[1] Wolf N K, Kissiov D U, Raulet D H. Roles of natural killer cells in immunity to cancer, and applications to immunotherapy [J]. Nature Reviews Immunology, 2023, 23(2): 90-105.
[2] Alfarra H, Weir J, Grieve S, et al. Targeting NK cell inhibitory receptors for precision multiple myeloma immunotherapy [J]. Frontiers in Immunology, 2020, 11: 575609.
[3] Pallmer K, Oxenius A. Recognition and regulation of T cells by NK cells [J]. Frontiers in Immunology, 2016, 7: 251.
[4] Coënon L, Geindreau M, Ghiringhelli F, et al. Natural Killer cells at the frontline in the fight against cancer [J]. Cell Death & Disease, 2024, 15: 614.
[5] Ma S B, Yu J H, Caligiuri M A. Natural killer cell–based immunotherapy for cancer [J]. The Journal of Immunology, 2025, 214(7): 1444-1456.
[6] Zhang A Q, Yang X C, Zhang Y C, et al. Unlocking the potential of CAR-NK cell therapy: Overcoming barriers and challenges in the treatment of myeloid malignancies [J]. Molecular Cancer Therapeutics, 2025, 24(4): 536-549.
[7] Fujisaki H, Kakuda H, Shimasaki N, et al. Expansion of highly cytotoxic human natural killer cells for cancer cell therapy [J]. Cancer Research, 2009, 69(9): 4010-4017.
[8] Grzywacz B, Moench L, McKenna D, et al. Natural killer cell homing and persistence in the bone marrow after adoptive immunotherapy correlates with better leukemia control [J]. Journal of Immunotherapy, 2019, 42(2): 65-72.
[9] Björklund A T, Carlsten M, Sohlberg E, et al. Complete remission with reduction of high-risk clones following haploidentical NK-cell therapy against MDS and AML [J]. Clinical Cancer Research, 2018, 24(8): 1834-1844.
[10] Denman C J, Senyukov V V, Somanchi S S, et al. Membrane-bound IL-21 promotes sustained ex vivo proliferation of human natural killer cells [J]. PLoS One, 2012, 7(1): e30264.
[11] Zhang D, Zheng Y S, Lin Z G, et al. Equipping natural killer cells with specific targeting and checkpoint blocking aptamers for enhanced adoptive immunotherapy in solid tumors [J]. Angewandte Chemie International Edition, 2020, 59(29): 12022-12028.
[12] Murray S, Lundqvist A. Targeting the tumor microenvironment to improve natural killer cell-based immunotherapies: On being in the right place at the right time, with resilience [J]. Human Vaccines & Immunotherapeutics, 2016, 12(3): 607-611.
[13] Lu C G, Guo C J, Chen H, et al. A novel chimeric PD1-NKG2D-41BB receptor enhances antitumor activity of NK92 cells against human lung cancer H1299 cells by triggering pyroptosis [J]. Molecular Immunology, 2020, 122: 200-206.
[14] Li T X, Yang Y Y, Qi H Z, et al. CRISPR/Cas9 therapeutics: Progress and prospects [J]. Signal Transduction and Targeted Therapy, 2023, 8: 36.
[15] Li T Y, Li S Q, Kang Y, et al. Harnessing the evolving CRISPR/Cas9 for precision oncology [J]. Journal of Translational Medicine, 2024, 22(1): 749.
[16] Jinek M, Chylinski K, Fonfara I, et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity [J]. Science, 2012, 337(6096): 816-821.
[17] Zhang X H, Tee L Y, Wang X G, et al. Off-target effects in CRISPR/Cas9-mediated genome engineering [J]. Molecular Therapy - Nucleic Acids, 2015, 4: e264.
[18] Bhardwaj A, Nain V. TALENs: An indispensable tool in the era of CRISPR: A mini review [J]. Journal of Genetic Engineering and Biotechnology, 2021, 19(1): 125.
[19] Kampmann M. CRISPR-based functional genomics for neurological disease [J]. Nature Reviews Neurology, 2020, 16(9): 465-480.
[20] Kanafi M M, Tavallaei M. Overview of advances in CRISPR/deadCas9 technology and its applications in human diseases [J]. Gene, 2022, 830: 146518.
[21] Enright A L, Heelan W J, Ward R D, et al. CRISPRi functional genomics in bacteria and its application to medical and industrial research [J]. Microbiology and Molecular Biology Reviews, 2024, 88(2): e00170-22.
[22] Villegas Kcam M C, Tsong A J, Chappell J. Uncovering the distinct properties of a bacterial type I-E CRISPR activation system [J]. ACS Synthetic Biology, 2022, 11(2): 1000-1003.
[23] Cui X L, Zhang C, Xu Z F, et al. Dual CRISPR interference and activation for targeted reactivation of X-linked endogenous FOXP3 in human breast cancer cells [J]. Molecular Cancer, 2022, 21(1): 38.
[24] Gaudelli N M, Komor A C, Rees H A, et al. Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage [J]. Nature, 2017, 551(7681): 464-471.
[25] Anzalone A V, Randolph P B, Davis J R, et al. Search-and-replace genome editing without double-strand breaks or donor DNA [J]. Nature, 2019, 576(7785): 149-157.
[26] Chen P J, Liu D R. Prime editing for precise and highly versatile genome manipulation [J]. Nature Reviews Genetics, 2023, 24(3): 161-177.
[27] Gehrke J M, Cervantes O, Clement M K, et al. An APOBEC3A-Cas9 base editor with minimized bystander and off-target activities [J]. Nature Biotechnology, 2018, 36(10): 977-982.
