Dataset Information

Bulk RNA-seq of WNT-responsive and WNT-secreting cancer cell populations from orthotopic PDAC tumors

ABSTRACT: Pancreatic ductal adenocarcinoma (PDAC) exhibits pronounced intratumoral heterogeneity driven in part by WNT signaling dynamics. To characterize transcriptional differences among distinct WNT signaling states in vivo, we performed bulk RNA sequencing of fluorescence-activated cell sorting (FACS)-isolated cancer cell populations from orthotopic PDAC tumors. WNT-responsive (WNT-R), WNT-secreting (WNT-S), double-positive (WNT-R/S), and double-negative (DN) cancer cells were isolated from KPC 7TCF-eGFP/Porcn-DMA reporter tumors. RNA libraries were prepared using the SMARTerSeq2 protocol and sequenced from four biological replicates per group. Differential gene expression analysis revealed enrichment of canonical WNT/β-catenin target genes in WNT-R cells, elevated Porcn expression in WNT-S cells, and intermediate transcriptional profiles in WNT-R/S cells, validating the fidelity of the reporter system in vivo. This dataset provides a resource for dissecting WNT-dependent cancer cell states and signaling interactions in PDAC.

ORGANISM(S): Mus musculus

PROVIDER: GSE316194 | GEO | 2026/05/20

REPOSITORIES: GEO

ACCESS DATA

Similar Datasets

Project description:Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease characterized by late diagnosis and treatment resistance. Recurrent genetic alterations in defined genes in association with perturbations of developmental cell signaling pathways have been associated with PDAC development and progression. Here, we show that GATA6 contributes to pancreatic carcinogenesis during the temporal progression of pancreatic intraepithelial neoplasia by virtue of Wnt pathway activation. GATA6 is recurrently amplified by both quantitative-PCR and fluorescent in-situ hybridization in human pancreatic intraepithelial neoplasia and in PDAC tissues, and GATA6 copy number is significantly correlated with overall patient survival. Forced overexpression of GATA6 in cancer cell lines enhanced cell proliferation and colony formation in soft agar in vitro and growth in vivo, as well as increased Wnt signaling. By contrast siRNA mediated knockdown of GATA6 led to corresponding decreases in these same parameters. The effects of GATA6 were found to be due to its ability to bind DNA, as forced overexpression of a DNA-binding mutant of GATA6 had no effects on cell growth in vitro or in vivo, nor did they affect Wnt signaling levels in these same cells. A microarray analysis revealed the Wnt antagonist Dickopf-1 (DKK1) as a dysregulated gene in association with GATA6 knockdown, and direct binding of GATA6 to the DKK1 promoter was confirmed by chromatin immunoprecipitation and electrophoretic mobility shift assays. Transient transfection of GATA6, but not mutant GATA6, into cancer cell lines led to decreased DKK1 mRNA expression and secretion of DKK1 protein into culture media. Forced overexpression of DKK1 antagonized the effects of GATA6 on Wnt signaling in pancreatic cancer cells. These findings illustrate that one mechanism by which GATA6 promotes pancreatic intraepithelial neoplasia is by virtue of its activation of canonical Wnt signaling via regulation of DKK1. AsPC1 and A13A cells were stably transfected with a lentivirus expressing mock shRNA or shRNA to GATA6. Each control/shRNA pair (total 4 samples) was analyzed by two-color microarray and the genes commonly dysregulated in both cell lines identified.

Dataset Information

Bulk RNA-seq of WNT-responsive and WNT-secreting cancer cell populations from orthotopic PDAC tumors

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets