Genome-wide CRISPR/Cas9 screen reveals factors that influence the susceptibility of tumor cells to NK cell-mediated killing.

IF 10.3 1区 医学 Q1 IMMUNOLOGY
Sophie Guia, Aurore Fenis, Camille Baudesson De Chanville, Justine Galluso, Hakim Medjouel, Bertrand Escaliere, Angelika Modelska, Margaux Vienne, Noella Lopes, Amelie Pouchin, Benjamin Rossi, Laurent Gauthier, Sandrine Roulland, Eric Vivier, Emilie Narni-Mancinelli
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Abstract

Background: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

全基因组CRISPR/Cas9筛选揭示了影响肿瘤细胞对NK细胞介导的杀伤易感性的因素。
背景:自然杀伤(NK)细胞对各种类型的癌细胞表现出强大的细胞毒活性。在过去的五十年中,许多方法被用来阐明NK细胞介导的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NK细胞介导的肿瘤细胞破坏的调节因子尚未完全了解。这包括NK细胞识别的多种肿瘤配体以及NK细胞用来消除肿瘤细胞的机制。方法:在本研究中,我们采用全基因组CRISPR/Cas9筛选方法,结合功能性细胞毒性试验,描绘了调节结肠癌HCT-116细胞对NK细胞介导的细胞毒性易感性的途径。结果:对与NK细胞共孵生后存活的HCT-116细胞中引导RNA分布的分析发现,ICAM-1在NKp44介导的免疫突触中起关键作用,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信号通路的基因的破坏赋予了对NK细胞攻击的抵抗力。我们进一步分析了NK细胞来源的IFN-γ在体外促进线粒体凋亡,并在体内控制B16-F10肺转移。结论:在NK细胞为基础的肿瘤治疗中,监测肿瘤细胞表面的ICAM-1水平或调节其表达是值得考虑的。此外,促进NK细胞表面FasL的表达被重申为增强NK细胞介导的肿瘤杀伤的重要策略,为优化治疗提供了额外的途径。此外,考虑到IFN-γ的扩散特性,我们的研究结果强调了利用NK细胞来源的IFN-γ增强直接杀伤肿瘤细胞和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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal for Immunotherapy of Cancer
Journal for Immunotherapy of Cancer Biochemistry, Genetics and Molecular Biology-Molecular Medicine
CiteScore
17.70
自引率
4.60%
发文量
522
审稿时长
18 weeks
期刊介绍: The Journal for ImmunoTherapy of Cancer (JITC) is a peer-reviewed publication that promotes scientific exchange and deepens knowledge in the constantly evolving fields of tumor immunology and cancer immunotherapy. With an open access format, JITC encourages widespread access to its findings. The journal covers a wide range of topics, spanning from basic science to translational and clinical research. Key areas of interest include tumor-host interactions, the intricate tumor microenvironment, animal models, the identification of predictive and prognostic immune biomarkers, groundbreaking pharmaceutical and cellular therapies, innovative vaccines, combination immune-based treatments, and the study of immune-related toxicity.
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