肿瘤细胞治疗研究
英文原题：Natural killer cell-specific chimeric antigen receptor enhances CAR NK cell functions and anti-tumor activity.
背景：与 T 细胞不同，自然杀伤（NK）细胞缺乏一个类似于 T 细胞受体（TCR）的、主导其激活的优势激活受体。
背景：与T细胞不同，自然杀伤（NK）细胞缺乏类似控制其激活的T细胞受体（TCR）的主导性激活受体。专门为T细胞设计的嵌合抗原受体（CAR）构建体是否能有效驱动NK细胞激活仍未解决。NK细胞固有地具有非特异性识别能力，并对多种肿瘤靶标发挥广谱细胞毒性。然而，CAR NK细胞与易感靶细胞之间受体-配体相互作用的复杂性阻碍了阐明单个CAR构建体具体功能贡献的努力。 方法：CAR NK细胞通过电穿孔生成。使用慢病毒转导对小鼠B16黑色素瘤细胞系进行改造，以表达多种靶蛋白。采用体外功能测定，包括结合物形成、颗粒极化、脱颗粒、细胞毒性和细胞因子产生，以评估CAR NK细胞的效力。使用重组蛋白包被的微珠研究下游激活信号通路。使用NPG小鼠异种移植模型评估CAR NK细胞的体内抗肿瘤活性。 结果：B16细胞系首先被验证为适合特异性评估CAR NK细胞中CAR构建体功能的模型。在生成的九种不同CAR分子中，包含NKG2DTM-2B4-FCER1G的构建体表现出最强的增强NK细胞介导功能的能力。与这些功能改善一致，该CAR构建体诱导了关键激活通路的强烈磷酸化，包括AKT、VAV1、ERK、PLCγ1和NF-κB。结论：包含 NKG2DTM-2B4-FCER1G 的 CAR 构建体被证明在增强 NK 细胞功能方面最为有效。
Background : Unlike T cells, natural killer (NK) cells lack a dominant activating receptor analogous to the T cell receptor (TCR) that governs their activation. Whether chimeric antigen receptor (CAR) constructs engineered specifically for T cells can effectively drive NK cell activation remains unresolved. NK cells inherently possess non-specific recognition capacities and exert broad-spectrum cytotoxicity against diverse tumor targets. However, the complexity of receptor-ligand interactions between CAR NK cells and susceptible target cells has impeded efforts to delineate the specific functional contributions of individual CAR constructs. Methods : CAR NK cells were generated via electroporation. The murine B16 melanoma cell line was modified to express various target proteins using lentiviral transduction. In vitro functional assays, including conjugate formation, granule polarization, degranulation, cytotoxicity, and cytokine production, were employed to assess CAR NK cell efficacy. Recombinant protein-coated beads were used to investigate downstream activation signaling pathways. The in vivo antitumor activity of CAR NK cells was evaluated using NPG mouse xenograft models. Results : B16 cell line was first validated to be a suitable model for specifically assessing CAR construct function in CAR NK cells. Among nine distinct CAR molecules generated, the construct incorporating the NKG2DTM-2B4-FCER1G exhibited the most potent capacity to enhance NK cell-mediated functionalities. Consistent with these functional improvements, this CAR construct induced robust phosphorylation of key activation pathways, including AKT, VAV1, ERK, PLC 1, and NF- B. Conclusions : The CAR construct incorporating the NKG2DTM-2B4-FCER1G is demonstrated to be the most effective in enhancing NK cell functionality.
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