Thailand Excellence Centre for Tissue Engineering and Stem Cells, Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
Cancer Phage Therapy Group, Department of Brain Sciences, Faculty of Medicine, Imperial College London, London, UK.
FASEB J. 2021 May;35(5):e21487. doi: 10.1096/fj.202002539R.
Chondrosarcoma is a cartilage-forming bone tumor, well known for intrinsic resistance to chemotherapy and radiotherapy. We have designed a targeted chondrosarcoma gene therapy using a bacteriophage (phage) particle to deliver therapeutic genes. Phage has no tropism for mammalian cells, allowing engineered phage to be targeted to specific cell surface receptors in cancer. We modified the phage capsid to display the RGD4C ligand on the pIII minor coat proteins to specifically bind to αvβ3 or αvβ5 integrin receptors. The endosomal escape peptide, H5WYG, was also displayed on recombinant pVIII major coat proteins to enhance gene delivery. Finally, a human tumor necrosis factor alpha (TNFα) therapeutic transgene expression cassette was incorporated into the phage genome. First, we found that human chondrosarcoma cells (SW1353) have high expression of αvβ3, αvβ5 integrin receptors, and both TNFα receptors. Targeted particle encoding a luciferase reporter gene efficiently and selectively mediated gene delivery to these cells. When SW1353 cells were treated with the targeted particle encoding a TNFα transgene, significant cell killing was evident and was associated with high expression of TNFα and apoptosis-related genes. In vivo, mice with established human chondrosarcoma showed suppression of tumors upon repetitive intravenous administrations of the targeted phage. These data show that our phage-based particle is a promising, selective, and efficient tool for targeted chondrosarcoma therapy.
软骨肉瘤是一种软骨形成的骨肿瘤，以对化疗和放疗的固有耐药性而闻名。我们设计了一种使用噬菌体（phage）颗粒进行靶向软骨肉瘤基因治疗的方法，以传递治疗基因。噬菌体对哺乳动物细胞没有亲嗜性，允许工程噬菌体靶向癌症中特定的细胞表面受体。我们修饰了噬菌体衣壳，在 pIII 次要衣壳蛋白上展示 RGD4C 配体，以特异性结合αvβ3 或αvβ5 整合素受体。还在重组 pVIII 主要衣壳蛋白上展示了内体逃逸肽 H5WYG，以增强基因传递。最后，将人肿瘤坏死因子α（TNFα）治疗性转基因表达盒整合到噬菌体基因组中。首先，我们发现人软骨肉瘤细胞（SW1353）高表达αvβ3、αvβ5 整合素受体以及两种 TNFα 受体。编码荧光素酶报告基因的靶向颗粒有效地、选择性地介导了这些细胞的基因传递。当用编码 TNFα 转基因的靶向颗粒处理 SW1353 细胞时，明显观察到细胞杀伤，并且与 TNFα 和与凋亡相关的基因的高表达相关。在体内，荷有人软骨肉瘤的小鼠在重复静脉注射靶向噬菌体后肿瘤受到抑制。这些数据表明，我们基于噬菌体的颗粒是一种有前途的、选择性的、有效的靶向软骨肉瘤治疗工具。
