Sebastian Schuth1, Solange Le Blanc1,2,3,4, Teresa G Krieger5, Julia Jabs6,7, Christian Conrad8,9, Oliver Strobel10,11,12, Miriam Schenk1, Nathalia A Giese1, Markus W Büchler1, Roland Eils5,6. 1. European Pancreas Center, Department of General Surgery, University Hospital Heidelberg, Heidelberg, Germany. 2. Division of Molecular Oncology of Gastrointestinal Tumors, German Cancer Research Center (DKFZ), Heidelberg, Germany. 3. NCT partner site Heidelberg, a clinical-translational cancer research partnership between University Hospital Heidelberg and DKFZ, Heidelberg, Germany. 4. Division of Visceral Surgery, Department of General Surgery, Medical University of Vienna, Vienna, Austria. 5. Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Digital Health Center, Berlin, Germany. 6. Division of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany. 7. Present Address: Merck Healthcare KGaA, Global Research, Darmstadt, Germany. 8. Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Digital Health Center, Berlin, Germany. christian.conrad@charite.de. 9. Division of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany. christian.conrad@charite.de. 10. European Pancreas Center, Department of General Surgery, University Hospital Heidelberg, Heidelberg, Germany. oliver.strobel@meduniwien.ac.at. 11. NCT partner site Heidelberg, a clinical-translational cancer research partnership between University Hospital Heidelberg and DKFZ, Heidelberg, Germany. oliver.strobel@meduniwien.ac.at. 12. Division of Visceral Surgery, Department of General Surgery, Medical University of Vienna, Vienna, Austria. oliver.strobel@meduniwien.ac.at.
Abstract
BACKGROUND: Cancer-associated fibroblasts (CAFs) are considered to play a fundamental role in pancreatic ductal adenocarcinoma (PDAC) progression and chemoresistance. Patient-derived organoids have demonstrated great potential as tumor avatars for drug response prediction in PDAC, yet they disregard the influence of stromal components on chemosensitivity. METHODS: We established direct three-dimensional (3D) co-cultures of primary PDAC organoids and patient-matched CAFs to investigate the effect of the fibroblastic compartment on sensitivity to gemcitabine, 5-fluorouracil and paclitaxel treatments using an image-based drug assay. Single-cell RNA sequencing was performed for three organoid/CAF pairs in mono- and co-culture to uncover transcriptional changes induced by tumor-stroma interaction. RESULTS: Upon co-culture with CAFs, we observed increased proliferation and reduced chemotherapy-induced cell death of PDAC organoids. Single-cell RNA sequencing data evidenced induction of a pro-inflammatory phenotype in CAFs in co-cultures. Organoids showed increased expression of genes associated with epithelial-to-mesenchymal transition (EMT) in co-cultures and several potential receptor-ligand interactions related to EMT were identified, supporting a key role of CAF-driven induction of EMT in PDAC chemoresistance. CONCLUSIONS: Our results demonstrate the potential of personalized PDAC co-cultures models not only for drug response profiling but also for unraveling the molecular mechanisms involved in the chemoresistance-supporting role of the tumor stroma.
BACKGROUND: Cancer-associated fibroblasts (CAFs) are considered to play a fundamental role in pancreatic ductal adenocarcinoma (PDAC) progression and chemoresistance. Patient-derived organoids have demonstrated great potential as tumor avatars for drug response prediction in PDAC, yet they disregard the influence of stromal components on chemosensitivity. METHODS: We established direct three-dimensional (3D) co-cultures of primary PDAC organoids and patient-matched CAFs to investigate the effect of the fibroblastic compartment on sensitivity to gemcitabine, 5-fluorouracil and paclitaxel treatments using an image-based drug assay. Single-cell RNA sequencing was performed for three organoid/CAF pairs in mono- and co-culture to uncover transcriptional changes induced by tumor-stroma interaction. RESULTS: Upon co-culture with CAFs, we observed increased proliferation and reduced chemotherapy-induced cell death of PDAC organoids. Single-cell RNA sequencing data evidenced induction of a pro-inflammatory phenotype in CAFs in co-cultures. Organoids showed increased expression of genes associated with epithelial-to-mesenchymal transition (EMT) in co-cultures and several potential receptor-ligand interactions related to EMT were identified, supporting a key role of CAF-driven induction of EMT in PDAC chemoresistance. CONCLUSIONS: Our results demonstrate the potential of personalized PDAC co-cultures models not only for drug response profiling but also for unraveling the molecular mechanisms involved in the chemoresistance-supporting role of the tumor stroma.
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