AUTHOR=Lee Elaine , Kazemi Parinaz , Shafiei Shiva , Yull Sarah , Rana Mansuba , Tawil Nadim , Montermini Laura , Rak Janusz , Dufort Daniel 

TITLE=Extracellular vesicles as prospective biological indicators for midgestational placental complications in the mouse

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=Volume 13 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2025.1636335

DOI=10.3389/fcell.2025.1636335

ISSN=2296-634X

ABSTRACT=BackgroundPlacental dysfunction is often associated with reproductive complications such as preeclampsia, intrauterine growth restriction (IUGR), and preterm birth. Currently, the early diagnosis and intervention of these pathologies remain challenging due to the invasive nature of placental tissue sampling. Liquid biopsies of extracellular vesicles (EVs) released from the placenta have emerged as a prospective minimally invasive diagnostic strategy that could provide insight into the maternal-fetal interface because of the active role EVs play in mediating placental development and function. However, the lack of information on EVs directly from placenta at disease onset has questioned the representativeness of placental EVs as pathological indicators. To address these concerns, this study assessed the accuracy with which tissue-derived D10.5 placental EVs could identify phenotypes exhibited by a reproductively challenged Nodal conditional knockout mouse model at mid-gestation.MethodImplantation sites from female mice with a uterine-specific knockdown of the Nodal gene were examined from D8.5 to D14.5 utilizing histological analysis, Western blotting, and RT-qPCR to characterize their mid-gestational phenotypes. Placental EVs were then isolated from D10.5 placenta using enzymatic digestion, differential centrifugation, filtration, and size-exclusion chromatography. The final EV fractions were concentrated and validated with size analysis, canonical protein markers, and morphology assessment. Differential expression analysis across the EV samples was performed using proteomics and miRNA-Seq. Functional enrichment analysis of dysregulated EV factors was then completed using several gene ontology databases along with a literature review to determine whether placental EVs could indicate the reproductive abnormalities presented by the mutant mice.ResultsUterine-specific deletion of Nodal resulted in IUGR and fetal loss in mutant dams. Decidualization and placentation defects were observed, including thinner decidual and placental tissues, impaired angiogenesis, and an altered junctional zone within the maternal-fetal interface. Bioinformatics analysis of EV cargo identified 31 differentially expressed proteins and 10 miRNAs specifically linked to placental development, oxidative stress, angiogenesis, and immunomodulation. Notably, 15 of these proteins and six of these miRNAs have been previously associated with pregnancy complications, further supporting the prospects of placental EVs as biomarkers for various placental diseases.ConclusionThese findings suggest that placental EVs can reflect compromised placental function and could serve as pathological indicators for the early detection of pregnancy complications. Their potential diagnostic utility could improve maternal and neonatal health outcomes by enabling earlier intervention and monitoring of high-risk pregnancies.