A patient-specific in vitro model for the study of metastatic bile duct cancer

A patient-specific in vitro model for the study of metastatic bile duct cancer: Combining decellularization of metastatic locations with patient-derived cholangiocarcinoma organoids

van Beek, Marije (TU Delft Mechanical, Maritime and Materials Engineering)

Fratila-Apachitei, E.L. (mentor)
Verstegen, M.M.A. (mentor)
van Tienderen, G.S. (graduation committee)

Delft University of Technology

Mechanical Engineering | BioMechanical Design

2023-02-28

Cholangiocarcinoma (CCA) is a rare but aggressive type of primary liver cancer with a dismal prognosis. CCA commonly metastasizes to the lungs and lymph nodes, significantly reducing overall survival. However, a mechanistic understanding of how CCA invades these metastatic sites remains lacking. This is partly due to the failure of current models to recapitulate the complexity of tissue-specific environments for metastatic CCA. Patient-derived tumor organoids are promising models for studying cancer in vitro, including CCA. However, organoids are frequently cultured in basement membrane extract (BME), which does not represent the native microenvironment. Decellularized scaffolds can serve as a culture environment for investigating tumor-extracellular matrix (ECM) interactions. Nonetheless, decellularized scaffolds are often utilized in conjunction with cancer cell lines, lacking critical characteristics of a growing tumor in vivo. Therefore, this study aims to decellularize and characterize human-derived lung (n=3) and lymph nodes (n=17), and recellularize these with patient-derived CCA organoids (CCAOs) (n=3) to establish an in vitro model for studying the interaction between epithelial tumor cells and the microenvironment of the metastatic site. Decellularization resulted in acellular scaffolds with preserved ECM components. Local ECM heterogeneity was shown by the macro- and micro-scale mechanical properties as determined by rheology and micro-indentation. The CCAOs showed adherence and growth when combining them with the decellularized lung ECM (dLECM). The metabolic activity of CCAOs was diminished when cultured in dLECM compared to those cultured in BME, and was dictated both by the organoid line and the dLECM donor. Distinct CCAO gene expression profiles were observed between the two culture environments (BME and dLECM). This elucidates the effect of the culture environment on the behavior of CCAOs in an in vitro model. In conclusion, the convergence of CCAOs with their organ-specific location of metastasis obtained by decellularization, provides a valuable tool for integrating the ECM of the metastatic location in an in vitro model for the mechanistic study of cancer metastasis.

decellularization
cholangiocarcinoma organoids
extracellular matrix
metastasis
recellularization

http://resolver.tudelft.nl/uuid:49ed2390-6d7d-4aee-88d6-8b6b98ecb276

2023-03-28

51.910144, 4.4660355

Student theses

master thesis

© 2023 Marije van Beek