OR7A10 GPCR engineering boosts CAR-NK therapy against solid tumours

OR7A10 GPCR engineering boosts CAR-NK therapy against solid tumours

  • Vivier, E., Tomasello, E., Baratin, M., Walzer, T. & Ugolini, S. Functions of natural killer cells. Nat. Immunol. 9, 503–510 (2008).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Ben-Shmuel, A., Biber, G. & Barda-Saad, M. Unleashing natural killer cells in the tumor microenvironment—the next generation of immunotherapy?. Front. Immunol. 11, 275 (2020).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Guillerey, C., Huntington, N. D. & Smyth, M. J. Targeting natural killer cells in cancer immunotherapy. Nat. Immunol. 17, 1025–1036 (2016).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Morvan, M. G. & Lanier, L. L. NK cells and cancer: you can teach innate cells new tricks. Nat. Rev. Cancer 16, 7–19 (2016).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Raulet, D. H. & Guerra, N. Oncogenic stress sensed by the immune system: role of natural killer cell receptors. Nat. Rev. Immunol. 9, 568–580 (2009).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Crinier, A., Narni-Mancinelli, E., Ugolini, S. & Vivier, E. SnapShot: natural killer cells. Cell 180, 1280–1280 (2020).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Lanier, L. L. Up on the tightrope: natural killer cell activation and inhibition. Nat. Immunol. 9, 495–502 (2008).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Myers, J. A. & Miller, J. S. Exploring the NK cell platform for cancer immunotherapy. Nat. Rev. Clin. Oncol. 18, 85–100 (2021).

    Article 
    PubMed 

    Google Scholar
     

  • Huntington, N. D., Cursons, J. & Rautela, J. The cancer–natural killer cell immunity cycle. Nat. Rev. Cancer 20, 437–454 (2020).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Bald, T., Krummel, M. F., Smyth, M. J. & Barry, K. C. The NK cell–cancer cycle: advances and new challenges in NK cell-based immunotherapies. Nat. Immunol. 21, 835–847 (2020).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Elahi, R., Heidary, A. H., Hadiloo, K. & Esmaeilzadeh, A. Chimeric antigen receptor-engineered natural killer (CAR NK) cells in cancer treatment; recent advances and future prospects. Stem Cell Rev. Rep. 17, 2081–2106 (2021).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Marofi, F. et al. Renaissance of armored immune effector cells, CAR-NK cells, brings the higher hope for successful cancer therapy. Stem Cell Res. Ther. 12, 200 (2021).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Dagher, O. K. & Posey, A. D. Jr. Forks in the road for CAR T and CAR NK cell cancer therapies. Nat. Immunol. 24, 1994–2007 (2023).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Hu, W., Wang, G., Huang, D., Sui, M. & Xu, Y. Cancer immunotherapy based on natural killer cells: current progress and new opportunities. Front. Immunol. 10, 1205 (2019).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Albinger, N., Hartmann, J. & Ullrich, E. Current status and perspective of CAR-T and CAR-NK cell therapy trials in Germany. Gene Ther. 28, 513–527 (2021).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Wrona, E., Borowiec, M. & Potemski, P. CAR-NK cells in the treatment of solid tumors. Int. J. Mol. Sci. 22, 5899 (2021).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Portillo, A. L. et al. Expanded human NK cells armed with CAR uncouple potent anti-tumor activity from off-tumor toxicity against solid tumors. iScience 24, 102619 (2021).

    Article 
    ADS 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Shimasaki, N., Jain, A. & Campana, D. NK cells for cancer immunotherapy. Nat. Rev. Drug Discov. 19, 200–218 (2020).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Cozar, B. et al. Tumor-infiltrating natural killer cells. Cancer Discov. 11, 34–44 (2021).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Christodoulou, I. et al. Engineering CAR-NK cells to secrete IL-15 sustains their anti-AML functionality but is associated with systemic toxicities. J. Immunother. Cancer 9, e003894 (2021).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Vivier, E. et al. Natural killer cell therapies. Nature 626, 727–736 (2024).

