Glypican-3-targeted macrophages delivering drug-loaded exosomes offer efficient cytotherapy in mouse models of solid tumours

Affiliations

• NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, 44 Wenhuaxi Road, 250012, Jinan, Shandong Province, China.
• NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, 44 Wenhuaxi Road, 250012, Jinan, Shandong Province, China. zhangnancy9@sdu.edu.cn.
• NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, 44 Wenhuaxi Road, 250012, Jinan, Shandong Province, China. liuyongjun@sdu.edu.cn.

PMID:  39313508  DOI: 10.1038/s41467-024-52500-5

Abstract

Cytotherapy is a strategy to deliver modified cells to a diseased tissue, but targeting solid tumours remains challenging. Here we design macrophages, harbouring a surface glypican-3-targeting peptide and carrying a cargo to combat solid tumours. The anchored targeting peptide facilitates tumour cell recognition by the engineered macrophages, thus enhancing specific targeting and phagocytosis of tumour cells expressing glypican-3. These macrophages carry a cargo of the TLR7/TLR8 agonist R848 and INCB024360, a selective indoleamine 2,3-dioxygenase 1 (IDO1) inhibitor, wrapped in C16-ceramide-fused outer membrane vesicles (OMV) of Escherichia coli origin (RILO). The OMVs facilitate internalization through caveolin-mediated endocytosis, and to maintain a suitable nanostructure, C16-ceramide induces membrane invagination and exosome generation, leading to the release of cargo-packed RILOs through exosomes. RILO-loaded macrophages exert therapeutic efficacy in mice bearing H22 hepatocellular carcinomas, which express high levels of glypican-3. Overall, we lay down the proof of principle for a cytotherapeutic strategy to target solid tumours and could complement conventional treatment.

细胞疗法是一种向病变组织输送改良细胞的策略，但针对实体瘤的细胞疗法仍具有挑战性。在这里，我们设计了巨噬细胞，其表面含有glypican-3靶向肽，并携带一种抗实体瘤的货物。锚定的靶向肽有助于工程巨噬细胞识别肿瘤细胞，从而增强对表达 glypican-3 的肿瘤细胞的特异性靶向和吞噬作用。这些巨噬细胞携带的货物包括 TLR7/TLR8 激动剂 R848 和 INCB024360（一种选择性吲哚胺 2,3-二氧合酶 1（IDO1）抑制剂），包裹在源于大肠杆菌（RILO）的 C16-神经酰胺融合外膜囊泡（OMV）中。OMV通过窖蛋白介导的内吞作用促进内化，为了保持合适的纳米结构，C16-神经酰胺诱导膜内陷和外泌体生成，从而通过外泌体释放出装有货物的 RILO。装载了 RILO 的巨噬细胞对携带高水平 glypican-3 的 H22 肝细胞癌小鼠具有疗效。总之，我们证明了针对实体瘤的细胞治疗策略的原理，并可作为常规治疗的补充。