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// Malin Akerfelt 1, 2 , Neslihan Bayramoglu 3 , Sean Robinson 4, 5, 6, 7 , Mervi Toriseva 1, 2, 8 , Hannu-Pekka Schukov 8 , Ville Harma 2 , Johannes Virtanen 2 , Raija Sormunen 9 , Mika Kaakinen 10 , Juho Kannala 3 , Lauri Eklund 10 , Janne Heikkila 3 , Matthias Nees 1, 2 1 Turku Centre for Biotechnology, University of Turku, Turku, FI-20520, Finland 2 VTT Technical Research Centre of Finland, Turku, FI-20521, Finland 3 Centre for Machine Vision Research, University of Oulu, Oulu, FI-90014, Finland 4 Department of Mathematics and Statistics, University of Turku, Turku, FI-20014, Finland 5 University Grenoble Alpes, iRTSV-BGE, Grenoble, F-38000, France 6 CEA, iRTSV-BGE, Grenoble, F-38000, France 7 INSERM, BGE, Grenoble, F-38000, France 8 Institute of Biomedicine, University of Turku, Turku, FI-20520, Finland 9 Biocenter Oulu and Department of Pathology, University of Oulu and Oulu University Hospital, Oulu, FI-90220, Finland 10 Oulu Center for Cell-Matrix Research, Biocenter Oulu and Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, FI-90014, Finland Correspondence to: Matthias Nees, e-mail: matthias.nees@btk.fi Keywords: 3D co-culture, cancer associated fibroblast (CAF), phenotypic screening, invasion, focal adhesion kinase (FAK) Received: March 21, 2015 Accepted: August 24, 2015 Published: September 03, 2015 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 .

Paula Vainio 1 , Laura Lehtinen 1 , Tuomas Mirtti 2,3,4 , Mika Hilvo 5 , Tuulikki Seppanen-Laakso 5 , Johannes Virtanen 1 , Anna Sankila 4 , Stig Nordling 4 , Johan Lundin 3 , Antti Rannikko 6 , Matej Oresic 5 , Olli Kallioniemi 1,3 and Kristiina Iljin 1 1 Medical Biotechnology, VTT Technical Research Centre of Finland, and Turku Centre for Biotechnology, University of Turku, Finland 2 Haartman Institute, Department of Pathology, University of Helsinki, Finland 3 Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Finland 4 HUSLAB, Department of Pathology, Helsinki University Central Hospital, Finland 5 Bio and Process Technology, VTT Technical Research Centre of Finland, Espoo, Finland 6 Department of Urology, Helsinki University Central Hospital, Finland Received: December 20, 2011; Accepted: December 22, 2011; Published: December 22, 2011; Keywords: Prostate cancer, PLA2G7, drug target, biomarker, statins Correspondence: Kristiina Iljin, email: // // Abstract Prostate cancer is the second leading cause of cancer mortality in men in developed countries. Due to the heterogeneous nature of the disease, design of novel personalized treatments is required to achieve efficient therapeutic responses. We have recently identified phospholipase 2 group VII ( PLA2G7 ) as a potential drug target especially in ERG oncogene positive prostate cancers. Here, the expression profile of PLA2G7 was studied in 1137 prostate cancer and 409 adjacent non-malignant prostate tissues using immunohistochemistry to validate its biomarker potential and putative association with disease progression. In order to reveal the molecular alterations induced by PLA2G7 impairment, lipidomic and gene expression profiling was performed in response to PLA2G7 silencing in cultured prostate cancer cells. Moreover, the antineoplastic effect of statins combined with PLA2G7 impairment was studied in prostate cancer cells to evaluate the potential of repositioning of in vivo compatible drugs developed for other indications towards anti-cancer purposes. The results indicated that PLA2G7 is a cancer-selective biomarker in 50 % of prostate cancers and associates with aggressive disease. The alterations induced by PLA2G7 silencing highlighted the potential of PLA2G7 inhibition as an anti-proliferative, pro-apoptotic and anti-migratorial therapeutic approach in prostate cancer. Moreover, the anti-proliferative effect of PLA2G7 silencing was potentiated by lipid-lowering statins in prostate cancer cells. Taken together, our results support the potential of PLA2G7 as a biomarker and a drug target in prostate cancer and present a rationale for combining PLA2G7 inhibition with the use of statins in prostate cancer management.