肿瘤细胞治疗研究
英文原题：Synthesis of mRNA lipid nanoparticles for engineering GD2 CAR T and CAR NK cells against neuroblastoma.
Synthesis of mRNA lipid nanoparticles for engineering GD2 CAR T and CAR NK cells against neuroblastoma.
本研究表明，mRNA LNPs 是一种有前景的非病毒方法，用于生成 GD2 CAR T 和 CAR NK 细胞，可能提供一种比当前基于病毒载体的方法更安全、更具成本效益的替代方案。
用嵌合抗原受体（CAR）工程化T细胞和自然杀伤（NK）细胞，可为癌症治疗创建有效的过继性细胞转移疗法。然而，使用病毒转导作为主要的基因修饰方法会增加监管负担，并且大规模生产成本高昂。通过脂质纳米颗粒（LNP）递送编码CAR的mRNA已被探索作为一种有效的非病毒方法来生成CAR免疫细胞。尽管如此，迄今为止尚未针对神经母细胞瘤的CAR治疗进行优化。设计了一种用于将编码GD2 CAR的mRNA递送至人T细胞和NK细胞的LNP制剂，并通过动态光散射对其粒径分布、z-平均直径、多分散指数和电位进行了表征。mRNA LNP转染后，荧光报告基因检测持续超过1周，且不影响T细胞或NK细胞活力。报告基因阳性率为79.9%的GD2 CAR T细胞和报告基因阳性率为26.6%的GD2 CAR NK细胞的效力在体外针对GD2+神经母细胞瘤细胞系CHLA20进行了评估。GD2 CAR T细胞或CAR NK细胞能够以剂量依赖性方式有效靶向并杀伤神经母细胞瘤细胞，且GD2 CAR T细胞显示出IFN产生增加。本研究表明，mRNA LNP是一种有前景的非病毒方法，用于生成GD2 CAR T细胞和CAR NK细胞，可能提供一种比当前基于病毒载体的方法更安全、更具成本效益的替代方案。
Engineering T and natural killer (NK) cells with chimeric antigen receptors (CAR) creates effective adoptive cell transfer therapies for cancer treatment. However, using viral transduction as a primary genetic modification method adds regulatory burdens and is expensive to produce at scale. Delivering mRNA encoding CAR via lipid nanoparticles (LNPs) has been explored as a potent non-viral method to generate CAR immune cells. Still, it has not been optimized for CAR treatment of neuroblastoma to date. An LNP formulation to deliver mRNA encoding a GD2 CAR into human T and NK cells was designed and characterized by dynamic light scattering for size distribution, z-average diameter, polydispersity index, and potential. Fluorescent reporter detection persisted for more than 1 week after mRNA LNP transfection, without affecting T or NK cell viability. The potency of GD2 CAR T cells with 79.9% reporter positivity and GD2 CAR NK cells with 26.6% reporter positivity was assessed in vitro against the GD2 + neuroblastoma cell line CHLA20. GD2 CAR T or CAR NK cells could effectively target and kill neuroblastoma cells in a dose-dependent fashion, and GD2 CAR T cells showed increased IFN production. This study shows mRNA LNPs are a promising non-viral approach for generating GD2 CAR T and CAR NK cells, potentially offering a safer and more cost-effective alternative to current viral vector-based methods.
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