This study systematically evaluates the interactions between γδ T cells and patient-derived organoids through high-throughput single-cell analysis, revealing that multimodal cytotoxicity (AIC and ADCC) can overcome cancer cell immunomodulation and offering new therapeutic strategies for colorectal cancer, particularly the microsatellite-stable (MSS) type.
Literature Overview
This paper, titled 'Phenoscaping Reveals Multimodal γδ T-cell Cytotoxicity as a Strategy to Overcome Cancer Cell–Mediated Immunomodulation', published in the journal Cancer Research, reviews and summarizes the cytotoxic mechanisms of γδ T cells in various colorectal cancer organoid models and how their activity is influenced by cancer cell immunomodulation. Using systematic single-cell analysis, the study reveals functional heterogeneity of γδ T cells across different patient-derived organoids and explores how their anti-tumor activity can be enhanced through engineered modifications.
Background Knowledge
Colorectal cancer is a highly heterogeneous solid tumor, and its tumor microenvironment (TME) is typically immunosuppressive, especially in the microsatellite-stable (MSS) subtype, which shows poor response to immune checkpoint inhibitors. γδ T cells are innate immune cells capable of MHC-independent cytotoxicity and can also kill tumor cells via Fcγ receptor-mediated ADCC (Antibody-Dependent Cell-Mediated Cytotoxicity). However, the functional regulatory mechanisms of γδ T cells across different donors and tumor models are not fully understood. Moreover, colorectal cancer stem cells (CSCs) exhibit high plasticity, transitioning from chemotherapy-sensitive proCSCs to drug-resistant revCSCs, which complicates treatment. This study employs high-throughput single-cell analysis (TOBis MC) and organoid co-culture systems to systematically map the interaction landscape between γδ T cells and colorectal cancer organoids, providing a theoretical basis for developing more effective γδ T-cell immunotherapies.
Research Methods and Experiments
The research team employed a novel technology termed TOBis MC (Thiol-Reactive Organoid Barcoding in situ Mass Cytometry) to perform high-dimensional single-cell analysis on over 1,000 co-cultures of γδ T cells and patient-derived colorectal cancer organoids (PDOs). Using this approach, they quantified signal pathway activation, cell cycle status, apoptosis levels, and changes in immune phenotypes of γδ T cells. Additionally, CRISPR technology was used to generate B7-H3 knockout PDO models to assess whether ADCC functionality of γδ T cells is dependent on B7-H3 expression. Flow cytometry and single-cell RNA sequencing (scRNA-seq) were further employed to validate the immunomodulatory mechanisms of γδ T cells.
Key Conclusions and Perspectives
Research Significance and Prospects
This study reveals the multimodal cytotoxic mechanisms of γδ T cells in colorectal cancer therapy and demonstrates their ability to overcome cancer cell immunomodulation after engineering. Future research should focus on further optimizing engineered γδ T-cell designs, exploring their application in other solid tumors, and integrating personalized medicine strategies to improve anti-tumor efficacy.
Conclusion
Through high-throughput single-cell phenotypic analysis, this study systematically uncovered the complex interaction network between γδ T cells and colorectal cancer organoids. It was found that engineered γδ T cells expressing stIL15 can sustain cytotoxicity autonomously and overcome cancer cell immunosuppression through B7-H3-targeted ADCC. Additionally, the dual-mode (AIC + ADCC) killing strategy effectively eliminates chemotherapy-resistant cancer stem cells, offering a novel immunotherapeutic approach for colorectal cancer, especially the MSS subtype. These findings not only enhance our understanding of γδ T-cell anti-tumor mechanisms but also provide experimental support for optimizing γδ T-cell therapy in solid tumors.