[28] Kwon J, Kim M, Bae S, et al. TAPE-seq is a cell-based method for predicting genome-wide off-target effects of prime editor [J]. Nature Communications, 2022, 13: 7975.
[29] Liang S Q, Liu P P, Ponnienselvan K, et al. Genome-wide profiling of prime editor off-target sites in vitro and in vivo using PE-tag [J]. Nature Methods, 2023, 20(6): 898-907.
[30] Yu Z X, Lu Z K, Li J J, et al. PEAC-seq adopts Prime Editor to detect CRISPR off-target and DNA translocation [J]. Nature Communications, 2022, 13: 7545.
[31] González Castro N, Bjelic J, Malhotra G, et al. Comparison of the feasibility, efficiency, and safety of genome editing technologies [J]. International Journal of Molecular Sciences, 2021, 22(19): 10355.
[32] Chou S T, Leng Q X, Mixson A J. Zinc finger nucleases: Tailor-made for gene therapy [J]. Drugs of the Future, 2012, 37(3): 183-196.
[33] Gupta D, Bhattacharjee O, Mandal D, et al. CRISPR-Cas9 system: A new-fangled dawn in gene editing [J]. Life Sciences, 2019, 232: 116636.
[34] Tang S Y, Zha S J, Du Z C, et al. Targeted integration of EpCAM-specific CAR in human induced pluripotent stem cells and their differentiation into NK cells [J]. Stem Cell Research & Therapy, 2021, 12(1): 580.
[35] Richter A, Streubel J, Boch J. TAL effector DNA-binding principles and specificity [J]. Methods in Molecular Biology, 2016, 1338: 9-25.
[36] Deng D, Yan C Y, Pan X J, et al. Structural basis for sequence-specific recognition of DNA by TAL effectors [J]. Science, 2012, 335(6069): 720-723.
[37] Li H Y, Yang Y, Hong W Q, et al. Applications of genome editing technology in the targeted therapy of human diseases: Mechanisms, advances and prospects [J]. Signal Transduction and Targeted Therapy, 2020, 5: 1.
[38] Chen A P, Gao P, Ashok P, et al. TALEN®-based gene edited iPSC-derived NK (iNK) cells demonstrate enhanced antitumor activity [J]. Journal for ImmunoTherapy of Cancer, 2022, 10: 323.
[39] Vargas J E, Chicaybam L, Stein R T, et al. Retroviral vectors and transposons for stable gene therapy: Advances, current challenges and perspectives [J]. Journal of Translational Medicine, 2016, 14(1): 288.
[40] Geurts A M, Yang Y, Clark K J, et al. Gene transfer into genomes of human cells by the sleeping beauty transposon system [J]. Molecular Therapy, 2003, 8(1): 108-117.
[41] Turchiano G, Latella M C, Gogol-Döring A, et al. Genomic analysis of Sleeping Beauty transposon integration in human somatic cells [J]. PLoS One, 2014, 9(11): e112712.
[42] Bire S, Casteret S, Arnaoty A, et al. Transposase concentration controls transposition activity: Myth or reality? [J]. Gene, 2013, 530(2): 165-171.
[43] Izsvák Z, Ivics Z. Sleeping beauty transposition: Biology and applications for molecular therapy [J]. Molecular Therapy, 2004, 9(2): 147-156.
[44] Ding S, Wu X H, Li G, et al. Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice [J]. Cell, 2005, 122(3): 473-483.
[45] Li M A, Turner D J, Ning Z M, et al. Mobilization of giant piggyBac transposons in the mouse genome [J]. Nucleic Acids Research, 2011, 39(22): e148.
[46] Cadiñanos J, Bradley A. Generation of an inducible and optimized piggyBac transposon system [J]. Nucleic Acids Research, 2007, 35(12): e87.
[47] Sun Y, Liu G, Huang Y. Applications of piggyBac transposons for genome manipulation in stem cells [J]. Stem Cells International, 2021, 2021(1): 3829286.
[48] Li M D, Bronson D L, Lemke T D, et al. Phylogenetic analyses of 55 retroelements on the basis of the nucleotide and product amino acid sequences of the pol gene [J]. Molecular Biology and Evolution, 1995, 12(4): 657-670.
[49] Fang M G, Allen A, Luo C, et al. Unlocking the potential of iPSC-derived immune cells: Engineering iNK and iT cells for cutting-edge immunotherapy [J]. Frontiers in Immunology, 2024, 15: 1457629.
[50] Goldenson B H, Hor P, Kaufman D S. iPSC-derived natural killer cell therapies - expansion and targeting [J]. Frontiers in Immunology, 2022, 13: 841107.
[51] Knorr D A, Ni Z Y, Hermanson D, et al. Clinical-scale derivation of natural killer cells from human pluripotent stem cells for cancer therapy [J]. Stem Cells Translational Medicine, 2013, 2(4): 274-283.
[52] Woll P S, Martin C H, Miller J S, et al. Human embryonic stem cell-derived NK cells acquire functional receptors and cytolytic activity [J]. Journal of Immunology, 2005, 175(8): 5095-5103.
[53] Wang X B, Zhang Y, Jin Y, et al. An iPSC-derived CD19/BCMA CAR-NK therapy in a patient with systemic sclerosis [J]. Cell, 2025, 188(16): 4225-4238.e12.
[54] Wu X Y, Matosevic S. Gene-edited and CAR-NK cells: Opportunities and challenges with engineering of NK cells for immunotherapy [J]. Molecular Therapy - Oncolytics, 2022, 27: 224-238.
