Chimeric antigen receptor (CAR)-natural killer (NK) cell therapies hold promise for solid tumours but remain limited because of poor tumour infiltration, persistence and resistance in the tumour microenvironment1,2,3,4. Here, to identify gain-of-function targets that enhance CAR-NK cell efficacy, we performed an unbiased in vivo CRISPR activation screen followed by a barcoded targeted in vivo open reading frame screen in primary human CAR-NK cells. We identified and comprehensively validated OR7A10, a G protein-coupled receptor (GPCR), as the top candidate. Engineering CAR-NK cells with OR7A10 cDNA (a CRISPR-independent method with a simple manufacturing strategy) enhanced their proliferation, activation, degranulation, cytokine production, death ligand expression, chemokine receptor expression, cytotoxicity, persistence, metabolic fitness and tumour microenvironment resistance. Moreover, exhaustion in primary human NK cells derived from multiple peripheral blood and cord blood donors was reduced. OR7A10 gain-of-function CAR-NK cells displayed strong in vivo efficacy across multiple solid tumour models. For example, 100% complete response with long-term tumour control and survival benefit in an orthotopic breast cancer mouse model were achieved. These findings establish OR7A10-engineered CAR-NK cells as a highly potent and scalable off-the-shelf therapeutic for solid tumours.
嵌合抗原受体（CAR）-自然杀伤（NK）细胞疗法为实体瘤治疗带来希望，但由于肿瘤浸润不足、持久性差以及肿瘤微环境中的抵抗性，其应用仍受限[1-4]。为筛选能增强CAR-NK细胞效能的功能获得性靶点，我们首先在原代人CAR-NK细胞中开展了无偏倚的体内CRISPR激活筛选，随后进行了条形码靶向性体内开放阅读框筛选。通过鉴定与全面验证，我们发现G蛋白偶联受体（GPCR）OR7A10为最优候选靶点。采用OR7A10 cDNA（一种无需CRISPR、制备策略简单的工程化方法）改造的CAR-NK细胞，其增殖能力、活化水平、脱颗粒功能、细胞因子分泌、死亡配体表达、趋化因子受体表达、细胞毒性、持久性、代谢适应度及肿瘤微环境抵抗性均显著提升。此外，来源于多名外周血和脐带血供体的原代人NK细胞耗竭现象亦得到改善。在多种实体瘤模型中，OR7A10功能获得性CAR-NK细胞展现出强大的体内疗效——例如在原位乳腺癌小鼠模型中实现了100%完全缓解，并带来长期肿瘤控制与生存获益。本研究确立OR7A10工程化CAR-NK细胞作为治疗实体瘤的高效、可扩展的现货型细胞疗法。
OR7A10 GPCR engineering boosts CAR-NK therapy against solid tumours
肿瘤(癌症)患者之家