{"title":"通过基于晚期结构域的工程支架在细胞外囊泡表面展示 αPD-L1 和 αCD3 抗体,以激活 T 细胞抗肿瘤免疫。","authors":"Rui Chen, Ziqin Kang, Wenhao Li, Tianshu Xu, Yongqiang Wang, Qiming Jiang, Yuepeng Wang, Zixian Huang, Xiaoding Xu, Zhiquan Huang","doi":"10.1002/jev2.12490","DOIUrl":null,"url":null,"abstract":"<p>Extracellular vesicles (EVs) are emerging as promising carriers for the delivery of therapeutic biologics. Genetic engineering represents a robust strategy for loading proteins of interest into EVs. Identification of EV-enriched proteins facilitates protein cargo loading efficiency. Many EV-enriched proteins are sorted into EVs via an endosomal sorting complex required for transport (ESCRT)-dependent pathway. In parallel, viruses hijack this EV biosynthesis machinery via conserved late domain motifs to promote egress from host cells. Inspired by the similarity of biogenesis between EVs and viruses, we developed a synthetic, Late domain-based EV scaffold protein that enables the display of a set of single chain variable fragments (scFvs) on the EV surface. We named this scaffold the Late domain-based exosomal antibody surface display platform (LEAP). We applied the LEAP scaffold to reprogramme HEK293T cell-derived EVs to elicit T-cell anti-tumor immunity by simultaneously displaying αPD-L1 and αCD3 scFvs on the EV surface (denoted as αPD-L1×αCD3 bispecific T-cell engaging exosomes, BiTExos). We demonstrated that αPD-L1×αCD3 BiTExos actively redirected T cells to bind to PD-L1<sup>+</sup> tumor cells, promoting T-cell activation, proliferation and tumoricidal cytokine production. Furthermore, the αPD-L1×αCD3 BiTExos promoted T-cell infiltration into the tumor microenvironment to mitigate the tumor burden in vivo. Our study suggested that the LEAP scaffold may serve as a platform for EV surface display and could be applied for a broad range of EV-based biomedical applications.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 7","pages":""},"PeriodicalIF":15.5000,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11270581/pdf/","citationCount":"0","resultStr":"{\"title\":\"Extracellular vesicle surface display of αPD-L1 and αCD3 antibodies via engineered late domain-based scaffold to activate T-cell anti-tumor immunity\",\"authors\":\"Rui Chen, Ziqin Kang, Wenhao Li, Tianshu Xu, Yongqiang Wang, Qiming Jiang, Yuepeng Wang, Zixian Huang, Xiaoding Xu, Zhiquan Huang\",\"doi\":\"10.1002/jev2.12490\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Extracellular vesicles (EVs) are emerging as promising carriers for the delivery of therapeutic biologics. 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We applied the LEAP scaffold to reprogramme HEK293T cell-derived EVs to elicit T-cell anti-tumor immunity by simultaneously displaying αPD-L1 and αCD3 scFvs on the EV surface (denoted as αPD-L1×αCD3 bispecific T-cell engaging exosomes, BiTExos). We demonstrated that αPD-L1×αCD3 BiTExos actively redirected T cells to bind to PD-L1<sup>+</sup> tumor cells, promoting T-cell activation, proliferation and tumoricidal cytokine production. Furthermore, the αPD-L1×αCD3 BiTExos promoted T-cell infiltration into the tumor microenvironment to mitigate the tumor burden in vivo. 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引用次数: 0
摘要
