Childrens' Hospital, Experimental Immunology, Johann Wolfgang Goethe University, Frankfurt am Main, Germany.
Frankfurt Cancer Institute, Goethe University, Frankfurt am Main, Germany.
Oncoimmunology. 2022 May 31;11(1):2081415. doi: 10.1080/2162402X.2022.2081415. eCollection 2022.
Natural Killer (NK) cells are known for their high intrinsic cytotoxic capacity, and the possibility to be applied as 'off-the-shelf' product makes them highly attractive for cell-based immunotherapies. In patients with multiple myeloma (MM), an elevated number of NK cells has been correlated with higher overall-survival rate. However, NK cell function can be impaired by upregulation of inhibitory receptors, such as the immune checkpoint NKG2A. Here, we developed a CRISPR-Cas9-based gene editing protocol that allowed us to knockout about 80% of the NKG2A-encoding () locus in primary NK cells. In-depth phenotypic analysis confirmed significant reduction in NKG2A protein expression. Importantly, the edited NK cells showed significantly increased cytotoxicity against primary MM cells isolated from a small cohort of patients, and maintained the NK cell-specific cytokine production. In conclusion, -editing in primary NK cells has the prospect of overcoming immune checkpoint inhibition in clinical applications.
自然杀伤 (NK) 细胞以其固有高细胞毒性而闻名，并且作为“现成”产品的可能性使它们在细胞免疫疗法中极具吸引力。在多发性骨髓瘤 (MM) 患者中，NK 细胞数量升高与总生存率更高相关。然而，NK 细胞的功能可通过抑制性受体（如免疫检查点 NKG2A）的上调而受损。在这里，我们开发了一种基于 CRISPR-Cas9 的基因编辑方案，该方案使我们能够敲除原代 NK 细胞中约 80%的 NKG2A 编码 () 基因座。深入的表型分析证实了 NKG2A 蛋白表达的显著降低。重要的是，编辑后的 NK 细胞对从小部分患者中分离出的原代 MM 细胞显示出显著增强的细胞毒性，并且保持了 NK 细胞特异性细胞因子的产生。总之，在原代 NK 细胞中进行 - 编辑具有克服临床应用中免疫检查点抑制的前景。
