Cancer-Associated Fibroblasts in a 3D Engineered Tissue Model Induce Tumor-like Matrix Stiffening and EMT Transition

Abstract

A tumor microenvironment is characterized by its altered mechanical properties. However, most models remain unable to faithfully recreate the mechanical properties of a tumor. Engineered models based on the self-assembly method have the potential to better recapitulate the stroma architecture and composition. Here, we used the self-assembly method based on a bladder tissue model to engineer a tumor-like environment. The tissue-engineered tumor models were reconstituted from stroma-derived healthy primary fibroblasts (HFs) induced into cancer-associated fibroblast cells (iCAFs) along with an urothelium overlay. The iCAFs-derived extracellular matrix (ECM) composition was found to be stiffer, with increased ECM deposition and remodeling. The urothelial cells overlaid on the iCAFs-derived ECM were more contractile, as measured by quantitative polarization microscopy, and displayed increased YAP nuclear translocation. We further showed that the proliferation and expression of epithelial-to-mesenchymal transition (EMT) marker in the urothelial cells correlate with the increased stiffness of the iCAFs-derived ECM. Our data showed an increased expression of EMT markers within the urothelium on the iCAFs-derived ECM. Together, our results demonstrate that our tissue-engineered tumor model can achieve stiffness levels comparable to that of a bladder tumor, while triggering a tumor-like response from the urothelium.

Modulating extracellular matrix stiffness: a strategic approach to boost cancer immunotherapy

“…Signals originating from ECM stiffness initiate EMT, driving the transformation of cell morphology towards a mesenchymal phenotype and enhancing their invasive and migratory behaviors [ 45 ]. Specifically, ECM stiffness modulates cancer cell migration and invasion by promoting the expression of N-cadherin, while concurrently reducing the levels of E-cadherin [ 46 – 48 ]. Furthermore, enzymes responsible for ECM remodeling, which often exhibit increased expression during EMT, are prevalent in aggressive cancer phenotypes [ 49 ].…”

Stromal cells support the survival of human primary chronic lymphocytic leukemia (CLL) cells through Lyn-driven extracellular vesicles

“…Because it is also released in EVs, it might even link structural elements of the ECM beyond the cell membrane. CAF produce an extracellular matrix with a higher density as compared to normal fibroblasts and a high ECM density is an important driver of the tumor growth (35). Thus, mechanisms that contribute to ECM stiffness might also contribute to the interaction between tumor and bystander cells.…”

3D Tumor Models in Urology

“…When the tubular basement membrane of kidney tubules was stiffened, differentiation markers changed and pro-fibrotic growth factors increased [ 125 ] suggesting an impact on carcinogenic processes, too. Millet et al [ 126 ] highlighted the role of CAFs inducing matrix stiffening and EMT in a 3D self-engineered bladder model that displayed tumor-promoting effects of CAFs on normal urothelial cells as well.…”