mRNA gene expression of colon fibroblasts isolated from mouse colon

The EMT-transcription factor ZEB1 is heterogeneously expressed in tumor cells and in cancer-associated fibroblasts (CAFs) in colorectal cancer (CRC). While ZEB1 in tumor cells regulates metastasis and therapy resistance, its role in CAFs is largely unknown. Combining fibroblast-specific Zeb1 deletion with immunocompetent mouse models of CRC, we observe that inflammation-driven tumorigenesis is accelerated, whereas invasion and metastasis in sporadic cancers is reduced upon fibroblast-specific loss of Zeb1. Single-cell transcriptomics, histological characterization and in vitro modelling reveal a crucial role in CAF polarization, promoting myofibroblastic features by restricting inflammatory activation. Zeb1 deficiency impairs collagen deposition and CAF barrier function but increasesd NF?B-mediated cytokine production, jointly promoting lymphocyte recruitment and immune checkpoint activation. Overall design: Established fibroblast cultures from Zeb1 flox/flox (Fib-Ctrl) and Zeb1 del/del (Fib-?Zeb1) were cultured regularly and RNA was harvested by Qiagen RNeasy