Integrated multi-omics and experimental evidence implicate DVL3-associated Wnt/β-catenin signaling in PFHxS-related colorectal cancer progression

Perfluorohexanesulfonic acid (PFHxS) is a persistent per- and polyfluoroalkyl substance (PFAS) widely detected in human biospecimens, yet its relationship with colorectal cancer (CRC) progression remains poorly defined. We integrated network toxicology, bulk transcriptomics, machine-learning-based prognostic modeling, single-cell and spatial transcriptomics, and functional experiments to characterize PFHxS-associated molecular features in CRC. Intersecting PFHxS-related targets with CRC-associated genes identified 707 shared targets, of which 203 core nodes were retained after protein–protein interaction and topological filtering. A 14-gene gradient boosting machine signature stratified overall survival in TCGA and two external cohorts and remained independently prognostic after adjustment for clinical covariates. Single-cell analysis and SHAP-based model attribution prioritized DVL3 for experimental investigation. Chronic PFHxS exposure increased DVL3 abundance and malignant phenotypes in CRC models. DVL3 knockdown attenuated PFHxS-associated clonogenic and invasive phenotypes and reduced Wnt/β-catenin transcriptional activity and downstream target-gene expression, with key findings reproduced in CT26 cells. Continuous in vivo PFHxS exposure increased xenograft tumor burden and relative Ki-67-positive cell counts, with stronger DVL3 staining in representative tumor sections. Collectively, these findings link PFHxS exposure to aggressive CRC phenotypes and support DVL3 as a functional contributor to Wnt/β-catenin transcriptional responses.