased LDH release. Transcriptomic profiling identified 197 differentially expressed genes, predominantly upregulated, involving pathways related to tissue remodeling, epithelial organization, angiogenesis, and metabolic regulation. Notably, several upregulated genes overlapped with transcriptional programs associated with the placenta-brain axis, including genes involved in neurotransmitter synthesis, calcium signaling, and synaptic organization. Downregulation of LPL suggested early disruption of placental lipid processing. CONCLUSION:P. falciparum triggers structural and transcriptional remodeling in human placental tissue, affecting pathways essential for epithelial integrity, nutrient metabolism, and endocrine-neuroactive signaling. These molecular signatures provide a potential link between placental infection and long-term nutritional and neurodevelopmental outcomes observed in malaria-exposed children.
INTRODUCTION:Preeclampsia (PE) is one of the leading causes of maternal and infant mortality. Increase of soluble fms-like tyrosine kinase-1 (sFLT-1) is thought to be a key player in pathophysiology. Since preclinical studies showed therapeutical effects of esomeprazole (Eso), we hypothesized treatment may target sFLT-1 in PE. METHODS:Using the PE/FGR (fetal growth restriction) model with systemic human sFLT-1 overexpression, mice were treated with Eso or vehicle. We analyzed dams, fetal and placental tissues regarding sFLT-1 at day 18.5 dpc and determined hypoxia of placentas using the marker pimonidazole. RESULTS:Eso led to a trend of decrease in sFLT-1 serum levels. As shown previously by us sFLT-1 increase resulted in different severity levels of FGR. Exclusive maternal sFLT-1 overexpression (PE wt) fetuses revealed a mild reduction in weights, combined maternal and feto-placental sFLT-1 (PE het) led to strong reductions in weights and non-viability of the fetuses. Upon Eso an improvement in fetal outcome shown by an increased rate of fetal viability and a significant increase in weight of PE het fetuses was observed. Furthermore, upon Eso sFLT-1 (sFLT-i13) mRNA expression significantly decreased in the mesometrial triangle tissue (MT) and in placentas of PE het fetuses and maternal kidneys. A significant decrease of pimonidazole in MT of PE wt fetuses and a trend of decrease in PE het fetuses upon Eso showed less hypoxia in the maternal compartment. DISCUSSION:These results could indicate a potential cytoprotective effect of Eso and an improvement of the hypoxic conditions at the maternal-fetal interface during PE.
Background Sepsis during pregnancy is associated with increased risks of fetal growth restriction, preeclampsia, and placental abruption even after maternal recovery. Characterizing placental pathology after antepartum sepsis may clarify mechanisms linking infection to subsequent pregnancy complications. Methods We performed placental histopathologic analysis on individuals delivering at a single institution. Cases had resolved antepartum sepsis (defined as clinical recovery ≥7 days before delivery), and controls were individuals without an antepartum infection hospitalization. Exclusion criteria included multifetal gestation, fetal anomalies, TORCH infections, and delivery <34 weeks' gestation (chosen to minimize gestational age-dependent histologic variability). All placentas were reviewed by a subspecialty placental pathologist and lesions characterized according to the Amsterdam criteria. Clinical characteristics and histopathologic findings were compared using Fisher's exact and Wilcoxon rank-sum tests. Results Twenty-two sepsis-exposed pregnancies and 49 controls were included. The most common infection sources were urinary tract (36.4%) and respiratory (36.4%), diagnosed at a mean (SD) gestational age of 25.3 (9.2) weeks. The sepsis group was younger (31.6 [4.6] vs 34.6 [4.7] years; p = 0.017) and had higher rates of tobacco use (13.6% vs 0%; p = 0.027) and pregestational diabetes (13.6% vs 0%; p = 0.027). Sepsis-exposed placentas had significantly higher rates of maternal vascular malperfusion (31.8% vs 0%; p < 0.001), placental infarction (13.6% vs 0%; p = 0.027), avascular villi (27.3% vs 4.1%; p = 0.009), histologic chorioamnionitis (50.0% vs 17.0%; p = 0.015), and meconium-staining (50.0% vs 14.3%; p = 0.003). Conclusions Resolved antepartum sepsis is associated with increased placental vascular and inflammatory pathology, suggesting lasting placental injury after severe systemic infection that may underlie adverse pregnancy outcomes.
INTRODUCTION:Diabetes in pregnancy (DIP) is associated with adverse maternal and fetal outcomes. Placental iron homeostasis is tightly regulated to ensure adequate fetal supply while preventing oxidative injury. Dysregulated iron metabolism may promote ferroptosis. Altered placental iron homeostasis in DIP may reduce the Placental Iron Deficiency Index, thereby compromising fetal iron endowment. The study evaluated alterations in placental iron homeostasis and ferroptosis-related pathways in DIP compared with healthy pregnancies (HP). METHODS:In this cross-sectional analytical study, 120 pregnant women were recruited and divided into HP (n = 60) and DIP (n = 60) groups. Sixteen placentae from each group, at ≥34 weeks of gestation, were analysed. Placental expressions of hepcidin, ferritin, FPN1, TfR1, TGF-β, NQO1, and GPX4 were assessed by qPCR, western blotting and IHC. Further, the perinatal outcomes were measured in each group. RESULTS:Placental gene expression of hepcidin was increased in DIP compared with HP, accompanied by reduced ferritin and TfR1 expression. Protein analyses confirmed increased hepcidin and decreased levels of ferritin, TfR1, and FPN1. Immunohistochemistry localised these alterations predominantly to the syncytiotrophoblast. TGF-β and NQO1 transcripts were elevated, whereas GPX4 protein expression was reduced in DIP placentae. The newborns of DIP had an adverse perinatal outcome compared to HP. DISCUSSION:The DIP is associated with profound disruption of placental iron homeostasis and impaired ferroptosis defence mechanisms. Upregulation of hepcidin, alongside suppression of key iron transport proteins and reduced GPX4, suggests a pro-oxidative, iron-dysregulated placental environment. These molecular alterations may contribute to trophoblast dysfunction and adverse pregnancy outcomes, highlighting potential therapeutic targets.
INTRODUCTION:Placental stiffness measured using shear wave elastography has been proposed as a potential indicator of placental structural characteristics. However, the measurement sites for placental stiffness have not yet been standardized. In this study, we aimed to compare the placental stiffness between extrachorial (circumvallate and circummarginate) and normal placentas to evaluate the impact of measurement site selection on the assessment. METHODS:In this cross-sectional study of 169 singleton placentas (135 normal, 34 extrachorial) delivered after 32 weeks, the shear wave velocity (SWV) was measured at the placental marginal area and near the umbilical cord insertion site. A histopathological evaluation was performed in representative cases. RESULTS:The median SWV at the marginal area was significantly higher in the extrachorial placenta group than in the normal placenta group (2.20 vs. 2.07 m/s, p < 0.01). By contrast, no significant difference was observed near the umbilical cord insertion site between the groups. Secondary analyses showed consistent findings for the circumvallate and circummarginate subtypes. The histopathological examination revealed marked fibrin deposition on the fetal side at the placental margin in extrachorial placentas, which may contribute to localized increases in placental stiffness. DISCUSSION:These findings suggest that placental stiffness measurements may be influenced by the choice of measurement site, particularly in the presence of localized structural changes such as those observed in extrachorial placentas. Measurements near the umbilical cord insertion site may be less affected by marginal structural variations and may better reflect the overall mechanical properties of the placenta.
BACKGROUND:Premature rupture of membranes (PROM) is a major cause of preterm birth and perinatal morbidity. Disruption of fetal membrane integrity is central to PROM pathogenesis; however, the molecular mechanisms underlying membrane weakening remain incompletely understood. METHODS:Proteomic analysis was performed on fetal membrane tissues from patients with early PPROM, late PPROM, full-term PROM, and full-term normal pregnancies. An LPS-induced infection-associated preterm birth mouse model was established to evaluate fetal membrane pathology, hormone levels, inflammatory responses, and Kisspeptin-1 (KISS1) expression. Human amniotic epithelial cells (hAECs) were used for in vitro studies to investigate the regulatory relationship between progesterone and KISS1 and their effects on cell viability, apoptosis, migration, and adhesion. RESULTS:Proteomic profiling revealed significant upregulation of KISS1 in PROM, particularly in PPROM, accompanied by downregulation of adhesion-related proteins and enrichment of steroid metabolic pathways. In PROM mice, fetal membranes exhibited structural disruption, reduced progesterone levels, elevated inflammatory cytokines, and increased KISS1 expression. In hAECs, progesterone treatment suppressed KISS1 expression, whereas inhibition of progesterone signaling upregulated KISS1. Conversely, KISS1 overexpression reduced progesterone secretion, while KISS1 knockdown increased progesterone levels, indicating a reciprocal regulatory relationship. Progesterone inhibition impaired hAEC viability, enhanced apoptosis, reduced migration, and decreased expression of adhesion proteins FN1 and VTN. These effects were partially reversed by KISS1 knockdown. CONCLUSIONS:The progesterone-KISS1 regulatory axis plays a critical role in maintaining amniotic epithelial cell function and fetal membrane integrity. Dysregulation of this axis promotes cellular dysfunction and contributes to PROM, highlighting KISS1 as a potential biomarker and therapeutic target.
Background Preeclampsia (PE) causes high maternal and perinatal morbidity/mortality, with unclear pathogenesis. Given the emerging roles of cuproptosis and M1 macrophage polarization in PE progression, this study investigated the cuproptosis- and M1 macrophage-associated transcriptomic network to identify specific molecular targets underlying trophoblast damage. Methods Placental transcriptomes of controls and severe PE patients were analyzed. Immune infiltration, WGCNA (cuproptosis/M1 macrophage-related modules), DEG screening, and machine learning identified hub genes. Validation was done in GSE148241, H2O2-induced HTR-8/SVneo cells, and PE placental tissues. GYS2/NRIP1 functions were tested via rescue assays; their association was verified by ChIP and dual-luciferase assays. Results Severe PE had increased placental M1 infiltration. WGCNA's blue module correlated with cuproptosis/M1 macrophages, and 5 hub genes were identified, with GYS2 downregulated consistently. GYS2 overexpression attenuated H2O2-induced oxidative stress, cuproptosis, and M1 polarization. NRIP1 inhibited GYS2, and NRIP1 knockdown protected trophoblasts via GYS2 upregulation. Conclusion NRIP1 knockdown-induced GYS2 upregulation alleviates H2O2-induced damage in trophoblasts; GYS2 is a potential PE therapeutic target.
