Gene expression of Cancer-associated fibroblasts from AOM/DSS autchthonous murine tumors on the single cell level (Sort-Seq)

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 NFκB-mediated cytokine production, jointly promoting lymphocyte recruitment and immune checkpoint activation. We used single cell RNA sequencing (scRNA-seq) via SORT-Seq to analyze the diversity of murine AOM/DSS CAFs enriched by FACS (following colon tumor digestion) by gating for absence of Epcam, CD45 and CD31 immunoreactivity.