Antigen-specific T cell receptor-engineered T cell (TCR-T) based immunotherapy has proven to be an effective method to combat cancer. In recent years, cross-talk between the innate and adaptive immune systems may be requisite to optimize sustained antigen-specific immunity, and the stimulator of interferon genes (STING) is a promising therapeutic target for cancer immunotherapy. The level of expression or presentation of antigen in tumor cells affects the recognition and killing of tumor cells by TCR-T. This study aimed at investigating the potential of innate immune stimulation of T cells and engineered T cells to enhance immunotherapy for low-expression antigen cancer cells. We systematically investigated the function and mechanism of cross-talk between STING agonist diABZI and adaptive immune systems. We established NY-ESO-1 full knockout Mel526 cells for this research and found that diABZI activated STING media and TCR signaling pathways. In addition, the results of flow cytometry showed that antigens presentation from cancer cells induced by STING agonist diABZI also improved the affinity of TCR-T cells function against tumor cells in vitro and in vivo. Our findings revealed that diABZI enhanced the immunotherapy efficacy of TCR-T by activating STING media and TCR signaling pathways, improving interferon-γ expression, and increasing antigens presentation of tumor cells. This indicates that STING agonist could be used as a strategy to promote TCR-T cancer immunotherapy.

Immunotherapies such as chimeric antigen receptor (CAR) T cells have shown promising outcomes in hematological cancer but face challenges in targeting solid tumors like glioblastoma (GBM). Advancing this therapy for GBM has been hindered by the lack of preclinical tools that accurately model the complex interplay between CAR T cells and tumor cells within the tumor microenvironment (TME) — interactions critical for optimizing CAR constructs and improving efficacy. Physiologically relevant models that closely mimic the solid TME are therefore highly sought after in developing CAR T therapies. Here, we report a microengineered glioblastoma-on-a-chip (GOC) model with a functional vascular network to investigate the efficacy and selectivity of IL-13 mutein CAR T cells (TV-13) against U87 GBM tumor cells expressing high interleukin-13 receptor alpha-2 (IL13Rα2), compared with the ubiquitously expressed IL13Rα1. This biomimetic platform recapitulates the GBM TME and enables dynamic evaluation of CAR T cell responses under locoregional administration, paralleling clinical approaches. Using the organotypic GOC model, we evaluated CAR T cell-mediated inhibition of GBM invasion, monitored real-time dynamic CAR T-U87 interactions, and quantified the release of cytotoxic, proinflammatory, and stimulation-associated cytokines as measures of T cell effector function. CAR T cells induced a density-dependent reduction in U87 migration, accompanied by robust cytokine release, while TV-13 maintained specificity towards IL13Rα2 tumor antigen over IL13Rα1. Additionally, we further demonstrated the efficacy of CAR T cells against patient-derived GBM cells within the GOC model. Collectively, these findings highlight the GOC platform as a powerful preclinical screening tool for cancer immunotherapy optimization.

Background: The dysregulation of the CDK1/Cyclin B1 complex drives tumor progression in breast cancer (BC). The natural flavonoid luteolin (LT) shows anti-cancer potential, but its mechanism targeting CDK1/CCNB1 remains unclear. Methods: CDK1, CCNB1, and CCNB2 expression were profiled in normal and BC cell lines. An engineered HEK293T GST-CDK1/CCNB1 cell model was used to evaluate LT’s effects on proliferation, ROS levels, and target gene transcription. Computational approaches (molecular docking, dynamics simulations, pharmacophore modeling, MM/GBSA, ADMET, and network pharmacology) assessed LT and its analogues. Results: CDK1/CCNB1 expression was lower in MCF7 BC cells than in normal cells, suggesting the loss of a growth barrier. In engineered HEK293T cells, LT suppressed CCNB1 transcription with minimal effect on CDK1 levels, correlating with anti-proliferative and ROS-modulating effects. Computational analyses confirmed stable LT binding to the CDK1/CCNB1 complex. Designed LT analogues showed improved binding and favorable ADMET profiles. Network pharmacology identified cell cycle regulation, particularly in BC stem cells, as the primary pathway targeted. Conclusions: LT and its analogues inhibit the CDK1/Cyclin B1 complex, revealing a dual mechanism that suppresses both tumor growth and BC stemness.