自然杀伤细胞可以在没有事先敏感的情况下攻击癌细胞，但由于缺乏特异性受体以靶向肿瘤细胞，其临床效益有限。在本研究中，我们旨在使NK细胞能够以特异性靶向胞内甘油脂受体3+的肿瘤细胞，同时不产生细胞损伤或遗传改变，并进一步评估其治疗效果。采用代谢糖修饰法通过细胞表面的Gpc3 DNA适配体对NK细胞进行修饰，赋予其特异性靶向能力。然后，在体外评估G-NK细胞的特异性靶向特性、细胞毒性和细胞因子分泌情况。最后，在体内研究了G-NK细胞对甘油脂受体3+肿瘤细胞的治疗效果。与随机适配体突变修饰的NK细胞和未修饰的NK细胞相比，G-NK细胞诱导GPC3+肿瘤细胞显著凋亡/坏死，并分泌细胞因子以维持强烈的细胞毒活性。此外，由于G-NK细胞在肿瘤部位的富集增强，G-NK细胞显著抑制了HepG2肿瘤携带小鼠的肿瘤生长。所提出的策略赋予了NK细胞肿瘤特异性靶向能力，提高了对GPC3+肝细胞癌的养护治疗效率。© 2023. Springer Science+Business Media, LLC.

Natural killer cells can attack cancer cells without prior sensitization, but their clinical benefit is limited owing to their poor selectivity that is caused by the lack of specific receptors to target tumor cells. In this study, we aimed to endow NK cells with the ability to specifically target glypican-3+ tumor cells without producing cell damage or genetic alterations, and further evaluated their therapeutic efficiency.NK cells were modified with a Gpc3 DNA aptamer on the cell surface via metabolic glycoengineering to endow NK cells with specific targeting ability. Then, the G-NK cells were evaluated for their specific targeting properties, cytotoxicity and secretion of cytokines in vitro. Finally, we investigated the therapeutic efficiency of G-NK cells against glypican-3+ tumor cells in vivo.Compared with NK cells modified with a random aptamer mutation and unmodified NK cells, G-NK cells induced significant apoptosis/necrosis of GPC3+ tumor cells and secreted cytokines to preserve the intense cytotoxic activities. Moreover, G-NK cells significantly suppressed tumor growth in HepG2 tumor-bearing mice due to the enhanced enrichment of G-NK cells at the tumor site.The proposed strategy endows NK cells with a tumor-specific targeting ability to enhance adoptive therapeutic efficiency in GPC3+ hepatocellular carcinoma.© 2023. Springer Science+Business Media, LLC.