Automated tracking of tumor-stroma morphology in microtissues identifies functional targets within the tumor microenvironment for therapeutic intervention

Abstract

Cancer-associated fibroblasts (CAFs) constitute an important part of the tumor microenvironment and promote invasion via paracrine functions and physical impact on the tumor. Although the importance of including CAFs into three-dimensional (3D) cell cultures has been acknowledged, computational support for quantitative live-cell measurements of complex cell cultures has been lacking. Here, we have developed a novel automated pipeline to model tumor-stroma interplay, track motility and quantify morphological changes of 3D co-cultures, in real-time live-cell settings. The platform consists of microtissues from prostate cancer cells, combined with CAFs in extracellular matrix that allows biochemical perturbation. Tracking of fibroblast dynamics revealed that CAFs guided the way for tumor cells to invade and increased the growth and invasiveness of tumor organoids. We utilized the platform to determine the efficacy of inhibitors in prostate cancer and the associated tumor microenvironment as a functional unit. Interestingly, certain inhibitors selectively disrupted tumor-CAF interactions, e.g. focal adhesion kinase (FAK) inhibitors specifically blocked tumor growth and invasion concurrently with fibroblast spreading and motility. This complex phenotype was not detected in other standard in vitro models. These results highlight the advantage of our approach, which recapitulates tumor histology and can significantly improve cancer target validation in vitro.

Proteomic alterations of fibroblasts induced by ovarian cancer cells reveal potential cancer targets

“…[32]. The tumor microenvironment should be considered as a target for cancer prevention and treatment, such as the blocking of interactions between cancer cells and cancerassociated fibroblasts [8,32].…”

“…It is now evident that cancer-associated fibroblasts secrete signaling factors to promote cancer development and modify the tumor microenvironment [6,7]. Cancer-associated fibroblasts have been shown to guide prostate cancer cells to invade and promote tumor organoids growth and invasion in a three-dimensional cell culture model [8]. Importantly, the protein alterations that exist within fibroblasts impact tumor progression.…”

The potential of organoids in urological cancer research

“…Moreover, the osteoblastic cells and the prostate cancer cells in the organoid system closely mimicked the architecture of prostate cancer metastasis in bone. Åkerfelt and colleagues 101 used a real-time, live-cell platform consisting of tissue from prostate cancer cells and cancer-associated fibroblasts in extracellular matrix to quantitatively monitor the dynamics of cancer-associated fibroblasts in 3D cell cultures. Cancer-associated fibroblasts were observed to initiate tumour cell invasion and promote tumour organoid growth and invasiveness.…”

“…As pure epithelial cultures, organoids lack many cellular components that are contained within an in vivo system (such as stromal, vascular endothelial, or immune cells). The biology and behaviour of cancer can be influenced by microenvironmental factors including the cell types present, which can explain observations such as differences in drug sensitivity for the same cancer cells grown in 2D or 3D cultures or monoculture or coculture with fibroblasts 101 . Increasing emphasis is being placed on the role of tumour–stromal inter actions 123 and several models based on the combination of 3D organoid systems and stromal cells have been developed 99,101 , which could aid in this research.…”

“…The biology and behaviour of cancer can be influenced by microenvironmental factors including the cell types present, which can explain observations such as differences in drug sensitivity for the same cancer cells grown in 2D or 3D cultures or monoculture or coculture with fibroblasts 101 . Increasing emphasis is being placed on the role of tumour–stromal inter actions 123 and several models based on the combination of 3D organoid systems and stromal cells have been developed 99,101 , which could aid in this research. Models that incorporate multiple cell types, such as fibroblasts, immune cells, and endothelial cells, into 3D culture systems need to be established to reflect the effects of extracellular matrix, epithelial–stromal communications, cell–matrix interactions, and cell–cell crosstalk.…”