Mimicking Tumor Cell Heterogeneity of Colorectal Cancer in a Patient-derived Organoid-Fibroblast Model

• Authors: ATANASOVA Velina S; CARDONA Crhistian de Jesus; HEJRET Václav; TIEFENBACHER Andreas; MAIR Theresia; TRAN Loan; PFNEISSL Janette; DRAGANIC Kristina; BINDER Carina; KABILJO Julijan; CLEMENT Janik; WOERAN Katharina; NEUDERT Barbara; WOHLHAUPTER Sabrina; HAASE Astrid; DOMAZET Sandra; HENGSTSCHLAEGER Markus; MITTERHAUSER Markus; MUELLAUER Leonhard; TICHÝ Boris; BERGMANN Michael; SCHWEIKERT Gabriele; HARTL Markus; DOLZNIG Helmut; EGGER Gerda
• Year of publication: 2023
• Type: Article in Periodical
• Magazine / Source: CELLULAR AND MOLECULAR GASTROENTEROLOGY AND HEPATOLOGY
• MU Faculty or unit: Central European Institute of Technology
• Web: https://www.sciencedirect.com/science/article/pii/S2352345X2300036X?via%3Dihub
• Doi: http://dx.doi.org/10.1016/j.jcmgh.2023.02.014
• Keywords: Colorectal Cancer; Fibroblasts; Organoids; targeted therapy
• Description: BACKGROUND & AIMS: Patient-derived organoid cancer models are generated from epithelial tumor cells and reflect tumor characteristics. However, they lack the complexity of the tumor microenvironment, which is a key driver of tumorigenesis and therapy response. Here, we developed a colorectal cancer organoid model that incorporates matched epithelial cells and stromal fibroblasts. METHODS: Primary fibroblasts and tumor cells were isolated from colorectal cancer specimens. Fibroblasts were character-ized for their proteome, secretome, and gene expression sig-natures. Fibroblast/organoid co-cultures were analyzed by immunohistochemistry and compared with their tissue of origin, as well as on gene expression levels compared with standard organoid models. Bioinformatics deconvolution was used to calculate cellular proportions of cell subsets in orga-noids based on single-cell RNA sequencing data. RESULTS: Normal primary fibroblasts, isolated from tumor adjacent tissue, and cancer associated fibroblasts retained their molecular characteristics in vitro, including higher motility of cancer associated compared with normal fibroblasts. Impor-tantly, both cancer-associated fibroblasts and normal fibro-blasts supported cancer cell proliferation in 3D co-cultures, without the addition of classical niche factors. Organoids grown together with fibroblasts displayed a larger cellular heteroge-neity of tumor cells compared with mono-cultures and closely resembled the in vivo tumor morphology. Additionally, we observed a mutual crosstalk between tumor cells and fibro-blasts in the co-cultures. This was manifested by considerably deregulated pathways such as cell-cell communication and extracellular matrix remodeling in the organoids. Thrombospondin-1 was identified as a critical factor for fibro-blast invasiveness. CONCLUSION: We developed a physiological tumor/stroma model, which will be vital as a personalized tumor model to study disease mechanisms and therapy response in colorectal cancer. (Cell Mol Gastroenterol Hepatol 2023;15:1391-1419; https://doi.org/10.1016/j.jcmgh.2023.02.014)