Functional antagonism between STAT3 and SMAD4 regulates EMT

Oncogenic mutations in KRAS are among the most common in cancer. Classical models suggest that loss of epithelial characteristics and the acquisition of mesenchymal traits are associated with cancer aggressiveness and therapy resistance. We identify STAT3 as a genetic modifier of TGF-beta-induced epithelial to mesenchymal transition in mutant KRAS tumors. RNA sequencing was performed with murine cells expressing mutant KRAS either overexpressing hyperactive STAT3Y640, or CRISPR-mediated knockout of STAT3, SMAD4, or KRAS. Excel files of differential expression compared to control mutant RAS cells or fpkm files are provided., Total RNA was isolated using the PureLink RNA kit (ThermoFisher) and phenol-extracted. Whole exome RNA sequencing with bioinformatics was performed by Novogene Corp.. Excel data files include: 1) KRASG12D:p53null mouse embryo fibroblasts (MEF KP)(Ischenko et al. 2013 PNAS) and MEF KP expressing hyperactive STAT3 Y640F, 2) Murine KRASG12D:p53R172H PDAC (KPC)(Hingorani et al. 2005 Cancer Cell) and KPC KRAS knockout cells, 3) KPC and KPC STAT3 knockout cells, 4) KPC SMAD4 knockout cells., , # Functional antagonism between STAT3 and SMAD4 regulates EMT

### Contributors: Stephen DAmico, Varvara Kirillov, Oleski Petrenko, and Nancy Reich

**Description**: We evaluated STAT3 as a cancer dependency in KRAS-mediated tumorigenesis with established models using mouse embryonic fibroblasts and pancreatic ductal adenocarcinoma cells. The data highlight antagonistic epistasis between SMAD4 and STAT3, where SMAD4 expressing tumors are poorly differentiated in the absence of STAT3, while SMAD4 deficient tumors are well-differentiated in the presence of STAT3.

**Techniques**: Data include RNA Sequencing analysis of genetically modified tumorigenic cells lines. Total RNA was isolated using the PureLink RNA kit (ThermoFisher) and phenol-extracted. Whole exome RNA sequencing with bioinformatics was performed by Novogene Corp..

Excel data files are included:

1\) Gene ID, log fold change, and fpkm for RNA expression in KRASG12D:p53null mouse embryo fibroblasts (MEF KP)(Ischenko et al. 20...