抗 PSMA CAR NK-92 细胞与抗 PD-L1 单克隆抗体联合治疗增强了对去势抵抗性前列腺癌的抗肿瘤疗效。
Combined treatment with anti-PSMA CAR NK-92 cell and anti-PD-L1 monoclonal antibody enhances the antitumour efficacy against castration-resistant prostate cancer.
机构信息
State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Department of Urology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
出版信息
Clin Transl Med. 2022 Jun;12(6):e901. doi: 10.1002/ctm2.901.
BACKGROUND
The chimeric antigen receptor NK-92 (CAR NK-92) cell targeting the prostate-specific membrane antigen (PSMA) has shown antitumour effects in castration-resistant prostate cancer (CRPC). However, the expression changes of programmed death ligand 1 (PD-L1) and its mechanisms on CAR NK-92 and CRPC cells and the effect of the anti-PD-L1 monoclonal antibody (mAb) on PD-L1 expressed on CAR NK-92 cells remain unknown.
METHODS
Human dendritic cells and CD8 T cells were acquired from blood samples of healthy donors and cocultured with C4-2 cells. Changes in PD-L1 expression were detected by flow cytometry. Differential gene expressions were investigated by RNA sequence analysis, while the regulation of PD-L1 molecular signaling was explored using western blotting. In vitro cytotoxicity was evaluated using the Cell Counting Kit-8 assay and the bioluminescent intensity (BLI) of green fluorescent protein-labelled C4-2 cells. CRPC growth in vivo was monitored using callipers and BLI in male NOD/SCID mice subcutaneously injected with C4-2 cells and treated intravenously with anti-PD-L1/PD-1 mAb, CAR NK-92 or cocultured CD8 T cells.
RESULTS
Significantly upregulated expression of PD-L1k was observed in cocultured C4-2 and CAR NK-92 cells. In addition, upregulation of PD-L1 expression was dependent on interferon-γ in C4-2 cells, while it was dependent on direct cell-to-cell interaction via the NK group 2 member D/ phosphatidylinositol 3-kinase/AKT pathway in CAR NK-92 cells. The anti-PD-L1 mAb directly acted on PD-L1 expressed on CAR NK-92 cells and augmented the cytotoxicity of CAR NK-92 cells against C4-2 and CRPC cells from one patient in vitro. Anti-PD-L1 mAb significantly enhanced the antitumour effect of CAR NK-92 cells against CRPC cells in vivo when compared to treatment with CAR NK-92 cells or combined with anti-PD-1 mAb in the absence or presence of cocultured CD8 T cells.
CONCLUSION
Combined treatment with CAR NK-92 and anti-PD-L1 mAb improved the antitumour efficacy against CRPC, which is of extraordinary translational value in the clinical treatment of CRPC.
背景
靶向前列腺特异性膜抗原（PSMA）的嵌合抗原受体 NK-92（CAR NK-92）细胞在去势抵抗性前列腺癌（CRPC）中显示出抗肿瘤作用。然而，CAR NK-92 和 CRPC 细胞上程序性死亡配体 1（PD-L1）的表达变化及其机制，以及抗 PD-L1 单克隆抗体（mAb）对 CAR NK-92 细胞上表达的 PD-L1 的影响尚不清楚。
方法
从健康供体的血液样本中获得人树突状细胞和 CD8 T 细胞，并与 C4-2 细胞共培养。通过流式细胞术检测 PD-L1 表达的变化。通过 RNA 序列分析研究差异基因表达，通过 Western blot 研究 PD-L1 分子信号的调节。使用细胞计数试剂盒-8 测定法和绿色荧光蛋白标记的 C4-2 细胞的生物发光强度（BLI）评估体外细胞毒性。通过卡尺和皮下注射 C4-2 细胞的雄性 NOD/SCID 小鼠的 BLI 监测体内 CRPC 的生长，并通过静脉内给予抗 PD-L1/PD-1 mAb、CAR NK-92 或共培养的 CD8 T 细胞进行治疗。
结果
在共培养的 C4-2 和 CAR NK-92 细胞中观察到 PD-L1k 的表达显著上调。此外，PD-L1 表达的上调依赖于 C4-2 细胞中的干扰素-γ，而在 CAR NK-92 细胞中则依赖于通过 NK 组 2 成员 D/磷酸肌醇 3-激酶/AKT 通路的直接细胞间相互作用。抗 PD-L1 mAb 直接作用于 CAR NK-92 细胞上表达的 PD-L1，并增强了 CAR NK-92 细胞对体外 C4-2 和 CRPC 细胞的细胞毒性。与单独使用 CAR NK-92 细胞或在共培养的 CD8 T 细胞存在或不存在的情况下联合使用抗 PD-1 mAb 相比，抗 PD-L1 mAb 显著增强了 CAR NK-92 细胞对体内 CRPC 细胞的抗肿瘤作用。
结论
CAR NK-92 和抗 PD-L1 mAb 的联合治疗改善了对 CRPC 的抗肿瘤疗效，这在 CRPC 的临床治疗中具有特殊的转化价值。
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