[55] Müller S, Bexte T, Gebel V, et al. High cytotoxic efficiency of lentivirally and alpharetrovirally engineered CD19-specific chimeric antigen receptor natural killer cells against acute lymphoblastic leukemia [J]. Frontiers in Immunology, 2020, 10: 3123.
[56] Leivas A, Valeri A, Córdoba L, et al. NKG2D-CAR-transduced natural killer cells efficiently target multiple myeloma [J]. Blood Cancer Journal, 2021, 11: 146.
[57] Liu E L, Marin D, Banerjee P, et al. Use of CAR-transduced natural killer cells in CD19-positive lymphoid tumors [J]. New England Journal of Medicine, 2020, 382(6): 545-553.
[58] Marin D, Li Y, Basar R, 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 [J]. Nature Medicine, 2024, 30(3): 772-784.
[59] Bexte T, Albinger N, Al Ajami A, et al. CRISPR/Cas9 editing of NKG2A improves the efficacy of primary CD33-directed chimeric antigen receptor natural killer cells [J]. Nature Communications, 2024, 15: 8439.
[60] Albinger N, Pfeifer R, Nitsche M, et al. Primary CD33-targeting CAR-NK cells for the treatment of acute myeloid leukemia [J]. Blood Cancer Journal, 2022, 12: 61.
[61] Bexte T, Alzubi J, Reindl L M, et al. CRISPR-Cas9 based gene editing of the immune checkpoint NKG2A enhances NK cell mediated cytotoxicity against multiple myeloma [J]. OncoImmunology, 2022, 11(1): 2081415.
[62] Lin Y Q, Xiao Z Y, Hu F X, et al. Engineered CRO-CD7 CAR-NK cells derived from pluripotent stem cells avoid fratricide and efficiently suppress human T-cell malignancies [J]. Journal of Hematology & Oncology, 2025, 18(1): 57.
[63] Gomes-Silva D, Srinivasan M, Sharma S, et al. CD7-edited T cells expressing a CD7-specific CAR for the therapy of T-cell malignancies [J]. Blood, 2017, 130(3): 285-296.
[64] Peng L, Renauer P A, Sferruzza G, et al. In vivo AAV–SB-CRISPR screens of tumor-infiltrating primary NK cells identify genetic checkpoints of CAR-NK therapy [J]. Nature Biotechnology, 2025, 43(5): 752-761.
[65] Vivier E, Raulet D H, Moretta A, et al. Innate or adaptive immunity? The example of natural killer cells [J]. Science, 2011, 331(6013): 44-49.
[66] Floros T, Tarhini A A. Anticancer cytokines: Biology and clinical effects of interferon-α2, interleukin (IL)-2, IL-15, IL-21, and IL-12 [J]. Seminars in Oncology, 2015, 42(4): 539-548.
[67] Nagashima S, Mailliard R, Kashii Y, et al. Stable transduction of the interleukin-2 gene into human natural killer cell lines and their phenotypic and functional characterization in vitro and in vivo [J]. Blood, 1998, 91(10): 3850-3861.
[68] Ranson T, Vosshenrich C A J, Corcuff E, et al. IL-15 is an essential mediator of peripheral NK-cell homeostasis [J]. Blood, 2003, 101(12): 4887-4893.
[69] Wrangle J M, Velcheti V, Patel M R, et al. ALT-803, an IL-15 superagonist, in combination with nivolumab in patients with metastatic non-small cell lung cancer: A non-randomised, open-label, phase 1b trial [J]. The Lancet Oncology, 2018, 19(5): 694-704.
[70] Liu E, Tong Y, Dotti G, et al. Cord blood NK cells engineered to express IL-15 and a CD19-targeted CAR show long-term persistence and potent antitumor activity [J]. Leukemia, 2018, 32(2): 520-531.
[71] Xu X D, Cao P Y, Wang M, et al. Signaling intact membrane-bound IL-15 enables potent anti-tumor activity and safety of CAR-NK cells [J]. Frontiers in Immunology, 2025, 16: 1658580.
[72] Du Z C, Ng Y Y, Zha S J, et al. piggyBac system to co-express NKG2D CAR and IL-15 to augment the in vivo persistence and anti-AML activity of human peripheral blood NK cells [J]. Molecular Therapy - Methods & Clinical Development, 2021, 23: 582-596.
[73] Feng D D, Sun L, Hu D X, et al. Expression of membrane-bound Interleukin-15 sustains the growth and survival of CAR-NK cells [J]. International Immunopharmacology, 2025, 166: 115577.
[74] Miller J S, Morishima C, McNeel D G, et al. A first-in-human phase I study of subcutaneous outpatient recombinant human IL15 (rhIL15) in adults with advanced solid tumors [J]. Clinical Cancer Research, 2018, 24(7): 1525-1535.
[75] Jamali A, Hadjati J, Madjd Z, et al. Highly efficient generation of transgenically augmented CAR NK cells overexpressing CXCR4 [J]. Frontiers in Immunology, 2020, 11: 2028.
[76] Schomer N T, Jiang Z K, Lloyd M I, et al. CCR7 expression in CD19 chimeric antigen receptor-engineered natural killer cells improves migration toward CCL19-expressing lymphoma cells and increases tumor control in mice with human lymphoma [J]. Cytotherapy, 2022, 24(8): 827-834.
[77] Feigl F F, Stahringer A, Peindl M, et al. Efficient redirection of NK cells by genetic modification with chemokine receptors CCR4 and CCR2B [J]. International Journal of Molecular Sciences, 2023, 24(4): 3129.
[78] Lee N, Llano M, Carretero M, et al. HLA-E is a major ligand for the natural killer inhibitory receptor CD94/NKG2A [J]. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95(9): 5199-5204.