细胞外囊泡(EVs)正在成为输送治疗性生物制剂的理想载体。基因工程是将感兴趣的蛋白质装载到 EVs 中的有力策略。鉴定 EV 富集蛋白有助于提高蛋白质货物的装载效率。许多EV富集蛋白都是通过一种依赖于内体运输所需的分选复合物(ESCRT)的途径分选到EV中的。与此同时,病毒通过保守的晚期结构域基团劫持这一EV生物合成机制,以促进从宿主细胞中排出。受EV和病毒之间相似的生物发生机制的启发,我们开发了一种基于晚期结构域的合成EV支架蛋白,它能在EV表面显示一组单链可变片段(scFvs)。我们将这种支架命名为基于晚期结构域的外泌体抗体表面展示平台(LEAP)。我们应用 LEAP 支架对 HEK293T 细胞衍生的外泌体进行了重编程,通过在外泌体表面同时展示 αPD-L1 和 αCD3 scFvs 来诱导 T 细胞抗肿瘤免疫(称为 αPD-L1×αCD3 双特异性 T 细胞吸引外泌体,BiTExos)。我们证实,αPD-L1×αCD3 BiTExos能主动引导T细胞与PD-L1+肿瘤细胞结合,促进T细胞活化、增殖和杀伤肿瘤细胞因子的产生。此外,αPD-L1×αCD3 BiTExos 还能促进 T 细胞浸润肿瘤微环境,减轻体内肿瘤负荷。我们的研究表明,LEAP支架可作为一种EV表面展示平台,并可广泛应用于基于EV的生物医学领域。
Extracellular vesicle surface display of αPD-L1 and αCD3 antibodies via engineered late domain-based scaffold to activate T-cell anti-tumor immunity
Extracellular vesicles (EVs) are emerging as promising carriers for the delivery of therapeutic biologics. Genetic engineering represents a robust strategy for loading proteins of interest into EVs. Identification of EV-enriched proteins facilitates protein cargo loading efficiency. Many EV-enriched proteins are sorted into EVs via an endosomal sorting complex required for transport (ESCRT)-dependent pathway. In parallel, viruses hijack this EV biosynthesis machinery via conserved late domain motifs to promote egress from host cells. Inspired by the similarity of biogenesis between EVs and viruses, we developed a synthetic, Late domain-based EV scaffold protein that enables the display of a set of single chain variable fragments (scFvs) on the EV surface. We named this scaffold the Late domain-based exosomal antibody surface display platform (LEAP). We applied the LEAP scaffold to reprogramme HEK293T cell-derived EVs to elicit T-cell anti-tumor immunity by simultaneously displaying αPD-L1 and αCD3 scFvs on the EV surface (denoted as αPD-L1×αCD3 bispecific T-cell engaging exosomes, BiTExos). We demonstrated that αPD-L1×αCD3 BiTExos actively redirected T cells to bind to PD-L1+ tumor cells, promoting T-cell activation, proliferation and tumoricidal cytokine production. Furthermore, the αPD-L1×αCD3 BiTExos promoted T-cell infiltration into the tumor microenvironment to mitigate the tumor burden in vivo. Our study suggested that the LEAP scaffold may serve as a platform for EV surface display and could be applied for a broad range of EV-based biomedical applications.
期刊介绍:
The Journal of Extracellular Vesicles is an open access research publication that focuses on extracellular vesicles, including microvesicles, exosomes, ectosomes, and apoptotic bodies. It serves as the official journal of the International Society for Extracellular Vesicles and aims to facilitate the exchange of data, ideas, and information pertaining to the chemistry, biology, and applications of extracellular vesicles. The journal covers various aspects such as the cellular and molecular mechanisms of extracellular vesicles biogenesis, technological advancements in their isolation, quantification, and characterization, the role and function of extracellular vesicles in biology, stem cell-derived extracellular vesicles and their biology, as well as the application of extracellular vesicles for pharmacological, immunological, or genetic therapies.
The Journal of Extracellular Vesicles is widely recognized and indexed by numerous services, including Biological Abstracts, BIOSIS Previews, Chemical Abstracts Service (CAS), Current Contents/Life Sciences, Directory of Open Access Journals (DOAJ), Journal Citation Reports/Science Edition, Google Scholar, ProQuest Natural Science Collection, ProQuest SciTech Collection, SciTech Premium Collection, PubMed Central/PubMed, Science Citation Index Expanded, ScienceOpen, and Scopus.