Abstract: Adenomyosis is a challenging gynecological condition to study due to the absence of in vitro models that accurately replicate endometrial tissue dynamics across the menstrual cycle. In this study, we established an assembloid model that simulates cycle-dependent endometrial responses and mirrors the cellular and molecular features of adenomyosis, including lesion-specific epithelial and stromal heterogeneity. Single-cell transcriptomic revealed that ectopic epithelial cells exhibit a luminal-dominant and glandular-deficient profile during the secretory-like phase following minimal differentiation medium (MDM) treatment. These changes were linked to MDM-induced stromal shifts, characterized by the loss of BMP4+ stromal cells and an increase in CRYAB+IL15+ stromal cells, disrupting stromal-epithelial BMP signaling and enhancing WNT signaling pathways. Additionally, ectopic epithelial and stromal cells demonstrated increased immunity and angiogenesis activities. This assembloid model provides a robust platform for studying adenomyosis pathogenesis and highlights WNT signaling as a potential therapeutic target, paving the way for more targeted and effective treatments.
Funding Source: National Key Research and Development Program of China (2022YFC2702200, 2023YFA1801800, 2023YFA1800300), NSFC (82471684, 32330030, 32270840, 32270908), Science and Technology Commission of Shanghai Municipality 23JC1403700