    Article 
    ADS 
    PubMed 
    CAS 

    Google Scholar
     

  • Bernard, P. L. et al. Targeting CISH enhances natural cytotoxicity receptor signaling and reduces NK cell exhaustion to improve solid tumor immunity. J. Immunother. Cancer 10, e004244 (2022).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Delconte, R. B. et al. CIS is a potent checkpoint in NK cell-mediated tumor immunity. Nat. Immunol. 17, 816–824 (2016).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Zhu, H. et al. Metabolic reprograming via deletion of CISH in human iPSC-derived NK cells promotes in vivo persistence and enhances anti-tumor activity. Cell Stem Cell 27, 224–237 (2020).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Delconte, R. B. et al. NK cell priming from endogenous homeostatic signals is modulated by CIS. Front. Immunol. 11, 75 (2020).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Nakazawa, T. et al. CIS deletion by CRISPR/Cas9 enhances human primary natural killer cell functions against allogeneic glioblastoma. J. Exp. Clin. Cancer Res. 42, 205 (2023).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Morimoto, T. et al. CRISPR–Cas9-Mediated TIM3 knockout in human natural killer cells enhances growth inhibitory effects on human glioma cells. Int. J. Mol. Sci. 22, 3489 (2021).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Mac Donald, A. et al. KLRC1 knockout overcomes HLA-E-mediated inhibition and improves NK cell antitumor activity against solid tumors. Front. Immunol. 14, 1231916 (2023).

    Article 
    CAS 

    Google Scholar
     

  • Qin, Y. et al. Developing enhanced immunotherapy using NKG2A knockout human pluripotent stem cell-derived NK cells. Cell Rep. 43, 114867 (2024).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Nakazawa, T. et al. Characterization of HIF-1α knockout primary human natural killer cells including populations in allogeneic glioblastoma. Int. J. Mol. Sci. 25, 5896 (2024).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Ni, J. et al. Single-cell RNA sequencing of tumor-infiltrating NK cells reveals that inhibition of transcription factor HIF-1α unleashes NK cell activity. Immunity 52, 1075–1087 (2020).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Peng, L. et al. In vivo AAV–SB-CRISPR screens of tumor-infiltrating primary NK cells identify genetic checkpoints of CAR-NK therapy. Nat. Biotechnol. 43, 752–761 (2025).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Rafei, H. et al. CREM is a regulatory checkpoint of CAR and IL-15 signalling in NK cells. Nature 643, 1076–1086 (2025).

    Article 
    ADS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Zhang, X. et al. Gene knockout in cellular immunotherapy: application and limitations. Cancer Lett. 540, 215736 (2022).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Konermann, S. et al. Genome-scale transcriptional activation by an engineered CRISPR–Cas9 complex. Nature 517, 583–588 (2015).

    Article 
    ADS 
    PubMed 
    CAS 

    Google Scholar
     

  • Zhou, X., Renauer, P. A., Zhou, L., Fang, S. Y. & Chen, S. Applications of CRISPR technology in cellular immunotherapy. Immunol. Rev. 320, 199–216 (2023).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Chen, H. et al. APLN promotes hepatocellular carcinoma through activating PI3K/Akt pathway and is a druggable target. Theranostics 9, 5246–5260 (2019).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Wang, Q. et al. APLN promotes the proliferation, migration, and glycolysis of cervical cancer through the PI3K/AKT/mTOR pathway. Arch. Biochem. Biophys. 755, 109983 (2024).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Patel, S. J. et al. Identification of essential genes for cancer immunotherapy. Nature 548, 537–542 (2017).

