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High-Affinity Decoy PD-1 Mutant Screened from an Epitope-Specific Cell Library
Engineering ( IF 10.1 ) Pub Date : 2021-07-13 , DOI: 10.1016/j.eng.2020.11.011
Hao Liu 1, 2 , Chunxia Qiao 1 , Naijing Hu 1, 3 , Zhihong Wang 1, 4 , Jing Wang 1 , Jiannan Feng 1 , Beifen Shen 1 , Yuanfang Ma 2 , Longlong Luo 1
Affiliation  

Immunotherapy with anti-programmed cell death protein-1 (PD-1)/programmed cell death ligand-1 (PD-L1) monoclonal antibodies has become routine in the treatment of many kinds of human cancers, such as lung cancer, intestinal cancer, and melanoma. The PD-1/PD-L1 pathway inhibits T cell activation in the micro-environment, making it an attractive target against cancer. Wild-type (WT) PD-1 ectodomain has been shown to have difficulty blocking PD-1/PD-L1 mixture formation due to its low affinity. The present work uses three-dimensional (3D) crystal complex structures to analyze the interaction by which PD-1 binds to PD-L1 or PD-L2. It also reports on a theoretical study of the binding mode between PD-1 and its clinical antibody Opdivo. Based on the theoretical binding analysis of PD-1 and its ligands (i.e., PD-L1 and PD-L2) or antibody (Opdivo), a small-content, epitope-oriented mammalian cell library was established for PD-1. After three rounds of cell sorting, the decoy PD-1 mutant 463, which presented a higher affinity than WT PD-1 to the PD-L1 (the affinity has increased by almost three orders of magnitude) was screened out. It exhibited an inhibitory effect against PD-1 to prevent it from forming mixtures with PD-L1, which was similar to the effect of the commercial anti-PD-L1 antibody atezolizumab (ATE). The median effective concentration (EC50) value of the decoy mutant was 0.031 μg·mL−1 in comparison with 0.063 μg·mL−1 for ATE; both values were much lower than that of WT PD-1, at 2.571 μg·mL−1. The 463 decoy mutant reversed the inhibitory function of PD-1 in T cell activation; furthermore, 10 mg·kg−1 of 463 inhibited about 75% of tumor growth in vivo in a MC38 transgenic xenograft mice model, which was similar to the activity of ATE. More interestingly, an even lower dose of 463 (2 mg·kg−1) showed a better effect than 10 mg·kg−1 of WT PD-1. This work offers the decoy 463 with an improved curative effect, which holds potential to become a good option against PD-1/PD-L1-related cancers.



中文翻译:

从表位特异性细胞库中筛选出的高亲和力诱饵 PD-1 突变体

抗程序性细胞死亡蛋白-1(PD-1)/程序性细胞死亡配体-1(PD-L1)单克隆抗体免疫治疗已成为多种人类癌症治疗的常规治疗,如肺癌、肠癌、和黑色素瘤。PD-1/PD-L1 通路抑制微环境中的 T 细胞活化,使其成为抗癌的有吸引力的靶点。已显示野生型 (WT) PD-1 胞外域由于其低亲和力而难以阻断 PD-1/PD-L1 混合物的形成。目前的工作使用三维 (3D) 晶体复合结构来分析 PD-1 与 PD-L1 或 PD-L2 结合的相互作用。它还报告了 PD-1 与其临床抗体 Opdivo 之间结合模式的理论研究。基于PD-1及其配体(即PD-L1和PD-L2)或抗体(Opdivo)的理论结合分析,为 PD-1 建立了一个小内容、以表位为导向的哺乳动物细胞文库。经过三轮细胞分选,筛选出诱饵PD-1突变体463,其对PD-L1表现出比WT PD-1更高的亲和力(亲和力提高了近三个数量级)。它表现出对 PD-1 的抑制作用,防止其与 PD-L1 形成混合物,这类似于商业抗 PD-L1 抗体 atezolizumab (ATE) 的作用。中位有效浓度(EC 它表现出对 PD-1 的抑制作用,防止其与 PD-L1 形成混合物,这类似于商业抗 PD-L1 抗体 atezolizumab (ATE) 的作用。中位有效浓度(EC 它表现出对 PD-1 的抑制作用,防止其与 PD-L1 形成混合物,这类似于商业抗 PD-L1 抗体 atezolizumab (ATE) 的作用。中位有效浓度(EC50 ) 诱饵突变体的值为 0.031 μg·mL -1,而 ATE 的值为0.063 μg·mL -1;这两个值都远低于 WT PD-1 的值,为 2.571 μg·mL -1。463诱饵突变体逆转了PD-1在T细胞活化中的抑制作用;此外,在 MC38 转基因异种移植小鼠模型中,463 中的10 mg·kg -1抑制了约 75% 的体内肿瘤生长,这与 ATE 的活性相似。更有趣的是,甚至更低剂量的 463 (2 mg·kg -1 ) 显示出比 10 mg·kg -1更好的效果WT PD-1。这项工作为诱饵 463 提供了更好的疗效,有可能成为对抗 PD-1/PD-L1 相关癌症的良好选择。

更新日期:2021-07-13
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