中文摘要：软骨肉瘤是一种形成软骨骨肿瘤，对化疗和放疗具有固有耐药性。本研究设计了一种靶向软骨肉瘤基因疗法，使用噬菌体颗粒传递治疗基因。噬菌体对哺乳动物细胞没有趋性，所以工程噬菌体能够靶向肿瘤细胞表面的特异性受体。首先修饰噬菌体衣壳，在pIII小外壳蛋白上展示RGD4C配体，以特异性结合αvβ3或αvβ5整合素受体。再将内体逃逸肽H5WYG也展示在重组pVIII主要外壳蛋白上，以增强基因传递。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。研究发现，人类软骨肉瘤细胞(SW1353)可高表达αvβ3、αvβ5整合素受体和两种TNF-α受体。编码荧光素酶报告基因的靶向粒子有效且选择性地介导基因递送到这些细胞。当用编码TNF-α转基因的靶向颗粒处理SW1353细胞时，细胞杀伤作用很明显，并且与TNF-α和凋亡相关基因的高表达相关。在体内，经反复静脉注射靶向噬菌体后，已建立的人软骨肉瘤小鼠显示出肿瘤抑制作用。可见，噬菌体颗粒是一种有前途的、选择性的、有效的软骨肉瘤靶向治疗工具。
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
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