Vivier, E., Tomasello, E., Baratin, M., Walzer, T. & Ugolini, S. Functions of natural killer cells. Nat. Immunol. 9, 503–510 (2008).
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).
Google Scholar
Guillerey, C., Huntington, N. D. & Smyth, M. J. Targeting natural killer cells in cancer immunotherapy. Nat. Immunol. 17, 1025–1036 (2016).
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).
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).
Google Scholar
Crinier, A., Narni-Mancinelli, E., Ugolini, S. & Vivier, E. SnapShot: natural killer cells. Cell 180, 1280–1280 (2020).
Google Scholar
Lanier, L. L. Up on the tightrope: natural killer cell activation and inhibition. Nat. Immunol. 9, 495–502 (2008).
Google Scholar
Myers, J. A. & Miller, J. S. Exploring the NK cell platform for cancer immunotherapy. Nat. Rev. Clin. Oncol. 18, 85–100 (2021).
Google Scholar
Huntington, N. D., Cursons, J. & Rautela, J. The cancer–natural killer cell immunity cycle. Nat. Rev. Cancer 20, 437–454 (2020).
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).
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).
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).
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).
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).
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).
Google Scholar
Wrona, E., Borowiec, M. & Potemski, P. CAR-NK cells in the treatment of solid tumors. Int. J. Mol. Sci. 22, 5899 (2021).
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).
Google Scholar
Shimasaki, N., Jain, A. & Campana, D. NK cells for cancer immunotherapy. Nat. Rev. Drug Discov. 19, 200–218 (2020).
Google Scholar
Cozar, B. et al. Tumor-infiltrating natural killer cells. Cancer Discov. 11, 34–44 (2021).
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).
Google Scholar
Vivier, E. et al. Natural killer cell therapies. Nature 626, 727–736 (2024).
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).
Google Scholar
Delconte, R. B. et al. CIS is a potent checkpoint in NK cell-mediated tumor immunity. Nat. Immunol. 17, 816–824 (2016).
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).
Google Scholar
Delconte, R. B. et al. NK cell priming from endogenous homeostatic signals is modulated by CIS. Front. Immunol. 11, 75 (2020).
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).
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).
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).
Google Scholar
Qin, Y. et al. Developing enhanced immunotherapy using NKG2A knockout human pluripotent stem cell-derived NK cells. Cell Rep. 43, 114867 (2024).
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).
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).
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).
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).
Google Scholar
Zhang, X. et al. Gene knockout in cellular immunotherapy: application and limitations. Cancer Lett. 540, 215736 (2022).
Google Scholar
Konermann, S. et al. Genome-scale transcriptional activation by an engineered CRISPR–Cas9 complex. Nature 517, 583–588 (2015).
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).
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).
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).
Google Scholar
Patel, S. J. et al. Identification of essential genes for cancer immunotherapy. Nature 548, 537–542 (2017).
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).
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).
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).
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).
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).
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).
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).
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).
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).
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).
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).
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).
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).
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).
Google Scholar
Allen, F. et al. JACKS: joint analysis of CRISPR/Cas9 knockout screens. Genome Res. 29, 464–471 (2019).
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).
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).
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).
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).
Google Scholar
Jia, G., Wang, X. & Xiao, G. A permutation-based non-parametric analysis of CRISPR screen data. BMC Genomics 18, 545 (2017).
Google Scholar
Wang, T. et al. Identification and characterization of essential genes in the human genome. Science 350, 1096–1101 (2015).
Google Scholar
Wang, T. et al. Gene essentiality profiling reveals gene networks and synthetic lethal interactions with oncogenic Ras. Cell 168, 890–903 (2017).
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).
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).
Google Scholar
Hart, T. et al. High-resolution CRISPR screens reveal fitness genes and genotype-specific cancer liabilities. Cell 163, 1515–1526 (2015).
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).
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).
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).
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).
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).
Google Scholar
Frankish, A. et al. Gencode 2021. Nucleic Acids Res. 49, D916–D923 (2021).
Google Scholar
Hao, Y. et al. Dictionary learning for integrative, multimodal and scalable single-cell analysis. Nat. Biotechnol. 42, 293–304 (2024).
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).
Google Scholar
Tang, F. et al. A pan-cancer single-cell panorama of human natural killer cells. Cell 186, 4235–4251 (2023).
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).
Google Scholar
Aibar, S. et al. SCENIC: single-cell regulatory network inference and clustering. Nat. Methods 14, 1083–1086 (2017).
Google Scholar
Schaefer, C. F. et al. PID: the Pathway Interaction Database. Nucleic Acids Res. 37, D674–D679 (2009).
Google Scholar
Liberzon, A. et al. The Molecular Signatures Database (MSigDB) hallmark gene set collection. Cell Syst. 1, 417–425 (2015).
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).
Google Scholar

