文章：

用于实体瘤免疫检查点药物筛选的GAS-Luc2报告基因细胞系

GAS-Luc2 Reporter Cell Lines for Immune Checkpoint Drug Screening in Solid Tumors

原文发布日期：22 May 2024

DOI: 10.3390/cancers16111965

类型: Article

开放获取: 是

英文摘要：

Recent studies highlight the integral role of the interferon gamma receptor (IFNγR) pathway in T cell–mediated cytotoxicity against solid but not liquid tumors. IFNγ not only directly facilitates tumor cell death by T cells but also indirectly promotes cytotoxicity via myeloid phagocytosis in the tumor microenvironment. Meanwhile, full human ex vivo immune checkpoint drug screening remains challenging. We hypothesized that an engineered gamma interferon activation site response element luciferase reporter (GAS-Luc2) can be utilized for immune checkpoint drug screening in diverse ex vivo T cell–solid tumor cell co-culture systems. We comprehensively profiled cell surface proteins in ATCC’s extensive collection of human tumor and immune cell lines, identifying those with endogenously high expression of established and novel immune checkpoint molecules and binding ligands. We then engineered three GAS-Luc2 reporter tumor cell lines expressing immune checkpoints PD-L1, CD155, or B7-H3/CD276. Luciferase expression was suppressed upon relevant immune checkpoint–ligand engagement. In the presence of an immune checkpoint inhibitor, T cells released IFNγ, activating the JAK-STAT pathway in GAS-Luc2 cells, and generating a quantifiable bioluminescent signal for inhibitor evaluation. These reporter lines also detected paracrine IFNγ signaling for immune checkpoint-targeted ADCC drug screening. Further development into an artificial antigen-presenting cell line (aAPC) significantly enhanced T cell signaling for superior performance in these ex vivo immune checkpoint drug screening platforms.

摘要翻译： 

近期研究强调了γ干扰素受体（IFNγR）通路在T细胞介导的实体瘤（而非液体肿瘤）细胞毒性中的关键作用。IFNγ不仅直接促进T细胞杀伤肿瘤细胞，还能通过肿瘤微环境中的髓系细胞吞噬作用间接增强细胞毒性。然而，全人源离体免疫检查点药物筛选仍面临挑战。我们提出假设：工程化的γ干扰素激活位点反应元件荧光素酶报告系统（GAS-Luc2）可用于多种离体T细胞-实体瘤细胞共培养体系的免疫检查点药物筛选。通过对ATCC大规模人源肿瘤及免疫细胞系库的细胞表面蛋白进行系统性表征，我们筛选出内源性高表达经典及新型免疫检查点分子及其配体的细胞系。随后构建了三种表达免疫检查点PD-L1、CD155或B7-H3/CD276的GAS-Luc2报告肿瘤细胞系。当相应免疫检查点与配体结合时，荧光素酶表达受到抑制；而在免疫检查点抑制剂存在下，T细胞释放的IFNγ通过激活GAS-Luc2细胞中的JAK-STAT通路，产生可量化的生物发光信号用于抑制剂评估。这些报告细胞系还能检测旁分泌IFNγ信号，适用于靶向免疫检查点的抗体依赖性细胞介导的细胞毒性（ADCC）药物筛选。进一步开发的人工抗原呈递细胞系（aAPC）显著增强了T细胞信号传导，使该离体免疫检查点药物筛选平台性能得到全面提升。

原文链接：

GAS-Luc2 Reporter Cell Lines for Immune Checkpoint Drug Screening in Solid Tumors