Purpose: Cytokine-engineering of chimeric antigen receptor-redirected T cells (CAR T cells) is a promising principle to overcome the limited activity of canonical CAR T cells against solid cancers. Experimental Design: We developed an investigational medicinal product, GD2IL18CART, consisting of CAR T cells directed against ganglioside GD2 with CAR-inducible IL18 to enhance their activation response and cytolytic effector functions in the tumor microenvironment. To allow stratification of patients according to tumor GD2 expression, we established and validated immunofluorescence detection of GD2 on paraffin-embedded tumor tissues. Results: Lentiviral all-in-one vector engineering of human T cells with the GD2-specific CAR with and without inducible IL18 resulted in cell products with comparable proportions of CAR-expressing central memory T cells. Production of IL18 strictly depends on GD2 antigen engagement. GD2IL18CART respond to interaction with GD2-positive tumor cells with higher IFNγ and TNFα cytokine release and more effective target cytolysis compared with CAR T cells without inducible IL18. GD2IL18CART further have superior in vivo antitumor activity, with eradication of GD2-positive tumor xenografts. Finally, we established GMP-compliant manufacturing of GD2IL18CART and found it to be feasible and efficient at clinical scale. Conclusions: These results pave the way for clinical investigation of GD2IL18CART in pediatric and adult patients with neuroblastoma and other GD2-positive cancers (EU CT 2022–501725–21–00).

中文翻译：

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CAR T 细胞的临床前开发与抗原诱导 IL18 执行治疗 GD2 阳性实体癌

目的：嵌合抗原受体重定向 T 细胞（CAR T 细胞）的细胞因子工程是克服经典 CAR T 细胞对抗实体癌的有限活性的一个有前景的原理。实验设计：我们开发了一种研究性药物产品 GD2IL18CART，由针对神经节苷脂 GD2 的 CAR T 细胞和 CAR 诱导的 IL18 组成，以增强其在肿瘤微环境中的激活反应和细胞溶解效应功能。为了根据肿瘤 GD2 表达对患者进行分层，我们建立并验证了石蜡包埋肿瘤组织上 GD2 的免疫荧光检测。结果：使用 GD2 特异性 CAR（含或不含诱导型 IL18）对人 T 细胞进行慢病毒一体化载体工程，产生具有可比比例的表达 CAR 的中央记忆 T 细胞的细胞产品。 IL18 的产生严格依赖于 GD2 抗原的结合。与没有诱导型 IL18 的 CAR T 细胞相比，GD2IL18CART 对与 GD2 阳性肿瘤细胞的相互作用做出反应，具有更高的 IFNγ 和 TNFα 细胞因子释放和更有效的靶细胞溶解作用。 GD2IL18CART 还具有优异的体内抗肿瘤活性，可根除 GD2 阳性肿瘤异种移植物。最后，我们建立了符合 GMP 的 GD2IL18CART 生产，并发现其在临床规模上是可行且高效的。结论：这些结果为 GD2IL18CART 在神经母细胞瘤和其他 GD2 阳性癌症儿童和成人患者中的临床研究铺平了道路 (EU CT 2022-501725-21-00)。