Speaker
Description
Introduction
Cholangiocarcinoma (CCA) is a rare but deadly disease that arises from epithelial cells in the liver (intrahepatic) or the biliary tract (extrahepatic). The absence of specific symptoms leads to late diagnosis, which is associated with a poor prognosis due to disease progression, metastasis, and the emergence of drug resistance. The heterogeneity of CCA makes the drug discovery process longer and more complex, partly due to the lack of reliable in vitro models for preclinical studies 1. CCAs are characterized by a dense, desmoplastic microenvironment with complex crosstalk between various stromal components and the extracellular matrix 2,3. Designing an in vitro model that mimics physiological conditions could improve the evaluation of drug efficacy.
Methods
To model the CCA tumor niche, gelatin methacryloyl (GelMA) hydrogels served as a 3D supporting matrix for co-culture of an immortalized CCA cell line (TFK-1) and human Mesenchymal Stem Cells (hMSCs). Matrix effects on cell behavior were investigated by assessing cell proliferation (viability assays), 3D structure formation (immunofluorescence), and gene expression (digital PCR). Furthermore, the influence of matrix composition on drug sensitivity was evaluated by assessing the effects of Gemcitabine and Curcumin 4 on cell viability.
Results
Consistent with the design of supportive matrices, high cell viability was observed, along with the formation of tumor spheroids within seven days. Furthermore, the matrix stiffness appeared to play a significant role, as softer hydrogels supported higher proliferation. Overall, RNA expression analyses revealed the maintenance of the epithelial phenotype in TFK-1 cells, demonstrating that methacryloyl residues and the photopolymerization process did not induce phenotypic modifications. On the other hand, the presence of hMSCs promoted the expression of genes associated with ECM remodeling (e.g., MMP9) and signaling pathways (e.g., VEGF). Furthermore, when the established 3D cultures were challenged with chemotherapeutic agents, we observed reduced sensitivity compared to 2D cultures, with the specific matrix influencing the response.
Discussion
Taken together, this study demonstrates the suitability of GelMA hydrogels for modeling the CCA niche, revealing significant influences of matrix stiffness on cell proliferation and drug sensitivity that are not observed in 2D cultures. These initial findings warrant further investigation into the specific matrix cues driving these differential responses and the potential for exploiting these interactions for improved therapeutic interventions in CCA.
References
1. Massa, A. et al. Evolution of the experimental models of cholangiocarcinoma. Cancers vol. 12 1–31 (2020).
2. Cadamuro, M. et al. The deleterious interplay between tumor epithelia and stroma in cholangiocarcinoma Biochim Biophys Acta Mol Basis Dis 1435–1443 (2018)
3. Affo, S. et al. Promotion of cholangiocarcinoma growth by diverse cancer-associated fibroblast subpopulations. Cancer Cell 39, 866-882.e11 (2021).
4. San, T. T. et al. Curcumin enhances chemotherapeutic effects and suppresses ANGPTL4 in anoikis-resistant cholangiocarcinoma cells. Heliyon 6, (2020).
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