title: 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.
authors.affiliation: Turku Centre for Biotechnology, University of Turku, Turku, FI-20520, Finland.|VTT Technical Research Centre of Finland, Turku, FI-20521, Finland.
authors.forename: Malin
authors.initials: M
authors.lastname: Åkerfelt
authors.affiliation: Centre for Machine Vision Research, University of Oulu, Oulu, FI-90014, Finland.
authors.forename: Neslihan
authors.initials: N
authors.lastname: Bayramoglu
authors.affiliation: Department of Mathematics and Statistics, University of Turku, Turku, FI-20014, Finland.|University Grenoble Alpes, iRTSV-BGE, Grenoble, F-38000, France.|CEA, iRTSV-BGE, Grenoble, F-38000, France.|INSERM, BGE, Grenoble, F-38000, France.
authors.forename: Sean
authors.initials: S
authors.lastname: Robinson
authors.affiliation: Turku Centre for Biotechnology, University of Turku, Turku, FI-20520, Finland.|VTT Technical Research Centre of Finland, Turku, FI-20521, Finland.|Institute of Biomedicine, University of Turku, Turku, FI-20520, Finland.
authors.forename: Mervi
authors.initials: M
authors.lastname: Toriseva
authors.affiliation: Institute of Biomedicine, University of Turku, Turku, FI-20520, Finland.
authors.forename: Hannu-Pekka
authors.initials: HP
authors.lastname: Schukov
authors.affiliation: VTT Technical Research Centre of Finland, Turku, FI-20521, Finland.
authors.forename: Ville
authors.initials: V
authors.lastname: Härmä
authors.affiliation: VTT Technical Research Centre of Finland, Turku, FI-20521, Finland.
authors.forename: Johannes
authors.initials: J
authors.lastname: Virtanen
authors.affiliation: Biocenter Oulu and Department of Pathology, University of Oulu and Oulu University Hospital, Oulu, FI-90220, Finland.
authors.forename: Raija
authors.initials: R
authors.lastname: Sormunen
authors.affiliation: Oulu Center for Cell-Matrix Research, Biocenter Oulu and Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, FI-90014, Finland.
authors.forename: Mika
authors.initials: M
authors.lastname: Kaakinen
authors.affiliation: Centre for Machine Vision Research, University of Oulu, Oulu, FI-90014, Finland.
authors.forename: Juho
authors.initials: J
authors.lastname: Kannala
authors.affiliation: Oulu Center for Cell-Matrix Research, Biocenter Oulu and Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, FI-90014, Finland.
authors.forename: Lauri
authors.initials: L
authors.lastname: Eklund
authors.affiliation: Centre for Machine Vision Research, University of Oulu, Oulu, FI-90014, Finland.
authors.forename: Janne
authors.initials: J
authors.lastname: Heikkilä
authors.affiliation: Turku Centre for Biotechnology, University of Turku, Turku, FI-20520, Finland.|VTT Technical Research Centre of Finland, Turku, FI-20521, Finland.
authors.forename: Matthias
authors.initials: M
authors.lastname: Nees
mesh_terms: D000465:Algorithms; D002450:Cell Communication / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D058948:Cell Tracking / Q000379:methods*; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D008297:Male; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D008954:Models, Biological; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D059008:Time-Lapse Imaging / Q000379:methods*; D059016:Tumor Microenvironment
chemical_list: D047428:Protein Kinase Inhibitors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D003094:Collagen; D051416:Focal Adhesion Protein-Tyrosine Kinases
keywords: 3D co-culture; cancer associated fibroblast (CAF); focal adhesion kinase (FAK); invasion; phenotypic screening