Scaffold-free and scaffold-based 3D models of intrahepatic cholangiocarcinoma for biomarker discovery and drug testing

Abstract

Intrahepatic cholangiocarcinoma (iCC) is highly resistant to standard chemotherapy, and despite numerous clinical trials, patient outcomes remain poor. This highlights the urgent need for physiologically relevant models to support target discovery and preclinical drug testing.

To address this, we have developed advanced 3D culture systems:

1. Self-organized branching iCC organoids (SoBOs)
2. Collagen-based 3D iCC cultures (CobaCs)

We also generated spheroids of intrahepatic biliary epithelial cell (HiCtrlS) as a non-malignant control. Together, these platforms provide a robust framework to investigate iCC drivers, including NDRG1, a stress-responsive protein implicated in multiple cancers but not yet extensively characterized in iCC.

Preliminary results show increased NDRG1 expression in CobaCs compared to parental 2D cultures. We are now investigating whether this upregulation also occurs in SoBOs and whether it reflects a response to 3D organization, including adaptation to hypoxia. Our goal is to elucidate NDRG1 regulation in these 3D systems and define its role in tissue remodeling and chemoresistance.

Impact:

Unlike conventional 2D cultures, our 3D models recapitulate tumor architecture and microenvironmental stress, providing a physiologically relevant platform. Integrating SoBOs and CobaCs with HiCtrlS allows discrimination of tumor-specific pathways from normal biliary biology. By uncovering key regulators of tumor progression, these models provide a foundation for developing more effective and personalized therapies for iCC patients.

Pipeline