[79] André P, Denis C, Soulas C, et al. Anti-NKG2A MAb is a checkpoint inhibitor that promotes anti-tumor immunity by unleashing both T and NK cells [J]. Cell, 2018, 175(7): 1731-1743.e13.
[80] McWilliams E M, Mele J M, Cheney C, et al. Therapeutic CD94/NKG2A blockade improves natural killer cell dysfunction in chronic lymphocytic leukemia [J]. OncoImmunology, 2016, 5(10): e1226720.
[81] Cózar B, Greppi M, Carpentier S, et al. Tumor-infiltrating natural killer cells [J]. Cancer Discovery, 2021, 11(1): 34-44.
[82] Kamiya T, Seow S V, Wong D, et al. Blocking expression of inhibitory receptor NKG2A overcomes tumor resistance to NK cells [J]. Journal of Clinical Investigation, 2019, 129(5): 2094-2106.
[83] da Silva I P, Gallois A, Jimenez-Baranda S, et al. Reversal of NK-cell exhaustion in advanced melanoma by tim-3 blockade [J]. Cancer Immunology Research, 2014, 2(5): 410-422.
[84] Xu L Y, Huang Y Y, Tan L L, et al. Increased Tim-3 expression in peripheral NK cells predicts a poorer prognosis and Tim-3 blockade improves NK cell-mediated cytotoxicity in human lung adenocarcinoma [J]. International Immunopharmacology, 2015, 29(2): 635-641.
[85] Wang Z X, Zhu J L, Gu H D, et al. The clinical significance of abnormal tim-3 expression on NK cells from patients with gastric cancer [J]. Immunological Investigations, 2015, 44(6): 578-589.
[86] Sakuishi K, Apetoh L, Sullivan J M, et al. Targeting Tim-3 and PD-1 pathways to reverse T cell exhaustion and restore anti-tumor immunity [J]. Journal of Experimental Medicine, 2010, 207(10): 2187-2194.
[87] Pu F F, Chen F X, Zhang Z C, et al. TIM-3 expression and its association with overall survival in primary osteosarcoma [J]. Oncology Letters, 2019, 18(5): 5294-5300.
[88] Gallois A, Silva I, Osman I, et al. Reversal of natural killer cell exhaustion by TIM-3 blockade [J]. OncoImmunology, 2014, 3(12): e946365.
[89] Hou H Y, Liu W Y, Wu S J, et al. Tim-3 negatively mediates natural killer cell function in LPS-induced endotoxic shock [J]. PLoS One, 2014, 9(10): e110585.
[90] Zhang Q, Bi J C, Zheng X D, et al. Blockade of the checkpoint receptor TIGIT prevents NK cell exhaustion and elicits potent anti-tumor immunity [J]. Nature Immunology, 2018, 19(7): 723-732.
[91] Masson D, Jarry A, Baury B, et al. Overexpression of the CD155 gene in human colorectal carcinoma [J]. Gut, 2001, 49(2): 236-240.
[92] Li M, Xia P Y, Du Y, et al. T-cell immunoglobulin and ITIM domain (TIGIT) receptor/poliovirus receptor (PVR) ligand engagement suppresses interferon-γ production of natural killer cells via β-arrestin 2-mediated negative signaling [J]. Journal of Biological Chemistry, 2014, 289(25): 17647-17657.
[93] Carlsten M, Norell H, Bryceson Y T, et al. Primary human tumor cells expressing CD155 impair tumor targeting by down-regulating DNAM-1 on NK cells [J]. The Journal of Immunology, 2009, 183(8): 4921-4930.
[94] Sarhan D, Cichocki F, Zhang B, et al. Adaptive NK cells with low TIGIT expression are inherently resistant to myeloid-derived suppressor cells [J]. Cancer Research, 2016, 76(19): 5696-5706.
[95] Dong H D, Strome S E, Salomao D R, et al. Tumor-associated B7-H1 promotes T-cell apoptosis: A potential mechanism of immune evasion [J]. Nature Medicine, 2002, 8(8): 793-800.
[96] Pesce S, Greppi M, Grossi F, et al. PD/1-PD-ls checkpoint: Insight on the potential role of NK cells [J]. Frontiers in Immunology, 2019, 10: 1242.
[97] Quatrini L, Vacca P, Tumino N, et al. Glucocorticoids and the cytokines IL-12, IL-15, and IL-18 present in the tumor microenvironment induce PD-1 expression on human natural killer cells [J]. Journal of Allergy and Clinical Immunology, 2021, 147(1): 349-360.
[98] Dong Y N, Sun Q, Zhang X W. PD-1 and its ligands are important immune checkpoints in cancer [J]. Oncotarget, 2017, 8(2): 2171-2186.
[99] Benson D M Jr, Bakan C E, Mishra A, et al. The PD-1/PD-L1 axis modulates the natural killer cell versus multiple myeloma effect: A therapeutic target for CT-011, a novel monoclonal anti–PD-1 antibody [J]. Blood, 2010, 116(13): 2286-2294.
[100] Vari F, Arpon D, Keane C, et al. Immune evasion via PD-1/PD-L1 on NK cells and monocyte/macrophages is more prominent in Hodgkin lymphoma than DLBCL [J]. Blood, 2018, 131(16): 1809-1819.
[101] Poznanski S M, Singh K, Ritchie T M, et al. Metabolic flexibility determines human NK cell functional fate in the tumor microenvironment [J]. Cell Metabolism, 2021, 33(6): 1205-1220.e5.
[102] Husain Z, Huang Y N, Seth P, et al. Tumor-derived lactate modifies antitumor immune response: Effect on myeloid-derived suppressor cells and NK cells [J]. The Journal of Immunology, 2013, 191(3): 1486-1495.
