中文摘要：软骨肉瘤是软骨形成的骨肿瘤，以对化学疗法和放射疗法的内在抗性而被人们熟知。本文设计了靶向软骨肉瘤基因疗法，使用噬菌体颗粒来递送治疗基因。噬菌体对哺乳动物细胞没有嗜性，使得工程噬菌体靶向结合癌症中的特定细胞表面受体成为可能。首先修饰噬菌体衣壳使其在pIII次要外壳蛋白上表达RGD4C配体，以特异性结合αvβ3或αvβ5整合素受体。并使内体逃逸肽H5WYG也在重组pVIII主要外壳蛋白上表达以增强基因递送。最后，将人肿瘤坏死因子-α（TNF-α）治疗性转基因表达盒整合到噬菌体基因组中。首先，人软骨肉瘤细胞（SW1353）高度表达了αvβ3，αvβ5整合素受体和TNFα受体。编码荧光素酶报告基因的靶向颗粒有效且选择性地介导基因递送至这些细胞。当用编码TNF-α转基因的靶向颗粒处理SW1353细胞时，杀伤效果十分明显，并且与TNF-α和凋亡相关基因的高表达相关。在体内，向患有人软骨肉瘤的小鼠静脉内重复施用靶向噬菌体，已显示其对肿瘤的抑制。可见，本研究基于噬菌体的颗粒是有前景、有选择和有效的靶向治疗软骨肉瘤的工具。
外文摘要：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 alpha v beta 3 or alpha v beta 5 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 (TNF alpha) therapeutic transgene expression cassette was incorporated into the phage genome. First, we found that human chondrosarcoma cells (SW1353) have high expression of alpha v beta 3, alpha v beta 5 integrin receptors, and both TNF alpha 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 TNF alpha transgene, significant cell killing was evident and was associated with high expression of TNF alpha 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.
外文关键词：chondrosarcoma；integrins alpha v beta 3 and alpha v beta 5 apoptosis； phage-based particle；targeted gene therapy；tumor necrosis factor alpha(TNF alpha)
作者：Chongchai, A；Waramit, S；Suwan, K；Al-Bahrani, M；Udomruk, S；Phitak, T；Kongtawelert, P；Pothacharoen, P；Hajitou, A
作者单位：Chiang Mai Univ；Imperial Coll London
期刊名称：FASEB JOURNAL
期刊影响因子：4.966
出版年份：2021
出版刊次：5
点击下载：通过选择性递送肿瘤坏死因子-α（TNFα）基因的噬菌体介导的软骨肉瘤疗法