Goethe University Frankfurt, Department of Pediatrics, Experimental Immunology & Cell Therapy, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany.
Frankfurt Cancer Institute, Goethe University, Paul-Ehrlich-Straße 42-44, 60596 Frankfurt am Main, Germany.
J Leukoc Biol. 2023 Oct 26;114(5):475-486. doi: 10.1093/jleuko/qiad074.
Natural killer cells are a promising platform for cancer immunotherapy. Natural killer cells have high intrinsic killing capability, and the insertion of a chimeric antigen receptor can further enhance their antitumor potential. In first-in-human trials, chimeric antigen receptor-natural killer cells demonstrated strong clinical activity without therapy-induced side effects. The applicability of natural killer cells as an "off-the-shelf" product makes them highly attractive for gene-engineered cell therapies. Traditionally, viral transduction has been used for gene editing; however, the use of viral vectors remains a safety concern and is associated with high costs and regulatory requirements. Here, we review the current landscape of nonviral approaches for chimeric antigen receptor-natural killer cell generation. This includes transfection of vector particles and electroporation of mRNA and DNA vectors, resulting in transient modification and chimeric antigen receptor expression. In addition, using nonviral transposon technologies, natural killer cells can be stably modified ensuring long-lasting chimeric antigen receptor expression. Finally, we discuss CRISPR/Cas9 tools to edit key genes for natural killer cell functionality.
自然杀伤细胞是癌症免疫疗法的一个有前途的平台。自然杀伤细胞具有很高的内在杀伤能力，而嵌合抗原受体的插入可以进一步增强其抗肿瘤潜力。在首次人体试验中，嵌合抗原受体自然杀伤细胞表现出强烈的临床活性，没有治疗诱导的副作用。自然杀伤细胞作为“现成”产品的适用性使它们成为基因工程细胞疗法的极具吸引力。传统上，病毒转导已用于基因编辑；然而，病毒载体的使用仍然存在安全隐患，并且与高成本和监管要求相关。在这里，我们回顾了用于嵌合抗原受体自然杀伤细胞生成的非病毒方法的现状。这包括载体颗粒的转染和 mRNA 和 DNA 载体的电穿孔，导致瞬时修饰和嵌合抗原受体表达。此外，使用非病毒转座子技术，自然杀伤细胞可以进行稳定修饰，确保嵌合抗原受体的长期表达。最后，我们讨论了用于编辑自然杀伤细胞功能的关键基因的 CRISPR/Cas9 工具。
