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Background Chimeric Antigen Receptor T-cell (CAR-T) therapy has transformed treatments for hematologic malignancies, but efficacy in solid tumors remains elusive due to the immunosuppressive and heterogeneous tumor microenvironment (TME). To address this problem, CanFinis Therapeutics has developed a proprietary ex vivo TME derived humanised platform to test effectiveness and specificity of new generation CAR-T cells on tumor cell killing under clinically relevant tumor conditions.Methods The TME platform incorporates key cellular, immune and stromal components derived from patient tumor, creating a humanized ex vivo system to test more than twenty five conditions including immune cell infiltration, tumor cell proliferation, special coordination of immune cells and tumor cells, persistence, tumor architecture and tumor cytotoxicity. A novel CAR-T product targeting metastatic gastric cancer was developed and validated using this system. Initially, in-vitro cytotoxicity assay (LDH release) was performed using the CAR-T cells, WT-T cells (Wild type T-cells) (effector) and HT29 cells (target). Subsequently, the generated CAR-T was tested for its ability to kill tumor cells in patient relevant TME with immune suppression, maintaining cytokine secretion profiles and hypoxia etc.Results The in-vitro cytotoxicity assay showed that CAR-T cells induced 32.6% cytotoxicity against HT29 cells over 24 hours, compared to 23% cytotoxicity induced by WT-T cells. This indicates that CAR-T cells exhibit greater target specificity and a more potent cytotoxic effect than WT-T cells. Comparable potency and sustained cytotoxic activity were exhibited by the CAR-Ts against tumor cells in the ex vivo TME setting. The CAR-T cells maintained effector function, produced key cytokines (e.g., IFN-γ, IL-2), and resisted cell exhaustion markers commonly induced by TME stressors. These findings confirm the efficacy of our CART cells even in presence of critical barriers to CAR-T efficacy in solid tumors settings. The product is now ready for GMP manufacturing and first-in-human clinical trials in metastatic gastric cancer in India.Conclusions The integration of CAR-T development with humanized TME platform provides a personalized and powerful preclinical strategy to assess the efficacy of CAR-T cells in solid tumor settings. These data support the advancement of a novel CAR-T therapy for metastatic gastric cancer and demonstrate the potential of TME relevant validation to accelerate CAR-T pipelines for other solid tumors.