Department of Neurology, Clinical Neuroscience Center, University Hospital Zurich and University of Zurich, Zurich, Switzerland.
Department of Dermatology, University Hospital Zurich and University of Zurich, Zurich, Switzerland.
Clin Cancer Res. 2022 Nov 1;28(21):4747-4756. doi: 10.1158/1078-0432.CCR-21-4384.
Most chimeric antigen receptor (CAR) T-cell strategies against glioblastoma have demonstrated only modest therapeutic activity and are based on persistent gene modification strategies that have limited transgene capacity, long manufacturing processes, and the risk for uncontrollable off-tumor toxicities. mRNA-based T-cell modifications are an emerging safe, rapid, and cost-effective alternative to overcome these challenges, but are underexplored against glioblastoma.
We generated mouse and human mRNA-based multifunctional T cells coexpressing a multitargeting CAR based on the natural killer group 2D (NKG2D) receptor and the proinflammatory cytokines IL12 and IFNα2 and assessed their antiglioma activity in vitro and in vivo.
Compared with T cells that either expressed the CAR or cytokines alone, multifunctional CAR T cells demonstrated increased antiglioma activity in vitro and in vivo in three orthotopic immunocompetent mouse glioma models without signs of toxicity. Mechanistically, the coexpression of IL12 and IFNα2 in addition to the CAR promoted a proinflammatory tumor microenvironment and reduced T-cell exhaustion as demonstrated by ex vivo immune phenotyping, cytokine profiling, and RNA sequencing. The translational potential was demonstrated by image-based single-cell analyses of mRNA-modified T cells in patient glioblastoma samples with a complex cellular microenvironment. This revealed strong antiglioma activity of human mRNA-based multifunctional NKG2D CAR T cells coexpressing IL12 and IFNα2 whereas T cells that expressed either the CAR or cytokines alone did not demonstrate comparable antiglioma activity.
These data provide a robust rationale for future clinical studies with mRNA-based multifunctional CAR T cells to treat malignant brain tumors.
大多数针对胶质母细胞瘤的嵌合抗原受体 (CAR) T 细胞策略仅显示出适度的治疗活性，并且基于持久性基因修饰策略，这些策略具有有限的转基因容量、漫长的制造过程和不可控的肿瘤外毒性风险。基于 mRNA 的 T 细胞修饰是一种新兴的安全、快速且具有成本效益的替代方法，可以克服这些挑战，但针对胶质母细胞瘤的研究还很有限。
我们生成了表达基于自然杀伤组 2D (NKG2D) 受体和促炎细胞因子 IL12 和 IFNα2 的多靶向 CAR 的基于 mRNA 的多功能 T 细胞，评估了它们在体外和体内的抗神经胶质瘤活性。
与单独表达 CAR 或细胞因子的 T 细胞相比，多功能 CAR T 细胞在三种原位免疫活性小鼠神经胶质瘤模型中表现出增强的体外和体内抗神经胶质瘤活性，且没有毒性迹象。从机制上讲，CAR 共表达 IL12 和 IFNα2 除了促进促炎肿瘤微环境外，还减少了 T 细胞衰竭，这一点通过体外免疫表型分析、细胞因子谱分析和 RNA 测序得到证明。通过对具有复杂细胞微环境的患者胶质母细胞瘤样本进行基于图像的单细胞分析，证明了 mRNA 修饰的多功能 NKG2D CAR T 细胞的翻译潜力。这揭示了共表达 IL12 和 IFNα2 的人源基于 mRNA 的多功能 NKG2D CAR T 细胞具有强烈的抗神经胶质瘤活性，而单独表达 CAR 或细胞因子的 T 细胞则没有表现出可比的抗神经胶质瘤活性。
这些数据为未来使用基于 mRNA 的多功能 CAR T 细胞治疗恶性脑肿瘤的临床研究提供了强有力的依据。
