Identifying Molecular Signatures of Distinct Modes of Collective Migration in Response to the Microenvironment Using Three-Dimensional Breast Cancer Models

Collective cell migration is a key feature of transition ductal carcinoma in situ (DCIS) to invasive (IDC) among many other cancers, yet the microenvironmental factors and underlying mechanisms that trigger collective remain poorly understood. Here, we investigated two factors, tumor-intrinsic hypoxia tumor-secreted (secretome), as triggers using three-dimensional (3D) discrete-sized microtumor models recapitulate hallmarks DCIS-IDC transition. Interestingly, induced distinct modes migration: directional radial 3D microtumors generated from same breast cancer line model, T47D. Without external stimulus, large (600 µm) T47D exhibited migration, while small (150 µm), non-hypoxic only when exposed secretome microtumors. To investigate hypoxia- secretome-induced vs. modes, performed differential gene expression analysis migratory compared with non-hypoxic, non-migratory controls. We propose unique signature sets related hypoxia, hypoxia-induced epithelial-mesenchymal (EMT), well migration. Gene Set Enrichment Analysis (GSEA) protein-protein interaction (PPI) network revealed enrichment potential between EMT, signatures for In contrast, EMT were not enriched suggesting complete may be required Survival identified genes associated low survival rate poor prognosis TCGA-breast dataset our (CXCR4, FOXO3, LDH, NDRG1), (EFEMP2, MGP), (MAP3K3, PI3K3R3). NOS3 was common signature. ATM, KCNMA1 (hypoxia signature), KLF4, IFITM1, EFNA1, TGFBR1 (migration signature) rate. conclusion, cultures controlled microenvironments respond different by adopting their highlights phenotypic heterogeneity plasticity epithelial cells.