2021年 第7卷 第11期
《工程（英文）》 >> 2021年 第7卷 第11期 doi: 10.1016/j.eng.2020.11.011
基于表位定向细胞库筛选高亲和力PD-1突变体诱骗分子
a State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China
b Joint National Laboratory for Antibody Drug Engineering, the First Affiliated Hospital, School of Medicine, Henan University, Kaifeng 475004, China
c School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang 110016, China
d School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing 210009, China
# These authors contributed equally to this work.
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摘要
抗程序性细胞死亡蛋白-1（programmed cell death protein-1, PD-1）/程序性细胞死亡配体-1（programmed cell death ligand-1, PD-L1）单克隆抗体的免疫疗法已成为治疗肺癌、肠癌和黑色素瘤等多种癌症的常规方法。PD-1/PD-L1信号通路在肿瘤微环境中可抑制T细胞活化，使其成为一个热门的抗癌靶点。野生型（wild type, WT）PD-1胞外结构域由于亲和力低而难以阻断PD-1/PD-L1复合物的形成。本文利用三维（3D）晶体复合结构分析了PD-1与PD-L1或PD-L2的相互作用。文中还报道了PD-1与其临床抗体Opdivo结合模式的理论研究。通过对PD-1及其配体（即PD-L1和PD-L2）或抗体Opdivo的理论结合分析，建立了PD-1的一个小库容量、表位定向的哺乳动物细胞文库。经过三轮细胞分选，筛选出对PD-L1有较高亲和力的PD-1突变体463（亲和力较野生型PD-1提高近3个数量级）。它对PD-1具有抑制作用，可阻止PD-1与PD-L1形成复合物，这与商品化的抗PD-L1抗体atezolizumab（ATE）的作用相似。突变体463的半数有效浓度（median effective concentration, EC50）为0.031 μg·mL−1，而ATE的EC50为0.063 μg·mL−1，两者均明显低于野生型PD-1的EC50 (2.571 μg·mL−1)。在MC38转基因小鼠模型中，463可有效逆转PD-1对T细胞活化的抑制作用，而且10 mg·kg−1的463突变体蛋白对肿瘤生长的抑制率约为75%，与同等剂量下ATE的抑制活性相似。更有趣的是，低剂量的463（2 mg·kg−1）显示出比10 mg·kg−1的野生型PD-1更好的抑瘤效果。这项工作提供了一种高亲和力诱饵骗分子463，其体内外活性较天然PD-1分子有明显提高，因此，它有可能成为靶向PD-1/PD-L1治疗相关肿瘤的良好选择。
关键词
诱饵程序性细胞死亡蛋白-1 ; 程序性细胞死亡配体-1 ; 哺乳动物细胞文库 ; 表位定向
补充材料
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参考文献
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