Mp65-09 Bladder Cancer Organoids: A Reliable Tool for Validation of in Vitro New Therapeutic Approaches

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III (MP65)1 May 2024MP65-09 BLADDER CANCER ORGANOIDS: A RELIABLE TOOL FOR VALIDATION OF IN VITRO NEW THERAPEUTIC APPROACHES Emilie Decaup, Claire Béraud, Anne Sophie Bajeot, Xavier Gamé, Nicolas Monjotin, Pascal Rischmann, Philippe Lluel, and Mathieu Roumiguié Emilie DecaupEmilie Decaup , Claire BéraudClaire Béraud , Anne Sophie BajeotAnne Sophie Bajeot , Xavier GaméXavier Gamé , Nicolas MonjotinNicolas Monjotin , Pascal RischmannPascal Rischmann , Philippe LluelPhilippe Lluel , and Mathieu RoumiguiéMathieu Roumiguié View All Author Informationhttps://doi.org/10.1097/01.JU.0001008756.24343.22.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: In patients with bladder cancer (BC) immune therapies have recently demonstrated an improvement for overall survival rates compared to chemotherapy alone. However, it remains an unmet need to develop novel therapeutic strategies. In this context, organoids have emerged as a promising animal-free tool revealing patients' heterogeneity and allowing large preclinical therapeutic screening. Urosphere developed a BC organoids biobank currently containing 8 models with phenotypic, pharmacological and molecular characterization. The aim of this study was to demonstrate the relevance of our organoids models to validate therapeutic approaches such as targeted or CAR-T cell therapies using EGFR as target. METHODS: Urothelial progenitors were isolated from tumour samples and seeded in Matrigel®. Omic data allowed us to select models with high transcriptomic EGFR expressions. To evaluate targeted-therapy, organoids were treated with erlotinib (0.1 to 10 µM) for 5 days. Cell viability was measured with CellTiter-Glo3D® assay. To validate CAR-T cell approach, organoids were co-cultured with anti-EGFR or control CAR-T cells at 4 effector:target ratios. Cell viability was measured with CellTiter-Glo3D® and Caspase-Glo® 3/7 3D assays. Pro-inflammatory and Granzyme B release were analysed by multiplex assay. RESULTS: Organoids from 2 models with high transcriptomic EGFR expression were selected and protein expression was confirmed by immunofluorescence. For targeted therapy, erlotinib presented significant efficacy in both models. In the model with high EGFR expression, erlotinib induced cell death up to 80% with an IC50 of 2.5 µM whereas in the lower-expressing model the IC50 was 21.4 µM. Concerning CAR-T cell therapy, after 48 h of co-culture with anti-EGFR CAR-T cells, a decrease of cell viability associated with an increase of apoptosis was observed. After 72 h of co-culture, an increase in the secretion of pro-inflammatory cytokines (IFN-g and TNF-a) and Granzyme B by anti-EGFR CAR-T cells was also observed. In co-cultures with organoids presenting high EGFR expression, CAR-T cells activation appeared higher with 14 fold release of IFN-g and TNF-a by anti-EGFR CAR-T cells compared to those co-cultured with organoids lower expressing EGFR. CONCLUSIONS: In an EGFR targeting approach, the use of organoids to evaluate both targeted- and CAR-T cell therapies was validated with concordant results. This robust translational model is fully transposable to other therapeutic approaches, whether in a traditional culture model or in a co-culture system. Source of Funding: Urosphere © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1080 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Emilie Decaup More articles by this author Claire Béraud More articles by this author Anne Sophie Bajeot More articles by this author Xavier Gamé More articles by this author Nicolas Monjotin More articles by this author Pascal Rischmann More articles by this author Philippe Lluel More articles by this author Mathieu Roumiguié More articles by this author Expand All Advertisement PDF downloadLoading ...