OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium.
Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium.
Front Immunol. 2024 Oct 21;15:1444886. doi: 10.3389/fimmu.2024.1444886. eCollection 2024.
Cancer cells effectively evade immune surveillance, not only through the well-known PD-1/PD-L1 pathway but also alternative mechanisms that impair patient response to immune checkpoint inhibitors. We present a novel co-culture model that pairs a reporter T-cell line with different melanoma cell lines that have varying immune evasion characteristics. We developed a scalable high-throughput lentiviral arrayed CRISPR interference (CRISPRi) screening protocol to conduct gene perturbations in both T-cells and melanoma cells, enabling the identification of genes that modulate tumor immune evasion. Our study functionally validates the co-culture model system and demonstrates the performance of the CRISPRi-screening protocol by modulating the expression of known regulators of tumor immunity. Together, our work provides a robust framework for future research aimed at systematically exploring mechanisms of tumor immune evasion.
癌细胞有效地逃避了免疫监视，不仅通过众所周知的 PD-1/PD-L1 途径，而且还通过损害患者对免疫检查点抑制剂反应的其他替代机制。我们提出了一种新的共培养模型，该模型将报告 T 细胞系与具有不同免疫逃逸特征的不同黑色素瘤细胞系配对。我们开发了一种可扩展的高通量慢病毒阵列 CRISPR 干扰 (CRISPRi) 筛选方案，对 T 细胞和黑色素瘤细胞进行基因扰动，从而确定调节肿瘤免疫逃逸的基因。我们的研究从功能上验证了共培养模型系统，并通过调节肿瘤免疫已知调节剂的表达来展示 CRISPRi 筛选方案的性能。总之，我们的工作为旨在系统探索肿瘤免疫逃逸机制的未来研究提供了一个强大的框架。