    Article 
    ADS 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Karagiota, A., Kanoura, A., Paraskeva, E., Simos, G. & Chachami, G. Pyruvate dehydrogenase phosphatase 1 (PDP1) stimulates HIF activity by supporting histone acetylation under hypoxia. FEBS J. 290, 2165–2179 (2023).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Song, Y., Zhang, J., Zhang, L., Zhang, S. & Shen, C. PDP1 promotes cell malignant behavior and is associated with worse clinical features in ovarian cancer patients: evidence from bioinformatics and in vitro level. Comput. Math. Methods Med. 2022, 7397250 (2022).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Yuan, M. et al. PDP1 promotes KRAS mutant colorectal cancer progression by serving as a scaffold for BRAF and MEK1. Cancer Lett. 597, 217007 (2024).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Alshamleh, I. et al. PDP1 is a key metabolic gatekeeper and modulator of drug resistance in FLT3-ITD-positive acute myeloid leukemia. Leukemia 37, 2367–2382 (2023).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Marin, D. et al. Safety, efficacy and determinants of response of allogeneic CD19-specific CAR-NK cells in CD19+ B cell tumors: a phase 1/2 trial. Nat. Med. 30, 772–784 (2024).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Kremer, V. et al. Genetic engineering of human NK cells to express CXCR2 improves migration to renal cell carcinoma. J. Immunother. Cancer 5, 73 (2017).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Sun, Y. et al. Adaptive NK cells exhibit tumor-specific immune memory and cytotoxicity in ovarian cancer. Cancer Immunol. Res. 13, 1080–1097 (2025).

  • Liu, E. et al. Cord blood NK cells engineered to express IL-15 and a CD19-targeted CAR show long-term persistence and potent antitumor activity. Leukemia 32, 520–531 (2018).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Liu, E. et al. Use of CAR-transduced natural killer cells in CD19-positive lymphoid tumors. N. Engl. J. Med. 382, 545–553 (2020).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Choi, C. & Finlay, D. K. Optimising NK cell metabolism to increase the efficacy of cancer immunotherapy. Stem Cell Res. Ther. 12, 320 (2021).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Egli, L. et al. CAR T cells outperform CAR NK cells in CAR-mediated effector functions in head-to-head comparison. Exp. Hematol. Oncol. 13, 51 (2024).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Modak, S., Kramer, K., Gultekin, S. H., Guo, H. F. & Cheung, N. K. Monoclonal antibody 8H9 targets a novel cell surface antigen expressed by a wide spectrum of human solid tumors. Cancer Res. 61, 4048–4054 (2001).

    PubMed 
    CAS 

    Google Scholar
     

  • Ahmed, M. et al. Humanized affinity-matured monoclonal antibody 8H9 has potent antitumor activity and binds to FG loop of tumor antigen B7-H3. J. Biol. Chem. 290, 30018–30029 (2015).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Ye, L. et al. In vivo CRISPR screening in CD8 T cells with AAV-Sleeping Beauty hybrid vectors identifies membrane targets for improving immunotherapy for glioblastoma. Nat. Biotechnol. 37, 1302–1313 (2019).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Langmead, B., Trapnell, C., Pop, M. & Salzberg, S. L. Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol. 10, R25 (2009).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Allen, F. et al. JACKS: joint analysis of CRISPR/Cas9 knockout screens. Genome Res. 29, 464–471 (2019).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Lun, A. T., Chen, Y. & Smyth, G. K. It’s DE-licious: a recipe for differential expression analyses of RNA-seq experiments using quasi-likelihood methods in edgeR. Methods Mol. Biol. 1418, 391–416 (2016).

    Article 
    PubMed 

    Google Scholar
     

  • Li, W. et al. MAGeCK enables robust identification of essential genes from genome-scale CRISPR/Cas9 knockout screens. Genome Biol. 15, 554 (2014).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Kim, E. & Hart, T. Improved analysis of CRISPR fitness screens and reduced off-target effects with the BAGEL2 gene essentiality classifier. Genome Med. 13, 2 (2021).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Jeong, H. H., Kim, S. Y., Rousseaux, M. W. C., Zoghbi, H. Y. & Liu, Z. Beta-binomial modeling of CRISPR pooled screen data identifies target genes with greater sensitivity and fewer false negatives. Genome Res. 29, 999–1008 (2019).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Jia, G., Wang, X. & Xiao, G. A permutation-based non-parametric analysis of CRISPR screen data. BMC Genomics 18, 545 (2017).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Wang, T. et al. Identification and characterization of essential genes in the human genome. Science 350, 1096–1101 (2015).

