Inhibiting TGFß-1 pathway reduces the aggressiveness of intrahepatic CCA HuCCT1 CD90 positive cells

Molecular mechanisms responsible for the poor prognosis in patients with intrahepatic cholangiocarcinoma (CCA) are still unknown, however, the stem cell marker cluster differentiation 90 (CD90), has been reported to be associated with a more aggressive cancer phenotype. In this scenario, TGFß1 signaling pathway likely has a role as master gene regulator. Aim of the study is to investigate the role of CD90 in iCCA aggressiveness. The molecular profile of HuCCT1/CD90+ and HuCCT1/CD90- cells was obtained through transcriptomic analysis (NGS). Bioinformatic data were confirmed in both cell lines by qRT-PCR and western blot. Cells were treated with Gemcitabine in monotherapy or in combination with Galunisertib, a selective TGFßRI inhibitor in 2D and 3D models. HuCCT1 CD90 positive cells are more proliferative, less migratory, and resistant to Gemcitabine treatment. Next, HuCCT1/CD90+ express lower levels of TGFß1 compared to /CD90- cell lines. Finally, HuCCT1/CD90+ are resistant to Gemcitabine, while the combination of both Gemcitabine and Galunisertib displays a synergistic effect on HuCCT1/CD90+ cell proliferation. These results underline that CD90 inducing Gemcitabine resistance can be overcome by adding a TGFß1 inhibitor such as Galunisertib, thereby moving further toward a precision medicine approach in patients with iCCA. Overall design: We used RNA sequencing technology to understand the molecular changes of CD90 increase in iCCA cell line