Comparative Evaluation of Chemotherapy and CAR-T Cell Therapy in Genetically Engineered Self-Sustaining Primary Pancreatic Cancer Organoids.

BACKGROUND:Broader research and translational use of patient-derived organoids (PDOs) in pancreatic ductal adenocarcinoma (PDAC) remain constrained by complex and costly culture requirements. Here, we developed a conditional transgenic strategy to generate self-sustaining primary PDAC organoids and explored its feasibility for therapeutic evaluation. METHODS:We established a proof-of-concept PDAC organoid biobank (n = 10). Using a doxycycline-inducible lentiviral Tet-On system, we engineered organoids to express essential cytokines RSPO1 and WNT3A, enabling growth in supplement-depleted media. We then performed exploratory assessments of FOLFIRINOX-mimetic chemotherapy and EPHA2-targeted CAR-T cells in transgenic organoids, together with histopathological and clinicopathological co-analysis. RESULTS:Doxycycline-induced RSPO1/WNT3A expression rescued the impaired growth and passaging caused by exogenous cytokine depletion. In simplified media, transgenic organoids retained patient-specific histological and molecular features. Sensitivity to FOLFIRINOX-mimetic treatment and EPHA2 CAR-T cells varied among different patients, and the in vitro responses showed exploratory concordance with the corresponding clinical courses in a small subset. CONCLUSION:This conditional transgenic approach reduces culture complexity while preserving key phenotypic and functional characteristics of PDAC organoids, providing a feasible proof-of-concept platform for exploratory evaluation of chemotherapy and immunotherapy responses.