    Article 
    ADS 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Wang, T. et al. Gene essentiality profiling reveals gene networks and synthetic lethal interactions with oncogenic Ras. Cell 168, 890–903 (2017).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Aguirre, A. J. et al. Genomic copy number dictates a gene-independent cell response to CRISPR/Cas9 targeting. Cancer Discov. 6, 914–929 (2016).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Meyers, R. M. et al. Computational correction of copy number effect improves specificity of CRISPR–Cas9 essentiality screens in cancer cells. Nat. Genet. 49, 1779–1784 (2017).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Hart, T. et al. High-resolution CRISPR screens reveal fitness genes and genotype-specific cancer liabilities. Cell 163, 1515–1526 (2015).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Sullivan, R. J. et al. First-in-class ERK1/2 inhibitor ulixertinib (BVD-523) in patients with MAPK mutant advanced solid tumors: results of a Phase I dose-escalation and expansion study. Cancer Discov. 8, 184–195 (2018).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Lei, W. et al. Safety and feasibility of 4-1BB co-stimulated CD19-specific CAR-NK cell therapy in refractory/relapsed large B cell lymphoma: a phase 1 trial. Nat. Cancer 6, 786–800 (2025).

    Article 
    ADS 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Albinger, N. et al. Primary CD33-targeting CAR-NK cells for the treatment of acute myeloid leukemia. Blood Cancer J. 12, 61 (2022).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Guo, S. et al. CD70-specific CAR NK cells expressing IL-15 for the treatment of CD19-negative B-cell malignancy. Blood Adv. 8, 2635–2645 (2024).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Bexte, T. et al. CRISPR/Cas9 editing of NKG2A improves the efficacy of primary CD33-directed chimeric antigen receptor natural killer cells. Nat. Commun. 15, 8439 (2024).

    Article 
    ADS 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Frankish, A. et al. Gencode 2021. Nucleic Acids Res. 49, D916–D923 (2021).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Hao, Y. et al. Dictionary learning for integrative, multimodal and scalable single-cell analysis. Nat. Biotechnol. 42, 293–304 (2024).

    Article 
    ADS 
    PubMed 
    CAS 

    Google Scholar
     

  • Welch, J. D. et al. Single-cell multi-omic integration compares and contrasts features of brain cell identity. Cell 177, 1873–1887 (2019).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Tang, F. et al. A pan-cancer single-cell panorama of human natural killer cells. Cell 186, 4235–4251 (2023).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Bergen, V., Lange, M., Peidli, S., Wolf, F. A. & Theis, F. J. Generalizing RNA velocity to transient cell states through dynamical modeling. Nat. Biotechnol. 38, 1408–1414 (2020).

    Article 
    ADS 
    PubMed 
    CAS 

    Google Scholar
     

  • Aibar, S. et al. SCENIC: single-cell regulatory network inference and clustering. Nat. Methods 14, 1083–1086 (2017).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Schaefer, C. F. et al. PID: the Pathway Interaction Database. Nucleic Acids Res. 37, D674–D679 (2009).

    Article 
    PubMed 
    CAS 

    Google Scholar
     

  • Liberzon, A. et al. The Molecular Signatures Database (MSigDB) hallmark gene set collection. Cell Syst. 1, 417–425 (2015).

    Article 
    PubMed 
    PubMed Central 
    CAS 

    Google Scholar
     

  • Rosskopf, S. et al. A Jurkat 76 based triple parameter reporter system to evaluate TCR functions and adoptive T cell strategies. Oncotarget 9, 17608–17619 (2018).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar