Transforming Growth Factors α and β Are Essential for Modeling Cholangiocarcinoma Desmoplasia and Progression in a Three-Dimensional Organotypic Culture Model.
To gain insight into the cellular and molecular interactions mediating the desmoplastic reaction and aggressive malignancy of mass-forming intrahepatic cholangiocarcinoma (ICC), we modeled ICC desmoplasia and progression in vitro. A unique three-dimensional (3D) organotypic culture model was established; within a dilute collagen-type I hydrogel, a novel clonal strain of rat cancer-associated myofibroblasts (TDFSM) was co-cultured with a pure rat cholangiocarcinoma cell strain (TDECC) derived from the same ICC type as TDFSM. This 3D organotypic culture model reproduced key features of desmoplastic reaction that closely mimicked those of the in situ tumor, as well as promoted cholangiocarcinoma cell growth and progression. Our results supported a resident liver mesenchymal cell origin of the TDFSMcells, which were not neoplastically transformed. Notably, 3D co-culturing of TDECCcells with TDFSMcells provoked the formation of a dense fibrocollagenous stroma in vitro that was associated with significant increases in both proliferative TDFSMmyofibroblastic cells and TDECCcholangiocarcinoma cells accumulating within the gel matrix. This dramatic desmoplastic ICC-like phenotype, which was not observed in the TDECCor TDFSMcontrols, was highly dependent on transforming growth factor (TGF)-β, but not promoted by TGF-α. However, TGF-α was determined to be a key factor for promoting cholangiocarcinoma cell anaplasia, hyperproliferation, and higher malignant grading in this 3D culture model of desmoplastic ICC.
Division of Cellular and Molecular Pathogenesis, Department of Pathology, Virginia Commonwealth University, Richmond, Virginia.
Miguel Á
MÁ
Manzanares
Division of Cellular and Molecular Pathogenesis, Department of Pathology, Virginia Commonwealth University, Richmond, Virginia.
Akihiro
A
Usui
Division of Cellular and Molecular Pathogenesis, Department of Pathology, Virginia Commonwealth University, Richmond, Virginia.
Deanna J
DJ
Campbell
Division of Cellular and Molecular Pathogenesis, Department of Pathology, Virginia Commonwealth University, Richmond, Virginia.
Catherine I
CI
Dumur
Division of Cellular and Molecular Pathogenesis, Department of Pathology, Virginia Commonwealth University, Richmond, Virginia.
Gabrielle T
GT
Maldonado
Division of Gastroenterology and Hepatology, University of Arkansas for Medical Sciences, Little Rock, Arkansas.
Michel
M
Fausther
Division of Gastroenterology and Hepatology, University of Arkansas for Medical Sciences, Little Rock, Arkansas.
Jonathan A
JA
Dranoff
Division of Cellular and Molecular Pathogenesis, Department of Pathology, Virginia Commonwealth University, Richmond, Virginia. Electronic address: alphonse.sirica@vcuhealth.org.
Alphonse E
AE
Sirica
D000818:Animals; D001650:Bile Duct Neoplasms / Q000209:etiology*; D001653:Bile Ducts, Intrahepatic / Q000378:metabolism*; D018281:Cholangiocarcinoma / Q000209:etiology*; D018920:Coculture Techniques; D004195:Disease Models, Animal; D018450:Disease Progression; D006801:Humans; D059785:Karyotype; D008297:Male; D058628:Myofibroblasts / Q000378:metabolism; D011916:Rats, Inbred F344; D016211:Transforming Growth Factor alpha / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D016211:Transforming Growth Factor alpha; D016212:Transforming Growth Factor beta