[103] Zheng X H, Qian Y B, Fu B Q, et al. Mitochondrial fragmentation limits NK cell-based tumor immunosurveillance [J]. Nature Immunology, 2019, 20(12): 1656-1667.
[104] Allard B, Longhi M S, Robson S C, et al. The ectonucleotidases CD39 and CD73: Novel checkpoint inhibitor targets [J]. Immunological Reviews, 2017, 276(1): 121-144.
[105] Chambers A M, Wang J, Lupo K B, et al. Adenosinergic signaling alters natural killer cell functional responses [J]. Frontiers in Immunology, 2018, 9: 2533.
[106] Pataskar A, Champagne J, Nagel R, et al. Tryptophan depletion results in tryptophan-to-phenylalanine substitutants [J]. Nature, 2022, 603(7902): 721-727.
[107] Della Chiesa M, Carlomagno S, Frumento G, et al. The tryptophan catabolite l-kynurenine inhibits the surface expression of NKp46- and NKG2D-activating receptors and regulates NK-cell function [J]. Blood, 2006, 108(13): 4118-4125.
[108] Michelet X, Dyck L, Hogan A, et al. Metabolic reprogramming of natural killer cells in obesity limits antitumor responses [J]. Nature Immunology, 2018, 19(12): 1330-1340.
[109] O’Brien K L, Finlay D K. Immunometabolism and natural killer cell responses [J]. Nature Reviews Immunology, 2019, 19(5): 282-290.
[110] Ciurea S O, Schafer J R, Bassett R, et al. Phase 1 clinical trial using mbIL21 ex vivo–expanded donor-derived NK cells after haploidentical transplantation [J]. Blood, 2017, 130(16): 1857-1868.
[111] Viel S, Marçais A, Guimaraes F S, et al. TGF-β inhibits the activation and functions of NK cells by repressing the mTOR pathway [J]. Science Signaling, 2016, 9(415): ra19.
[112] Zaiatz-Bittencourt V, Finlay D K, Gardiner C M. Canonical TGF-β signaling pathway represses human NK cell metabolism [J]. The Journal of Immunology, 2018, 200(12): 3934-3941.
[113] Yvon E S, Burga R, Powell A, et al. Cord blood natural killer cells expressing a dominant negative TGF-β receptor: Implications for adoptive immunotherapy for glioblastoma [J]. Cytotherapy, 2017, 19(3): 408-418.
[114] Shaim H, Shanley M, Basar R, et al. Targeting the αv integrin/TGF-β axis improves natural killer cell function against glioblastoma stem cells [J]. Journal of Clinical Investigation, 2021, 131(14): e142116.
[115] Kim T D, Lee S U, Yun S, et al. Human microRNA-27a* targets Prf1 and GzmB expression to regulate NK-cell cytotoxicity [J]. Blood, 2011, 118(20): 5476-5486.
[116] Delconte R B, Kolesnik T B, Dagley L F, et al. CIS is a potent checkpoint in NK cell–mediated tumor immunity [J]. Nature Immunology, 2016, 17(7): 816-824.
[117] Bernard P L, Delconte R, Pastor S, et al. Targeting CISH enhances natural cytotoxicity receptor signaling and reduces NK cell exhaustion to improve solid tumor immunity [J]. Journal for ImmunoTherapy of Cancer, 2022, 10(5): e004244.
[118] Zhu H, Blum R H, Bernareggi D, et al. Metabolic reprograming via deletion of CISH in human iPSC-derived NK cells promotes in vivo persistence and enhances anti-tumor activity [J]. Cell Stem Cell, 2020, 27(2): 224-237.e6.
[119] Donnelly R P, Loftus R M, Keating S E, et al. mTORC1-dependent metabolic reprogramming is a prerequisite for NK cell effector function [J]. The Journal of Immunology, 2014, 193(9): 4477-4484.
[120] Rafei H, Basar R, Acharya S, et al. CREM is a regulatory checkpoint of CAR and IL-15 signalling in NK cells [J]. Nature, 2025, 643(8073): 1076-1086.
[121] Wang M J, Krueger J B, Gilkey A K, et al. Precision enhancement of CAR-NK cells through non-viral engineering and highly multiplexed base editing [J]. Journal for ImmunoTherapy of Cancer, 2025, 13(5): e009560.
[122] Christodoulou I, Ho W J, Marple A, et al. Engineering CAR-NK cells to secrete IL-15 sustains their anti-AML functionality but is associated with systemic toxicities [J]. Journal for ImmunoTherapy of Cancer, 2021, 9(12): e003894.
[123] Di Stasi A, Tey S K, Dotti G, et al. Inducible apoptosis as a safety switch for adoptive cell therapy [J]. New England Journal of Medicine, 2011, 365(18): 1673-1683.
[124] Boztug K, Schmidt M, Schwarzer A, et al. Stem-cell gene therapy for the Wiskott-Aldrich syndrome [J]. The New England Journal of Medicine, 2010, 363(20): 1918-1927.
[125] Bonini C, Ferrari G, Verzeletti S, et al. HSV-TK gene transfer into donor lymphocytes for control of allogeneic graft-versus-leukemia [J]. Science, 1997, 276(5319): 1719-1724.
[126] Ciceri F, Bonini C, Stanghellini M T L, et al. Infusion of suicide-gene-engineered donor lymphocytes after family haploidentical haemopoietic stem-cell transplantation for leukaemia (the TK007 trial): A non-randomised phase I–II study [J]. The Lancet Oncology, 2009, 10(5): 489-500.
