Chimeric antigen receptor (CAR)-T cell therapy has shown limited success in patients with solid tumors. Recent in vitro and in vivo data have shown that adrenoceptor beta-2 (ADRB2) is a novel checkpoint receptor that inhibits T cell-mediated anti-tumor responses. To inhibit ADRB2-mediated inhibitory signaling, we downregulated ADRB2 in CAR-T (shβ2-CAR-T) cells via RNA interference, assessed different parameters, and compared them with conventional second-generation CAR-T cells. ADRB2 knockdown CAR-T cells exhibited enhanced cytotoxicity against prostate cancer cell lines in vitro, by increasing CD69, CD107a, GzmB, IFN-γ, T-bet, and GLUT-1. In addition, ADRB2 deficiency led to improved proliferation, increased CD8/CD4 T cell ratio, and decreased apoptosis in CAR-T cells. shβ2-CAR-T cells expressed more Bcl-2 and led to the generation of more significant proportions of T central memory cells. Finally, the ZAP-70/NF-κB signaling axis was shown to be responsible for the improved functions of novel CAR-T cells. In tumor-bearing mice, shβ2-CAR-T cells performed better than conventional CAR-T cells in eradicating prostate tumors. The study provides the basis for future clinical and translational CAR-T cell research to focus on adrenergic stress-mediated challenges in the tumor microenvironment of stressed tumors.

中文翻译：

---

内源性 ADRB2 抑制提高了 CAR-T 细胞疗法对前列腺癌的疗效


嵌合抗原受体 （CAR）-T 细胞疗法在实体瘤患者中的成功率有限。 最近的体外和 体内数据表明，肾上腺素能受体 β-2 （ADRB2） 是一种新型检查点受体，可抑制 T 细胞介导的抗肿瘤反应。为了抑制 ADRB2 介导的抑制信号传导，我们通过 RNA 干扰下调 CAR-T （shβ2-CAR-T） 细胞中的 ADRB2，评估不同的参数，并将其与传统的第二代 CAR-T 细胞进行比较。ADRB2 敲低 CAR-T 细胞通过 增加 CD69 、 CD107a 、 GzmB 、 IFN-γ 、 T-bet 和 GLUT-1 在体外表现出增强的对前列腺癌细胞系的细胞毒性。此外，ADRB2 缺陷导致 CAR-T 细胞增殖改善，CD8/CD4 T 细胞比率增加，细胞凋亡减少。shβ2-CAR-T 细胞表达更多的 Bcl-2 并导致产生更显着比例的 T 中枢记忆细胞。最后，ZAP-70/NF-κB 信号轴被证明负责改善新型 CAR-T 细胞的功能。在荷瘤小鼠中，shβ2-CAR-T 细胞在根除前列腺肿瘤方面表现优于常规 CAR-T 细胞。该研究为未来的临床和转化 CAR-T 细胞研究提供了基础，以专注于应激肿瘤肿瘤微环境中肾上腺素能应激介导的挑战。