Prrx1 is a Master Transcription Factor of myofibroblastic Cancer-Associated Fibroblasts [Human_CAF_ChIP_seq]

Although cancer associated fibroblasts (CAF) play a critical role in cancer progression, their identities are still unknown. Because cells' own identity is determined by core regulatory circuit (CRC) comprising master transcription factors (TFs), it is critical to find CAF's master TF. Therefore, we attempt to find master TF of CAF through the extensive screening of single cell RNA-seq database of cancer tissues, and discovered Prrx1, master regulator of embryonic mesenchymal cells, as candidate for master TF of CAF. Prrx1 was abundantly expressed in CAFs with significant correlation with unfavorable clinical outcomes. Furthermore, Prrx1 in fibroblasts dramatically enhanced tumor progression and metastasis in fibroblast-specific Prrx1 inducible mice. On the other hand, tumor formation was severely inhibited in conditional Prrx1 knock-down mice. Finally, we confirmed that Prrx1 is a master TF of CAF and normal fibroblast executing wound healing program using ChIP-SEQ, CRC modeling, and in vivo validation. Furthermore we also revealed that targeting Prrx1 in fibroblast alone induced complete remission of chemotherapy-resistant cancer in preclinical mouse experiments. Overall design: First, three murine fibroblasts such as mouse embryonic fibroblast, wound healing fibroblast, and MMTV CAF are obtained from C57BL/6, FVB strains. The ChIP-seq were performed in these fibroblasts using H3K27ac, H3K4me3, and Prrx1 antibody and the RNA-seq also were performed. Second, single cells of murine wound tissue obtained from BALB/c were performed using 10X platform. Third, nine ex vivo cultured human CAFs isolated from advanced colorectal cancer tissue were established. we performed H3K27ac ChIP-seq and RNA-seq from these human CAFs.