[127] Sheikh S, Ernst D, Keating A. Prodrugs and prodrug-activated systems in gene therapy [J]. Molecular Therapy, 2021, 29(5): 1716-1728.
[128] Cloughesy T F, Petrecca K, Walbert T, et al. Effect of vocimagene amiretrorepvec in combination with flucytosine vs standard of care on survival following tumor resection in patients with recurrent high-grade glioma: A randomized clinical trial [J]. JAMA Oncology, 2020, 6(12): 1939-1946.
[129] Fan L F, Freeman K W, Khan T, et al. Improved artificial death switches based on caspases and FADD [J]. Human Gene Therapy, 1999, 10(14): 2273-2285.
[130] Li P, Zhou L B, Zhao T, et al. Caspase-9: Structure, mechanisms and clinical application [J]. Oncotarget, 2017, 8(14): 23996-24008.
[131] Lei W, Liu H, Deng W H, 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 [J]. Nature Cancer, 2025, 6(5): 786-800.
[132] Carfagnini C, Singh R, Bechara S B, et al. The efficacy and safety of CD-19 directed CAR-NK therapy in adults with B-cell malignancies: A meta-analysis [J]. Blood, 2024, 144: 7180.
[133] Xu J, Wang B Y, Yu S H, et al. Long-term remission and survival in patients with relapsed or refractory multiple myeloma after treatment with LCAR-B38M CAR T cells: 5-year follow-up of the LEGEND-2 trial [J]. Journal of Hematology & Oncology, 2024, 17(1): 23.
[134] Jagannath S, Martin T G, Lin Y, et al. Long-term (≥5-year) remission and survival after treatment with ciltacabtagene autoleucel in CARTITUDE-1 patients with relapsed/refractory multiple myeloma [J]. Journal of Clinical Oncology, 2025, 43(25): 2766-2771.
[135] Park E, Mun H J, Seo E, et al. CAR NK92 cells targeting BCMA can effectively kill multiple myeloma cells both in vitro and in vivo [J]. Biomedicines, 2024, 12(1): 248.
[136] Motais B, Charvátová S, Walek Z, et al. NK92 expressing anti-BCMA CAR and secreted TRAIL for the treatment of multiple myeloma: Preliminary in vitro assessment [J]. Cells, 2023, 12(23): 2748.
[137] Bras A E, de Haas V, van Stigt A, et al. CD123 expression levels in 846 acute leukemia patients based on standardized immunophenotyping [J]. Cytometry Part B: Clinical Cytometry, 2019, 96(2): 134-142.
[138] Testa U, Pelosi E, Frankel A. CD 123 is a membrane biomarker and a therapeutic target in hematologic malignancies [J]. Biomarker Research, 2014, 2(1): 4.
[139] Caruso S, De Angelis B, Del Bufalo F, et al. Safe and effective off-the-shelf immunotherapy based on CAR.CD123-NK cells for the treatment of acute myeloid leukaemia [J]. Journal of Hematology & Oncology, 2022, 15(1): 163.
[140] Murakami T, Nakazawa T, Natsume A, et al. Novel human NK cell line carrying CAR targeting EGFRvIII induces antitumor effects in glioblastoma cells [J]. Anticancer Research, 2018, 38(9): 5049-5056.
[141] Zhang C C, Burger M C, Jennewein L, et al. ErbB2/HER2-specific NK cells for targeted therapy of glioblastoma [J]. JNCI: Journal of the National Cancer Institute, 2016, 108(5): djv375.
[142] Liu Y, Zhou Y H, Huang K H, et al. Targeting epidermal growth factor-overexpressing triple-negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor [J]. Cell Proliferation, 2020, 53(8): e12858.
[143] Hu Z W. Tissue factor as a new target for CAR-NK cell immunotherapy of triple-negative breast cancer [J]. Scientific Reports, 2020, 10: 2815.
[144] Schönfeld K, Sahm C, Zhang C C, et al. Selective inhibition of tumor growth by clonal NK cells expressing an ErbB2/HER2-specific chimeric antigen receptor [J]. Molecular Therapy, 2015, 23(2): 330-338.
[145] Wu M D, He J, Geng J X, et al. Robo1 CAR-NK92 and radiotherapy exert synergistic efficacy in solid tumors [J]. Journal of Translational Medicine, 2025, 23(1): 720.
[146] Yu M, Luo H, Fan M L, et al. Development of GPC3-specific chimeric antigen receptor-engineered natural killer cells for the treatment of hepatocellular carcinoma [J]. Molecular Therapy, 2018, 26(2): 366-378.
[147] Quintarelli C, Sivori S, Caruso S, et al. Efficacy of third-party chimeric antigen receptor modified peripheral blood natural killer cells for adoptive cell therapy of B-cell precursor acute lymphoblastic leukemia [J]. Leukemia, 2020, 34(4): 1102-1115.
[148] Ng Y Y, Du Z C, Zhang X, et al. CXCR4 and anti-BCMA CAR co-modified natural killer cells suppress multiple myeloma progression in a xenograft mouse model [J]. Cancer Gene Therapy, 2022, 29(5): 475-483.
[149] Ren Q, Zu Y L, Su H C, et al. Single VHH-directed BCMA CAR-NK cells for multiple myeloma [J]. Experimental Hematology & Oncology, 2023, 12(1): 98.
[150] Chen K H, Wada M, Firor A E, et al. Novel anti-CD3 chimeric antigen receptor targeting of aggressive T cell malignancies [J]. Oncotarget, 2016, 7(35): 56219-56232.
[151] Zu Y L, Ren Q, Zhang J S, et al. Targeting CD5 chimeric antigen receptor-engineered natural killer cells against T-cell malignancies [J]. Experimental Hematology & Oncology, 2024, 13(1): 104.
