Personalized Modeling of CAR-T Cell Immunotherapy Using iPSC-Derived Effectors and Patient- Derived Tumor Organoids

Project Summary Chimeric antigen receptor (CAR) T cell therapy has transformed the treatment of hematologic malignancies but remains largely ineffective in solid tumors due to tumor heterogeneity, immune evasion, and the lack of physiologically relevant preclinical models. To address this gap, we propose to develop a personalized in vitro platform that combines tumor organoids derived from patient tumor tissues with CAR-T cells generated from induced pluripotent stem cells (iPSCs). This system will enable functional modeling of CAR-T cell activity, exhaustion, and tumor response in a patient-matched, autologous context. We will derive iPSCs from normal tissue adjacent to resected tumors, differentiate them into CD8+ T cells, and engineer them with tumor-targeting CARs. Tumor organoids from matched patients will be established through the Rutgers Cancer Institute Organoid Core, characterized for antigen expression and immunomodulatory features, and integrated into co-culture assays. Following co-culture, CAR-T cells will be evaluated for phenotype, cytotoxicity, cytokine production, and expression of exhaustion markers. The co-culture platform will allow real- time assessment of tumor cell killing, T cell activation/differentiation status, immune evasion, and microenvironmental interactions. This project addresses key goals of the NCI’s model development initiative by creating a scalable, autologous tumor-immune model for evaluating immunotherapies. The resulting platform will support translational discovery and the development of precision immunotherapies for solid tumors.