Patient-derived lung cancer organoids as 3D-testing platform for precision CAR T-cell therapy
EHLEN L. 1,2, FARRERA-SAL M. 1, SZYSKA M. 1, ARNDT J. 1,2, SCHALLENBERG S. 3, THIEDE K. 1, SCHOLZ C. 1, GOLUSDA L. 1, MAI M. 1, SCHULENBERG S. 1, PETER L. 1, PICHT S. 1, LÖWA A. 4, VOLLBRECHT C. 3, JOOSTEN M. 3, HOCKE A. 4, SPIES C. 2, SCHMUECK-HENNERESSE M. 1
1 Berlin Institute of Health (BIH) at Charité - Universitätsmedizin Berlin, BIH Center for Regenerative Therapies (BCRT) , Berlin, Germany; 2 Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Berlin Institute of Health, Department of Anesthesiology and Intensive Care Medicine, Berlin, Germany; 3 Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Berlin Institute of Health, Institute of Pathology, Berlin, Germany; 4 Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Berlin Institute of Health, Department of Infectious Diseases and Respiratory Medicine, Berlin, Germany
Objectives: Lung cancer is the leading cause of cancer-related death globally. Despite the success of CAR T-cells in hematological diseases, their effectiveness in solid tumors, including lung cancer, is limited. To address individual resistance mechanisms, models representing tumor heterogeneity are necessary for personalized testing and improvement of immunotherapy in lung cancer patients. To address this, we have developed patient-derived lung cancer organoids (PDLCOs) from resected tumors, and CAR T-cells that can be adapted to individual PDLCO features.
Methods: Lung tumor samples were used to establish PDLCOs through dissociation, embedding in extracellular matrix, and growth factor supplementation. Comparative analysis of PDLCOs and parental tumors employed Histology, DNA sequencing, and single-cell RNA sequencing. CAR constructs were designed using a modular system with a single-chain library, non-viral CRISPR-based gene insertion was employed to generate CAR T-cells, and their efficacy in killing PDLCOs was evaluated using live cell imaging.
Results: We developed PDLCOs from resected tumors and matched healthy organoids for comparison and confirmed the fidelity of PDLCOs to parental tumors. Heterogeneous expression of lung tumor-specific tumor-associated antigens (TAAs) was observed in PDCLOs and parental tumors. To identify suitable TAAs for CAR T-cell therapy, a lung tumor specific TAA screening panel was created. CARs targeting these TAAs were engineered. Live cell imaging demonstrated specific killing of PDLCOs by CAR T-cells.
Conclusions: Our study established patient-derived organoids as avatars of parental tumors and utilized a modular CAR system for testing. Integrating PDLCO response with molecular data can reveal resistance mechanisms and aid in patient stratification for CAR T-cell therapy. Future plans include enhancing PDLCO complexity by introducing autologous immune cells, forming patient-derived immunocompetent lung cancer organoids. Our platform shows promise in informing and personalizing CAR T-cell therapy for lung cancer patients, potentially improving outcomes.