[152] You F T, Wang Y Y, Jiang L C, et al. A novel CD7 chimeric antigen receptor-modified NK-92MI cell line targeting T-cell acute lymphoblastic leukemia [J]. American Journal of Cancer Research, 2019, 9(1): 64-78.
[153] Teng K Y, Mansour A G, Zhu Z, et al. Off-the-shelf prostate stem cell antigen–directed chimeric antigen receptor natural killer cell therapy to treat pancreatic cancer [J]. Gastroenterology, 2022, 162(4): 1319-1333.
[154] Han J F, Chu J H, Keung Chan W, et al. CAR-engineered NK cells targeting wild-type EGFR and EGFRvIII enhance killing of glioblastoma and patient-derived glioblastoma stem cells [J]. Scientific Reports, 2015, 5: 11483.
[155] Müller N, Michen S, Tietze S, et al. Engineering NK cells modified with an EGFRvIII-specific chimeric antigen receptor to overexpress CXCR4 improves immunotherapy of CXCL12/SDF-1α-secreting glioblastoma [J]. Journal of Immunotherapy, 2015, 38(5): 197-210.
[156] Klapdor R, Wang S, Morgan M, et al. Characterization of a novel third-generation anti-CD24-CAR against ovarian cancer [J]. International Journal of Molecular Sciences, 2019, 20(3): 660.
[157] Klapdor R, Wang S, Morgan M A, et al. NK cell-mediated eradication of ovarian cancer cells with a novel chimeric antigen receptor directed against CD44 [J]. Biomedicines, 2021, 9(10): 1339.
[158] Klapdor R, Wang S, Hacker U, et al. Improved killing of ovarian cancer stem cells by combining a novel chimeric antigen receptor–based immunotherapy and chemotherapy [J]. Human Gene Therapy, 2017, 28(10): 886-896.
[159] Dickinson M, Hamad N, Bryant C, et al. s261: First in human data of nkx019, an allogeneic car nk for the treatment of relapsed/refractory (r/r) b-cell malignancies [J]. HemaSphere, 2023, 7(S3): e37234fb.
[160] Bachanova V, Ghobadi A, Patel K, et al. Safety and efficacy of FT596, a first-in-class, multi-antigen targeted, off-the-shelf, iPSC-derived CD19 CAR NK cell therapy in relapsed/refractory B-cell lymphoma [J]. Blood, 2021, 138: 823.
[161] Huang R H, Wen Q, Wang X Q, et al. Off-the-shelf CD33 CAR-NK cell therapy for relapse/refractory AML: First-in-human, phase I trial [J]. Blood, 2022, 140(Supplement 1): 7450-7451.
[162] Tang X, Yang L, Li Z, et al. First-in-man clinical trial of CAR NK-92 cells: Safety test of CD33-CAR NK-92 cells in patients with relapsed and refractory acute myeloid leukemia [J]. American Journal of Cancer Research, 2018, 8(6): 1083-1089.
[163] Li B, Zhu X D, Ge J Y, et al. Intraperitoneal infusion of NKG2D CAR-NK cells induces endogenous CD8+ T cell activation in patients with advanced colorectal cancer [J]. Molecular Therapy, 2025, 33(9): 4509-4528.
[164] Sauter C S, Borthakur G, Mountjoy L, et al. A phase 1 study of NKX101, a chimeric antigen receptor natural killer (CAR-NK) cell therapy, with fludarabine and cytarabine in patients with acute myeloid leukemia [J]. Blood, 2023, 142: 2097.
[165] Xiao L, Cen D Z, Gan H N, et al. Adoptive transfer of NKG2D CAR mRNA-engineered natural killer cells in colorectal cancer patients [J]. Molecular Therapy, 2019, 27(6): 1114-1125.
[166] Seery T E, Nangia C S, Reid P D, et al. Overall survival in patients with metastatic or locally advanced pancreatic cancer following chemoradiation with novel combination of aldoxorubicin, N-803 IL-15 superagonist, and PDL1- NK cell therapy [J]. Journal of Clinical Oncology, 2023, 41(4_suppl): 720.
[167] Seery T E, Kistler M, Lee J H, et al. Quilt-3.064: An open-label phase I study of PD-L1 t-haNK in subjects with locally advanced or metastatic solid cancers [J]. Journal of Clinical Oncology, 2020, 38(15_suppl): TPS3152.
[168] Zhang X D, Guo Y Y, Ji Y H, et al. Cytokine release syndrome after modified CAR-NK therapy in an advanced non-small cell lung cancer patient: A case report [J]. Cell Transplantation, 2022, 31: 09636897221094244.
[169] Dhakal B, Berdeja J G, Gregory T, et al. Interim phase I clinical data of FT576 as monotherapy and in combination with daratumumab in subjects with relapsed/refractory multiple myeloma [J]. Blood, 2022, 140(Supplement 1): 4586-4587.
[170] Shi H, Lin Y N, Ran J, et al. Abstract CT099: First-in-human study of ALF501, polypeptide PSMA-targeted chimeric antigen receptor engineered natural killer cells) for castration-resistant prostate cancer [J]. Cancer Research, 2023, 83(8_Supplement): CT099.
[171] Li C J, Yang N N, Li H S, et al. Robo1-specific chimeric antigen receptor natural killer cell therapy for pancreatic ductal adenocarcinoma with liver metastasis [J]. Journal of Cancer Research and Therapeutics, 2020, 16(2): 393-396.
[172] Li Q, Wang Y, Lin M, et al. Abstract A014: Phase I clinical trial with PD-1/MUC1 CAR-pNK92 immunotherapy [J]. Cancer Immunology Research, 2019, 7(2_Supplement): A014.
