Cholangiocarcinoma (CCA) is a markedly desmoplastic tumor, with extrahepatic CCA (eCCA) being the most prevalent subtype. The nonspecific clinical manifestation and early metastatic potential result in a persistently poor overall prognosis. Cancer-associated fibroblasts (CAFs) are one of the major components of the tumor microenvironment implicated in tumor progression and treatment resistance. To faithfully recapitulate the eCCA microenvironment, we established a comprehensive patient-derived preclinical platform comprising CAFs, eCCA primary cells, organoids, and patient-derived xenograft (PDX) models. We found that CAFs facilitated eCCA cell growth, migration, and invasion in vivo and in vitro. Cytokine profiling revealed that HGF is a key paracrine factor produced by CAFs. CAF-derived HGF activates c-MET signaling to drive gemcitabine resistance in eCCA. Therapeutically, pharmacologic inhibition of c-MET partially resensitized eCCA cells to gemcitabine in preclinical models and suppressed tumor progression. In conclusion, our data provides subtype-specific translational evidence that therapeutically targeting the CAF-HGF-c-MET pathway represents a rational strategy to enhance gemcitabine efficacy and improve treatment outcomes in eCCA.

Extrahepatic cholangiocarcinoma  /  Primary cells  /  Cancer-associated fibroblasts  /  Organoids  /  Patient-derived xenograft  /  HGF  /  c-MET  /  Gemcitabine resistance