Guia Sophie, Fenis Aurore, Baudesson De Chanville Camille, Galluso Justine, Medjouel Hakim, Escaliere Bertrand, Modelska Angelika, Vienne Margaux, Lopes Noella, Pouchin Amelie, Rossi Benjamin, Gauthier Laurent, Roulland Sandrine, Vivier Eric, Narni-Mancinelli Emilie
CIML, Marseille, France.
Aix-Marseille-University, Marseille, France.
J Immunother Cancer. 2025 Mar 31;13(3):e010699. doi: 10.1136/jitc-2024-010699.
Natural killer (NK) cells exhibit potent cytotoxic activity against various cancer cell types. Over the past five decades, numerous methodologies have been employed to elucidate the intricate molecular mechanisms underlying NK cell-mediated tumor control. While significant progress has been made in elucidating the interactions between NK cells and tumor cells, the regulatory factors governing NK cell-mediated tumor cell destruction are not yet fully understood. This includes the diverse array of tumor ligands recognized by NK cells and the mechanisms that NK cells employ to eliminate tumor cells.
In this study, we employed a genome-wide CRISPR/Cas9 screening approach in conjunction with functional cytotoxicity assays to delineate the pathways modulating the susceptibility of colon adenocarcinoma HCT-116 cells to NK cell-mediated cytotoxicity.
Analysis of guide RNA distribution in HCT-116 cells that survived co-incubation with NK cells identified ICAM-1 as a pivotal player in the NKp44-mediated immune synapse, with NKp44 serving as an activating receptor crucial for the elimination of HCT-116 tumor cells by NK cells. Furthermore, disruption of genes involved in the apoptosis or interferon (IFN)-γ signaling pathways conferred resistance to NK cell attack. We further dissected that NK cell-derived IFN-γ promotes mitochondrial apoptosis in vitro and exerts control over B16-F10 lung metastases in vivo.
Monitoring ICAM-1 levels on the surface of tumor cells or modulating its expression should be considered in the context of NK cell-based therapy. Furthermore, promoting FasL expression on the NK cell surface is reaffirmed as an important strategy to enhance NK cell-mediated tumor killing, offering an additional avenue for therapeutic optimization. Additionally, considering the diffusion properties of IFN-γ, our findings highlight the potential of leveraging NK cell-derived IFN-γ to enhance direct tumor cell killing and facilitate bystander effects via cytokine diffusion, warranting further investigation.
自然杀伤（NK）细胞对多种癌细胞类型具有强大的细胞毒性活性。在过去的五十年里，人们采用了众多方法来阐明NK细胞介导肿瘤控制的复杂分子机制。虽然在阐明NK细胞与肿瘤细胞之间的相互作用方面取得了重大进展，但调控NK细胞介导的肿瘤细胞破坏的因素尚未完全了解。这包括NK细胞识别的多种肿瘤配体以及NK细胞用于消除肿瘤细胞的机制。
在本研究中，我们采用全基因组CRISPR/Cas9筛选方法并结合功能细胞毒性测定，以描绘调节结肠腺癌HCT-116细胞对NK细胞介导的细胞毒性敏感性的途径。
对与NK细胞共孵育后存活的HCT-116细胞中引导RNA分布的分析确定细胞间黏附分子-1（ICAM-1）是NKp44介导的免疫突触中的关键参与者，NKp44作为激活受体，对NK细胞消除HCT-116肿瘤细胞至关重要。此外，参与凋亡或干扰素（IFN）-γ信号通路的基因破坏赋予了对NK细胞攻击的抗性。我们进一步剖析发现，NK细胞衍生的IFN-γ在体外促进线粒体凋亡，并在体内对B16-F10肺转移发挥控制作用。
在基于NK细胞的治疗背景下，应考虑监测肿瘤细胞表面的ICAM-1水平或调节其表达。此外，再次确认促进NK细胞表面FasL表达是增强NK细胞介导的肿瘤杀伤的重要策略,为治疗优化提供了另一条途径。此外，考虑到IFN-γ的扩散特性，我们的研究结果突出了利用NK细胞衍生的IFN-γ增强直接肿瘤细胞杀伤并通过细胞因子扩散促进旁观者效应的潜力，值得进一步研究。