[173] Burger M C, Forster M T, Romanski A, et al. Intracranial injection of natural killer cells engineered with a HER2-targeted chimeric antigen receptor in patients with recurrent glioblastoma [J]. Neuro-Oncology, 2023, 25(11): 2058-2071.
[174] Khoshandam M, Soltaninejad H, Mousazadeh M, et al. Clinical applications of the CRISPR/Cas9 genome-editing system: Delivery options and challenges in precision medicine [J]. Genes & Diseases, 2024, 11(1): 268-282.
[175] Ahi Y S, Bangari D S, Mittal S K. Adenoviral vector immunity: Its implications and circumvention strategies [J]. Current Gene Therapy, 2011, 11(4): 307-320.
[176] Lino C A, Harper J C, Carney J P, et al. Delivering CRISPR: A review of the challenges and approaches [J]. Drug Delivery, 2018, 25(1): 1234-1257.
[177] Charlesworth C T, Deshpande P S, Dever D P, et al. Identification of preexisting adaptive immunity to Cas9 proteins in humans [J]. Nature Medicine, 2019, 25(2): 249-254.
[178] Fu Y F, Foden J A, Khayter C, et al. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells [J]. Nature Biotechnology, 2013, 31(9): 822-826.
[179] Doench J G, Fusi N, Sullender M, et al. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9 [J]. Nature Biotechnology, 2016, 34(2): 184-191.
[180] Kleinstiver B P, Pattanayak V, Prew M S, et al. High-fidelity CRISPR–Cas9 nucleases with no detectable genome-wide off-target effects [J]. Nature, 2016, 529(7587): 490-495.
[181] Leibowitz M L, Papathanasiou S, Doerfler P A, et al. Chromothripsis as an on-target consequence of CRISPR–Cas9 genome editing [J]. Nature Genetics, 2021, 53(6): 895-905.
[182] Stadtmauer E A, Fraietta J A, Davis M M, et al. CRISPR-engineered T cells in patients with refractory cancer [J]. Science, 2020, 367(6481): eaba7365.
[183] Cullot G, Boutin J, Toutain J, et al. CRISPR-Cas9 genome editing induces megabase-scale chromosomal truncations [J]. Nature Communications, 2019, 10: 1136.
[184] Kosicki M, Tomberg K, Bradley A. Repair of double-strand breaks induced by CRISPR–Cas9 leads to large deletions and complex rearrangements [J]. Nature Biotechnology, 2018, 36(8): 765-771.
[185] Doss M X, Sachinidis A. Current challenges of iPSC-based disease modeling and therapeutic implications [J]. Cells, 2019, 8(5): 403.
[186] Yoshihara M, Hayashizaki Y, Murakawa Y. Genomic instability of iPSCs: Challenges towards their clinical applications [J]. Stem Cell Reviews and Reports, 2017, 13(1): 7-16.
[187] Buckberry S, Liu X D, Poppe D, et al. Transient naive reprogramming corrects hiPS cells functionally and epigenetically [J]. Nature, 2023, 620(7975): 863-872.
[188] Gore A, Li Z, Fung H L, et al. Somatic coding mutations in human induced pluripotent stem cells [J]. Nature, 2011, 471(7336): 63-67.
[189] Dashtban M, Panchalingam K M, Shafa M, et al. Addressing manufacturing challenges for commercialization of iPSC-based therapies [M]//Stem cells and good manufacturing practices. New York: Springer, 2020: 179-198.
[190] Hermanson D L, Ni Z Y, Kaufman D S. Human pluripotent stem cells as a renewable source of natural killer cells [M]//Hematopoietic differentiation of human pluripotent stem cells. Dordrecht: Springer, 2015: 69-79.
[191] Zhu H, Kaufman D S. An improved method to produce clinical-scale natural killer cells from human pluripotent stem cells [M]//In vitro differentiation of T-cells. New York: Springer, 2019: 107-119.
[192] Koehl U, Kalberer C, Spanholtz J, et al. Advances in clinical NK cell studies: Donor selection, manufacturing and quality control [J]. OncoImmunology, 2016, 5(4): e1115178.
[193] Laskowski T J, Biederstädt A, Rezvani K. Natural killer cells in antitumour adoptive cell immunotherapy [J]. Nature Reviews Cancer, 2022, 22(10): 557-575.
[194] Miller J S, Soignier Y, Panoskaltsis-Mortari A, et al. Successful adoptive transfer and in vivo expansion of human haploidentical NK cells in patients with cancer [J]. Blood, 2005, 105(8): 3051-3057.
[195] Agarwal S, Weidner T, Thalheimer F B, et al. In vivo generated human CAR T cells eradicate tumor cells [J]. OncoImmunology, 2019, 8(12): e1671761.
[196] Xu J, Liu L, Parone P, et al. In-vivo B-cell maturation antigen CAR T-cell therapy for relapsed or refractory multiple myeloma [J]. The Lancet, 2025, 406(10500): 228-231.
[197] Bot A, Scharenberg A, Friedman K, et al. In vivo chimeric antigen receptor (CAR)-T cell therapy [J]. Nature Reviews Drug Discovery, 2026, 25(2): 116-137.
[198] Garrison B, Deng H, Yucel G, et al. Senti-202, a selective, off-the-shelf, preclinical CAR-NK cell therapy with CD33 and/or FLT3 activating CAR, healthy cell protection from endomucin (EMCN) inhibitory CAR and calibrated release IL-15 for hematologic malignancies including AML [J]. Blood, 2022, 140(Supplement 1): 4531-4532.