Systematic Immunotherapy Target Discovery Using Genome-Scale In Vivo CRISPR Screens in CD8 T Cells
Systematic Immunotherapy Target Discovery Using Genome-Scale In Vivo CRISPR Screens in CD8 T Cells
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DOI：
10.1016/j.cell.2019.07.044
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发表时间：
2019-08-22
期刊：
影响因子：
64.5
通讯作者：
Chen, Sidi
中科院分区：
文献类型：
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作者：
Dong, Matthew B.;Wang, Guangchuan;Chen, Sidi
CD 8 T细胞在抗肿瘤免疫应答中发挥重要作用。在这里，我们直接在癌症免疫治疗环境下在CD 8 T细胞中进行了基因组规模的CRISPR筛选，并确定了肿瘤浸润和脱粒的调节因子。体内筛选有力地重新鉴定了典型的免疫治疗靶标，例如PD-1和Tim-3，沿着有尚未在T细胞中表征的基因。浸润和脱粒筛选集中在RNA解旋酶Dhx 37上。Dhx 37基因敲除增强了抗原特异性CD 8 T细胞在体内对抗三阴性乳腺癌的功效。小鼠和人CD 8 T细胞中的免疫学表征揭示了DHX 37抑制效应子功能、细胞因子产生和T细胞活化。转录组学分析和生化分析揭示了DHX 37在调节NF-κ B中的作用。这些数据证明了用于免疫治疗靶标发现的高通量体内遗传筛选，并确立了DHX 37作为CD 8 T细胞的功能调节剂。
CD8 T cells play essential roles in anti-tumor immune responses. Here, we performed genome-scale CRISPR screens in CD8 T cells directly under cancer immunotherapy settings and identified regulators of tumor infiltration and degranulation. The in vivo screen robustly re-identified canonical immunotherapy targets such as PD-1 and Tim-3, along with genes that have not been characterized in T cells. The infiltration and degranulation screens converged on an RNA helicase Dhx37. Dhx37 knockout enhanced the efficacy of antigen-specific CD8 T cells against triple-negative breast cancer in vivo. Immunological characterization in mouse and human CD8 T cells revealed that DHX37 suppresses effector functions, cytokine production, and T cell activation. Transcriptomic profiling and biochemical interrogation revealed a role for DHX37 in modulating NF-kappa B. These data demonstrate high-throughput in vivo genetic screens for immunotherapy target discovery and establishes DHX37 as a functional regulator of CD8 T cells.