T Cell Therapeutics Research Labs, Cellular Immunotherapy Center, Department of Hematology and Hematopoietic Cell Transplantation, City of Hope, Duarte, California.
Division of Regenerative Medicine, Department of Medicine, University of California, San Diego, San Diego, California.
Cancer Discov. 2021 May;11(5):1192-1211. doi: 10.1158/2159-8290.CD-20-1243. Epub 2020 Dec 16.
Glioblastoma (GBM) contains self-renewing GBM stem cells (GSC) potentially amenable to immunologic targeting, but chimeric antigen receptor (CAR) T-cell therapy has demonstrated limited clinical responses in GBM. Here, we interrogated molecular determinants of CAR-mediated GBM killing through whole-genome CRISPR screens in both CAR T cells and patient-derived GSCs. Screening of CAR T cells identified dependencies for effector functions, including TLE4 and IKZF2. Targeted knockout of these genes enhanced CAR antitumor efficacy. Bulk and single-cell RNA sequencing of edited CAR T cells revealed transcriptional profiles of superior effector function and inhibited exhaustion responses. Reciprocal screening of GSCs identified genes essential for susceptibility to CAR-mediated killing, including RELA and NPLOC4, the knockout of which altered tumor-immune signaling and increased responsiveness of CAR therapy. Overall, CRISPR screening of CAR T cells and GSCs discovered avenues for enhancing CAR therapeutic efficacy against GBM, with the potential to be extended to other solid tumors. SIGNIFICANCE: Reciprocal CRISPR screening identified genes in both CAR T cells and tumor cells regulating the potency of CAR T-cell cytotoxicity, informing molecular targeting strategies to potentiate CAR T-cell antitumor efficacy and elucidate genetic modifications of tumor cells in combination with CAR T cells to advance immuno-oncotherapy..
胶质母细胞瘤（GBM）包含自我更新的胶质母细胞瘤干细胞（GSC），可能适合免疫靶向治疗，但嵌合抗原受体（CAR）T 细胞治疗在 GBM 中的临床反应有限。在这里，我们通过 CAR T 细胞和患者来源的 GSCs 中的全基因组 CRISPR 筛选来研究 CAR 介导的 GBM 杀伤的分子决定因素。CAR T 细胞的筛选确定了效应功能的依赖性，包括 TLE4 和 IKZF2。这些基因的靶向敲除增强了 CAR 的抗肿瘤疗效。编辑后的 CAR T 细胞的批量和单细胞 RNA 测序揭示了具有优越效应功能和抑制耗竭反应的转录谱。GSCs 的相互筛选确定了对 CAR 介导杀伤敏感的必需基因，包括 RELA 和 NPLOC4，敲除这些基因改变了肿瘤免疫信号，并增加了 CAR 治疗的反应性。总体而言，CAR T 细胞和 GSCs 的 CRISPR 筛选发现了增强 CAR 治疗 GBM 疗效的途径，并有潜力扩展到其他实体瘤。意义：相互 CRISPR 筛选鉴定了 CAR T 细胞和肿瘤细胞中的基因，这些基因调节 CAR T 细胞细胞毒性的效力，为增强 CAR T 细胞抗肿瘤疗效提供了分子靶向策略，并阐明了与 CAR T 细胞联合使用的肿瘤细胞的遗传修饰，以推进免疫肿瘤治疗。
