Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA.
Laboratory for Critical Quality Attributes of Cell Therapy Products, Forevertek Biotechnology, Janshan Road, Changsha Hi-Tech Industrial Development Zone, Changsha 410205, China.
Int J Mol Sci. 2023 Aug 29;24(17):13364. doi: 10.3390/ijms241713364.
Natural killer (NK) cells are cytotoxic lymphocytes that are critical for the innate immune system. Engineering NK cells with chimeric antigen receptors (CARs) allows CAR-NK cells to target tumor antigens more effectively. In this report, we present novel CAR mRNA-LNP (lipid nanoparticle) technology to effectively transfect NK cells expanded from primary PBMCs and to generate functional CAR-NK cells. CD19-CAR mRNA and BCMA-CAR mRNA were embedded into LNPs that resulted in 78% and 95% CAR expression in NK cells, respectively. BCMA-CAR-NK cells after transfection with CAR mRNA-LNPs killed multiple myeloma RPMI8226 and MM1S cells and secreted IFN-gamma and Granzyme B in a dose-dependent manner in vitro. In addition, CD19-CAR-NK cells generated with CAR mRNA-LNPs killed Daudi and Nalm-6 cells and secreted IFN-gamma and Granzyme B in a dose-dependent manner. Both BCMA-CAR-NK and CD19-CAR-NK cells showed significantly higher cytotoxicity, IFN-gamma, and Granzyme B secretion compared with normal NK cells. Moreover, CD19-CAR-NK cells significantly blocked Nalm-6 tumor growth in vivo. Thus, non-viral delivery of CAR mRNA-LNPs can be used to generate functional CAR-NK cells with high anti-tumor activity.
自然杀伤 (NK) 细胞是细胞毒性淋巴细胞，对先天免疫系统至关重要。用嵌合抗原受体 (CAR) 工程化 NK 细胞可使 CAR-NK 细胞更有效地靶向肿瘤抗原。在本报告中，我们提出了一种新型的 CAR mRNA-LNP（脂质纳米颗粒）技术，可有效转染从原代 PBMC 扩增的 NK 细胞，并生成功能性 CAR-NK 细胞。CD19-CAR mRNA 和 BCMA-CAR mRNA 被嵌入 LNP 中，分别导致 NK 细胞中 CAR 表达达到 78%和 95%。用 CAR mRNA-LNP 转染后的 BCMA-CAR-NK 细胞可杀伤多发性骨髓瘤 RPMI8226 和 MM1S 细胞，并在体外以剂量依赖性方式分泌 IFN-γ和 Granzyme B。此外，用 CAR mRNA-LNP 生成的 CD19-CAR-NK 细胞可杀伤 Daudi 和 Nalm-6 细胞，并以剂量依赖性方式分泌 IFN-γ和 Granzyme B。与正常 NK 细胞相比，BCMA-CAR-NK 和 CD19-CAR-NK 细胞均显示出更高的细胞毒性、IFN-γ和 Granzyme B 分泌水平。此外，CD19-CAR-NK 细胞可显著抑制体内 Nalm-6 肿瘤的生长。因此，CAR mRNA-LNP 的非病毒递送可用于生成具有高抗肿瘤活性的功能性 CAR-NK 细胞。
