Zhang Chunhe, Hou Jihuan, Li Zhen, Shen Quan, Bai Haiqing, Chen Li, Shen Jinying, Wang Ping, Su Yinlei, Li Jing, Zhang Qisi, Liu Chengyao, Xi Xuetong, Qi Fei, Chen Yuting, Xie Xin, Ye Adam Yongxin, Liu Xiaoheng, Plebani Roberto, Church George, Si Longlong
State Key Laboratory of Quantitative Synthetic Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
University of Chinese Academy of Sciences, Beijing, China.
Nat Chem Biol. 2025 Jan 15. doi: 10.1038/s41589-024-01813-z.
Manipulating viral protein stability using the cellular ubiquitin-proteasome system (UPS) represents a promising approach for developing live-attenuated vaccines. The first-generation proteolysis-targeting (PROTAR) vaccine had limitations, as it incorporates proteasome-targeting degrons (PTDs) at only the terminal ends of viral proteins, potentially restricting its broad application. Here we developed the next-generation PROTAR vaccine approach, referred to as PROTAR 2.0, which enabled flexible incorporation of PTDs at various genomic loci of influenza viruses, including internal regions and terminal ends. The PROTAR 2.0 influenza viruses maintained efficient replication in UPS-deficient cells for large-scale production but were attenuated by PTD-mediated proteasomal degradation of viral proteins in conventional cells. Incorporation of multiple PTDs into one virus generated optimized PROTAR 2.0 vaccine candidates. In animal models, PROTAR 2.0 vaccine candidates were highly attenuated and a single-dose intranasal immunization induced robust and broad immune responses that provided complete cross-reactive protection against both homologous and heterologous viral challenges.
利用细胞泛素-蛋白酶体系统（UPS）来操控病毒蛋白稳定性，是开发减毒活疫苗的一种很有前景的方法。第一代蛋白酶解靶向（PROTAR）疫苗存在局限性，因为它仅在病毒蛋白的末端掺入蛋白酶体靶向降解子（PTD），这可能会限制其广泛应用。在此，我们开发了下一代PROTAR疫苗方法，即PROTAR 2.0，它能够在流感病毒的各个基因组位点灵活掺入PTD，包括内部区域和末端。PROTAR 2.0流感病毒在缺乏UPS的细胞中能高效复制以进行大规模生产，但在传统细胞中会因PTD介导的病毒蛋白蛋白酶体降解而减毒。将多个PTD掺入一种病毒可产生优化的PROTAR 2.0候选疫苗。在动物模型中，PROTAR 2.0候选疫苗高度减毒，单剂量鼻内免疫可诱导强烈且广泛的免疫反应，能提供针对同源和异源病毒攻击的完全交叉反应性保护。
