Abstract
The uterus is a remarkable organ in its ability to undergo extensive tissue damage during menstruation and parturition, yet achieves efficient, scar-free repair. Coordinated regulation of this regenerative process is essential for uterine homeostasis and fertility; however, the underlying mechanisms remain incompletely understood. Here, we demonstrate that mesenchymal-epithelial transition (MET) contributed to postpartum endometrial re-epithelialization using Pdgfrα (CreERT2/+) ; Rosa26-tdTomato (fl/+) lineage tracing mice. Flow cytometry revealed a marked increase in mesenchymal-derived (MD) epithelial cells during active tissue repair. Notably, these cells were transient, undergoing clearance primarily via apoptosis following completed epithelial restoration. We also identified a migratory population of transitional cells of mesenchymal origin within the mesometrial stroma that incorporated into the luminal epithelium, consistent with an active MET program. Single-nucleus RNA sequencing (snRNA-seq) revealed that MD epithelial cells exhibited gene expression profiles associated with cell adhesion and cytoskeletal remodeling, while transitional cells were enriched for genes involved in junctional assembly and actin dynamics. MET-associated genes were significantly upregulated in both transitional and MD epithelial populations. Cell-cell communication analysis highlighted WNT, BMP, and EPHA signaling as candidate regulators of MET during regeneration. Together, these findings provide confirmation of MET as a physiologic mechanism of postpartum endometrial epithelial repair and uncover a coordinated signaling network that facilitates this process. Perturbations in MET may contribute to pathologies such as endometriosis or endometrial cancer, underscoring the importance of understanding mesenchymal-epithelial plasticity in both normal and disease states. SIGNIFICANCE: The mammalian endometrium undergoes repeated injury and repair during menstruation (women) and pregnancy (most eutherians), yet exhibits a remarkable capacity for rapid, scar-free healing that mediates infection, inflammation and hemorrhage. Despite its clinical relevance, the molecular regulation of endometrial regeneration remains poorly defined. Using a transgenic lineage-tracing mouse model, we identified mesenchymal-derived (MD) epithelial and transitional cells during the regenerative window and revealed a critical role for mesenchymal-epithelial transition (MET) in this process. Single-nucleus RNA sequencing further uncovered functional characteristics of these cells and highlighted WNT, BMP, and EPHA signaling as potential regulators of MET. These findings provide new insight into the cellular and molecular framework of endometrial regeneration and have important implications for diseases involving aberrant tissue repair.