抗血管生成肽AP25表面展示细菌外膜囊泡的制备及其肿瘤抑制效应评价

Preparation of bacterial outer membrane vesicles modified with anti-angiogenic peptide AP25 on the surface and evaluation of their anti-tumor effects

摘要:

细菌外膜囊泡(bacterial outer membrane vesicles, OMVs)因其良好的生物相容性、肿瘤穿透性和装载能力，受到纳米载药领域研究人员的广泛关注。AP25肽可以阻断恶性肿瘤血管生成，具有广谱的抗癌活性。为了实现AP25肽的高效递送，本研究通过基因工程技术构建了AP25肽表面展示的细菌外膜囊泡，并探究了其对乳腺癌及胃癌的体外抑制作用。结果表明，工程化OMV具有典型的OMV形貌特征，粒径分布符合理论。蛋白酶K (proteinase K, PK)消化结合Western blotting检测验证AP25肽被修饰在OMV外膜表面。细胞实验显示，WAP25 OMV可以显著抑制MDA-MB-231和HGC-27细胞的增殖、迁移和侵袭，促进细胞凋亡，并且下调肿瘤迁移和血管生成相关蛋白整合素β1 (integrin beta 1, integrin β1)、DNA结合抑制因子1 (Homo sapiens inhibitor of DNA binding 1, ID1)、核因子κB (nuclear factor kappa-B, NF-κB)及血管内皮生长因子(vascular endothelial growth factor, VEGF)的表达。本研究首次实现了蛋白类药物基于OMV的有效递送，为肿瘤抗血管生成治疗和细菌外膜囊泡的功能开发提供了新的思路。

Abstract:

Bacterial outer membrane vesicles (OMVs) have attracted widespread attention in the field of drug delivery due to their excellent biocompatibility, tumor penetration, and loading capacity. The anti-angiogenic peptide AP25 can block malignant tumor angiogenesis and has broad-spectrum anti-cancer activity. To achieve efficient delivery of AP25, we modified AP25 on the surface of OMVs through genetic engineering and explored their inhibitory effects on breast cancer and gastric cancer in vitro. The results indicated that the engineered OMVs had typical morphological characteristics of OMVs, and the particle size distribution conformed to the theoretical. Proteinase K digestion combined with Western blotting confirmed that AP25 was modified on the membrane surface of OMVs. Cell experiments showed that WAP25 OMVs significantly inhibited the proliferation, migration, and invasion of MDA-MB-231 and HGC-27 cells, promoted the cell apoptosis, and downregulated the expression of tumor migration and angiogenesis-related proteins: integrin beta 1 (integrin β1), Homo sapiens inhibitor of DNA binding 1 (ID1), nuclear factor kappa-B (NF-κB), and vascular endothelial growth factor (VEGF). This study achieves effective delivery of protein drugs based on OMVs for the first time, providing new ideas for the anti-angiogenesis therapy for tumors and the functional development of bacterial OMVs.