TY - JOUR
T1 - Mimicking tumor cell heterogeneity of colorectal cancer in a patient-derived organoid-fibroblast model
AU - Atanasova, Velina S.
AU - de Jesus Cardona, Crhistian
AU - Hejret, Václav
AU - Tiefenbacher, Andreas
AU - Mair, Theresia
AU - Tran, Loan
AU - Pfneissl, Janette
AU - Draganić, Kristina
AU - Binder, Carina
AU - Kabiljo, Julijan
AU - Clement, Janik
AU - Woeran, Katharina
AU - Neudert, Barbara
AU - Wohlhaupter, Sabrina
AU - Haase, Astrid
AU - Domazet, Sandra
AU - Hengstschläger, Markus
AU - Mitterhauser, Markus
AU - Müllauer, Leonhard
AU - Tichý, Boris
AU - Bergmann, Michael
AU - Schweikert, Gabriele
AU - Hartl, Markus
AU - Dolznig, Helmut
AU - Egger, Gerda
N1 - Funding Information:
Austrian Science Foundation (FWF), doc.funds grant DOC59 (GE, TM) Austrian Science Foundation (FWF) SFB F83 (GE, JF, KD) City of Vienna Fund for Innovative Interdisciplinary Cancer Research, 21118 (GE) FFG-Industrienahe Dissertationen 879481(GE, MB, VSA, JK) Austrian Academy of Sciences, Doc Fellowship 25276 (LT) CCC research grant of the MedUni Vienna (HD) European Union, SECRET‐ITN 859962 (HD) Niederösterreichische Forschungs‐ und Bildungsges.m.bH.; NFB, LSC18‐017 (HD) BMBF, German Network for Bioinformatics Infrastructure (de.NBI) 031A537B, 031A533A, 031A538A, 031A533B, 031A535A, 031A537C, 031A534A, 031A532B (CJC, GS).
Copyright:
© 2023 The Authors. Published by Elsevier Inc.
PY - 2023
Y1 - 2023
N2 - 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 characterized for their proteome, secretome, and gene expression signatures. 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 organoids 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. Importantly, both cancer-associated fibroblasts and normal fibroblasts supported cancer cell proliferation in 3D co-cultures, without the addition of classical niche factors. Organoids grown together with fibroblasts displayed a larger cellular heterogeneity 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 fibroblasts 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 fibroblast 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.
AB - 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 characterized for their proteome, secretome, and gene expression signatures. 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 organoids 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. Importantly, both cancer-associated fibroblasts and normal fibroblasts supported cancer cell proliferation in 3D co-cultures, without the addition of classical niche factors. Organoids grown together with fibroblasts displayed a larger cellular heterogeneity 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 fibroblasts 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 fibroblast 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.
KW - colorectal cancer
KW - organoids
KW - co-cultures
KW - fibroblasts
KW - Cancer
KW - Organoids
KW - Colorectal Cancer
KW - Fibroblasts
KW - Co-cultures
UR - http://www.scopus.com/inward/record.url?scp=85152918456&partnerID=8YFLogxK
U2 - 10.1016/j.jcmgh.2023.02.014
DO - 10.1016/j.jcmgh.2023.02.014
M3 - Article
C2 - 36868311
SN - 2352-345X
VL - 15
SP - 1391
EP - 1419
JO - Cellular and Molecular Gastroenterology and Hepatology
JF - Cellular and Molecular Gastroenterology and Hepatology
IS - 6
ER -
