A hormonally regulated gating mechanism controls EMT timing to ensure progenitor cell specification occurs prior to epithelial breakdown

Abstract

Epithelial-to-mesenchymal transitions (EMTs) are essential for morphogenesis, converting static epithelial cells into migratory mesenchymal cells. As EMT disrupts epithelial barriers, its timing must be tightly regulated during development. We investigated this regulation in the Drosophila posterior embryonic midgut, where the GATA transcription factor Serpent (Srp) drives a collective partial EMT in the principal midgut epithelial cells (PMECs). Srp is expressed well before PMEC-EMT initiation and is also required for an earlier event: the specification and delamination of endoblasts, which give rise to different midgut lineages including adult midgut progenitors (AMPs). Here we identify the steroid hormone ecdysone as a temporal cue that triggers PMEC-EMT. Although Srp activates and represses numerous genes during early stages of midgut development, including polarity regulators such as crumbs (crb), the removal of polarity proteins and E-Cadherin from the membrane of PMECs only properly occurs in the presence of ecdysone signalling. This suggests that Srp primes cells transcriptionally for EMT, which proceeds only once polarity and adhesion proteins are downregulated post-translationally. When PMECs undergo a premature EMT, this leads to a reduction in the number of progenitor cells, suggesting that maintaining epithelial integrity until after endoblasts are selected is crucial for early fate decisions. These findings reveal that ecdysone functions as a EMT gatekeeper, ensuring that EMT is temporally restricted to occur only after progenitor specification, and in coordination with other developmental processes. More broadly, our work shows that in this context, transcriptional priming of EMT can be uncoupled from its execution, which is post-translationally triggered by hormonal signals. Given the conserved role of nuclear steroid hormone receptors as GATA co-factors and their association with poor prognosis in cancer, this may represent a broader strategy by which systemic cues regulate EMT in development and cancer.