This study explores the therapeutic potential of engineered induced pluripotent stem cell-derived natural killer (iNK) cells targeting the TIGIT-CD155 axis and CD73 in glioblastoma (GBM). The authors developed a synNotch-based construct that activates iNK cells upon binding to CD155, leading to the release of an anti-CD73 single-chain variable fragment (scFv) to block purinergic activity and enhance NK cell cytotoxicity. Key findings include: 1. **CD155 and CD73 Expression and Prognosis**: CD155 and CD73 are highly expressed in GBM and act as negative prognostic factors, with co-expression of both genes being a significant indicator of poor patient outcomes. 2. **Engineered iNK Cells**: iNK cells engineered with the synNotch construct were successfully generated and showed similar phenotypic markers to non-engineered iNK cells, maintaining high purity and viability during differentiation and expansion. 3. **Dual Targeting of CD73 and CD155**: Engineered iNK cells targeting both CD73 and CD155 demonstrated enhanced cytotoxicity against GBM cells compared to single-targeting or control groups, indicating the importance of dual blockade in overcoming immunosuppressive mechanisms. 4. **In Vivo Efficacy**: In immunocompetent mouse models, dual targeting of CD73 and CD155 with engineered iNK cells significantly slowed tumor growth, increased the infiltration of NK and CD8+ T cells, and reduced M2 macrophages, leading to improved survival rates. 5. **Orthotopic Patient-Derived Xenograft Model**: In an orthotopic GBM model, engineered iNK cells showed dramatic reduction in tumor growth and improved survival, further validating the therapeutic potential of this approach. 6. **Discussion**: The study highlights the effectiveness of dual targeting of CD73 and CD155 in overcoming immunosuppressive mechanisms in GBM, suggesting that this approach could be a promising therapeutic strategy for treating this aggressive tumor. Overall, the study demonstrates the potential of synNotch-engineered iNK cells as a powerful therapeutic platform for GBM, offering a novel approach to combat the heterogeneity and immunosuppressive nature of this disease.This study explores the therapeutic potential of engineered induced pluripotent stem cell-derived natural killer (iNK) cells targeting the TIGIT-CD155 axis and CD73 in glioblastoma (GBM). The authors developed a synNotch-based construct that activates iNK cells upon binding to CD155, leading to the release of an anti-CD73 single-chain variable fragment (scFv) to block purinergic activity and enhance NK cell cytotoxicity. Key findings include: 1. **CD155 and CD73 Expression and Prognosis**: CD155 and CD73 are highly expressed in GBM and act as negative prognostic factors, with co-expression of both genes being a significant indicator of poor patient outcomes. 2. **Engineered iNK Cells**: iNK cells engineered with the synNotch construct were successfully generated and showed similar phenotypic markers to non-engineered iNK cells, maintaining high purity and viability during differentiation and expansion. 3. **Dual Targeting of CD73 and CD155**: Engineered iNK cells targeting both CD73 and CD155 demonstrated enhanced cytotoxicity against GBM cells compared to single-targeting or control groups, indicating the importance of dual blockade in overcoming immunosuppressive mechanisms. 4. **In Vivo Efficacy**: In immunocompetent mouse models, dual targeting of CD73 and CD155 with engineered iNK cells significantly slowed tumor growth, increased the infiltration of NK and CD8+ T cells, and reduced M2 macrophages, leading to improved survival rates. 5. **Orthotopic Patient-Derived Xenograft Model**: In an orthotopic GBM model, engineered iNK cells showed dramatic reduction in tumor growth and improved survival, further validating the therapeutic potential of this approach. 6. **Discussion**: The study highlights the effectiveness of dual targeting of CD73 and CD155 in overcoming immunosuppressive mechanisms in GBM, suggesting that this approach could be a promising therapeutic strategy for treating this aggressive tumor. Overall, the study demonstrates the potential of synNotch-engineered iNK cells as a powerful therapeutic platform for GBM, offering a novel approach to combat the heterogeneity and immunosuppressive nature of this disease.