Integrated drug profiling and CRISPR screening identify essential pathways for CAR T-cell cytotoxicity
Integrated drug profiling and CRISPR screening identify essential pathways for CAR T-cell cytotoxicity
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DOI：
10.1182/blood.2019002121
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发表时间：
2020-02-27
期刊：
影响因子：
20.3
通讯作者：
Mustjoki, Satu
中科院分区：
文献类型：
--
作者：
Dufva, Olli;Koski, Jan;Mustjoki, Satu
嵌合抗原受体（CAR）T细胞疗法已被证明对复发性和难治性B细胞恶性肿瘤有效，但仍会发生耐药性和复发。更好地理解影响CAR T细胞细胞毒性的机制以及使用小分子药物进行调节的潜力可以改善当前的免疫疗法。在这里，我们使用超过500种小分子药物和基因组规模的CRISPR-Cas9功能丧失筛选系统地研究了CAR T细胞细胞毒性的药物机制。我们鉴定了几种酪氨酸激酶抑制剂，其通过损害T细胞信号传导转录活性来抑制CAR T细胞的细胞毒性。相比之下，凋亡调节剂药物SMAC模拟物使B细胞急性淋巴细胞白血病和弥漫性大B细胞淋巴瘤细胞对抗CD 19 CAR T细胞敏感。CRISPR筛选鉴定了通过FADD和TNFRSF 10 B（TRAIL-R2）的死亡受体信号传导作为CAR T细胞细胞毒性的关键介体，并阐明了SMAC模拟物的RIPK 1依赖性致敏机制。死亡受体表达在B细胞恶性肿瘤的遗传亚型中存在差异，表明CAR T细胞细胞毒性机制与癌症遗传学之间存在联系。这些结果暗示死亡受体信号传导是癌细胞对CAR T细胞毒性敏感性的重要介质，具有增强癌症免疫治疗的药理学靶向潜力。筛选数据提供了癌症药物的免疫调节特性和影响CAR T细胞细胞毒性的遗传机制的资源。
Chimeric antigen receptor (CAR) T-cell therapy has proven effective in relapsed and refractory B-cell malignancies, but resistance and relapses still occur. Better understanding of mechanisms influencing CAR T-cell cytotoxicity and the potential for modulation using small-molecule drugs could improve current immunotherapies. Here, we systematically investigated druggable mechanisms of CAR T-cell cytotoxicity using >500 small-molecule drugs and genome-scale CRISPR-Cas9 loss-of-function screens. We identified several tyrosine kinase inhibitors that inhibit CAR T-cell cytotoxicity by impairing T-cell signaling transcriptional activity. In contrast, the apoptotic modulator drugs SMAC mimetics sensitized B-cell acute lymphoblastic leukemia and diffuse large B-cell lymphoma cells to anti-CD19 CAR T cells. CRISPR screens identified death receptor signaling through FADD and TNFRSF10B (TRAIL-R2) as a key mediator of CAR T-cell cytotoxicity and elucidated the RIPK1-dependent mechanism of sensitization by SMAC mimetics. Death receptor expression varied across genetic subtypes of B-cellmalignancies, suggesting a link between mechanisms of CAR T-cell cytotoxicity and cancer genetics. These results implicate death receptor signaling as an important mediator of cancer cell sensitivity to CAR T-cell cytotoxicity, with potential for pharmacological targeting to enhance cancer immunotherapy. The screening data provide a resource of immunomodulatory properties of cancer drugs and genetic mechanisms influencing CAR T-cell cytotoxicity.