Department of Cellular & Molecular Pharmacology, San Francisco, CA, USA.
Center for Systems and Synthetic Biology, University of California San Francisco, San Francisco, 94158, CA, USA.
Sci Rep. 2017 Apr 7;7(1):737. doi: 10.1038/s41598-017-00462-8.
Immunotherapies with chimeric antigen receptor (CAR) T cells and checkpoint inhibitors (including antibodies that antagonize programmed cell death protein 1 [PD-1]) have both opened new avenues for cancer treatment, but the clinical potential of combined disruption of inhibitory checkpoints and CAR T cell therapy remains incompletely explored. Here we show that programmed death ligand 1 (PD-L1) expression on tumor cells can render human CAR T cells (anti-CD19 4-1BBζ) hypo-functional, resulting in impaired tumor clearance in a sub-cutaneous xenograft model. To overcome this suppressed anti-tumor response, we developed a protocol for combined Cas9 ribonucleoprotein (Cas9 RNP)-mediated gene editing and lentiviral transduction to generate PD-1 deficient anti-CD19 CAR T cells. Pdcd1 (PD-1) disruption augmented CAR T cell mediated killing of tumor cells in vitro and enhanced clearance of PD-L1+ tumor xenografts in vivo. This study demonstrates improved therapeutic efficacy of Cas9-edited CAR T cells and highlights the potential of precision genome engineering to enhance next-generation cell therapies.
嵌合抗原受体 (CAR) T 细胞和检查点抑制剂(包括拮抗程序性细胞死亡蛋白 1 [PD-1] 的抗体)的免疫疗法都为癌症治疗开辟了新途径,但联合破坏抑制性检查点和 CAR T 细胞疗法的临床潜力仍未得到充分探索。在这里,我们表明肿瘤细胞上的程序性死亡配体 1 (PD-L1) 的表达可以使人类 CAR T 细胞(抗-CD19 4-1BBζ)功能低下,导致皮下异种移植模型中的肿瘤清除受损。为了克服这种受抑制的抗肿瘤反应,我们开发了一种联合 Cas9 核糖核蛋白 (Cas9 RNP) 介导的基因编辑和慢病毒转导的方案,以生成 PD-1 缺失的抗-CD19 CAR T 细胞。Pdcd1(PD-1)的破坏增强了 CAR T 细胞对肿瘤细胞的体外杀伤作用,并增强了体内 PD-L1+肿瘤异种移植物的清除。这项研究证明了 Cas9 编辑的 CAR T 细胞的治疗效果得到了改善,并强调了精确基因组工程增强下一代细胞疗法的潜力。
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