Integrated Single-Cell, Proteomic, and Tissue-Level Analysis Defines Cancer-Associated Fibroblast Programs in Pseudomyxoma Peritonei

CAF are abundantly present in fibrous septae containing the mucin and regions of inflammation. We established PMP-derived CAF cultures and characterized them through multiple omics and phenotypic assays.

Single-cell transcriptomic profiling revealed two distinct CAF populations: fibroblast-derived CAFs enriched for characteristic CAF functionalities and mesothelial-derived CAFs expressing mesothelial markers and pathways linked to metabolism and motility, consistent with mesothelial-to-mesenchymal transition. PMP CAFs exhibit baseline invasive and contractile properties and acquire an inflammatory CAF-like phenotype upon TNF-α/IL-1β stimulation, accompanied by altered proteomic and secretome profiles.