Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, United States.
Bioinformatics and Computational Biosciences Branch, Office of Cyber Infrastructure and Computational Biology, National Institute of Allergy and Infectious Diseases, U.S. National Institutes of Health, Rockville, MD, United States.
Front Immunol. 2019 Dec 11;10:2879. doi: 10.3389/fimmu.2019.02879. eCollection 2019.
The anti-leukemia activity of NK cells helps prevent relapse during hematopoietic stem cell transplantation (HSCT) in leukemia patients. However, the factors that determine the sensitivity or resistance of leukemia cells in the context of NK-mediated cytotoxicity are not well-established. Here, we performed a genome-wide CRISPR screen in the human chronic-myelogenous-leukemia (CML) cell line K562 to identify genes that regulate the vulnerability of leukemia cells to killing by primary human NK cells. The distribution of guide RNAs (gRNAs) in K562 cells that survived co-incubation with NK cells showed that loss of , which encodes the ligand of the natural cytotoxicity receptor NKp30, protected K562 cells from killing. In contrast, loss of genes that regulate the antigen-presentation and interferon-γ-signaling pathways increased the vulnerability of K562 cells. The addition of IFN-γ neutralizing antibody increased the susceptibility of K562 cells to NK-mediated killing. Upregulation of MHC class I on K562 cells after co-incubation with NK cells was dependent on . Analysis of RNA-seq data from The Cancer Genome Atlas (TCGA) showed that low expression in cancer tissues was associated with improved overall survival in acute myeloid leukemia (AML) and Kidney Renal Clear Cell Carcinoma (KIRC) patients. Our results, showing that the upregulation of MHC class I by NK-derived IFN-γ leads to resistance to NK cytotoxicity, suggest that targeting IFN-γ responses might be a promising approach to enhance NK cell anti-cancer efficacy.
自然杀伤 (NK) 细胞的抗白血病活性有助于预防白血病患者造血干细胞移植 (HSCT) 后的复发。然而，NK 介导的细胞毒性作用下白血病细胞的敏感性或耐药性的决定因素尚未得到充分确立。在这里，我们在人慢性髓系白血病 (CML) 细胞系 K562 中进行了全基因组 CRISPR 筛选，以鉴定调节白血病细胞对原代人 NK 细胞杀伤敏感性的基因。与 NK 细胞共孵育后存活的 K562 细胞中的向导 RNA (gRNA) 的分布表明，缺失编码自然细胞毒性受体 NKp30 配体的 ，可保护 K562 细胞免受杀伤。相比之下，调节抗原呈递和干扰素-γ 信号通路的基因的缺失增加了 K562 细胞的易感性。添加 IFN-γ 中和抗体增加了 K562 细胞对 NK 介导的杀伤的敏感性。与 NK 细胞共孵育后 K562 细胞 MHC Ⅰ类分子的上调依赖于 。对来自癌症基因组图谱 (TCGA) 的 RNA-seq 数据的分析表明，癌症组织中低表达与急性髓系白血病 (AML) 和肾透明细胞癌 (KIRC) 患者的总体生存率提高相关。我们的结果表明，NK 衍生的 IFN-γ 上调 MHC Ⅰ类分子导致对 NK 细胞毒性的耐药性，表明靶向 IFN-γ 反应可能是增强 NK 细胞抗癌疗效的一种有前途的方法。
