Unit of Molecular and Genetic Therapeutics, Laboratory of Medical Innovation, Institute for Advanced Medical Sciences, Hyogo College of Medicine, Nishinomiya, Japan.
Division of Pathology and Cell Therapy, Chiba Cancer Center Research Institute, Chiba, Japan.
Cancer Gene Ther. 2019 May;26(5-6):128-135. doi: 10.1038/s41417-018-0051-0. Epub 2018 Oct 22.
Retroviral replicating vectors (RRVs) have been shown to achieve efficient tumor transduction and enhanced therapeutic benefits in a variety of cancer models. In the present study, we evaluated a possible combinatorial effect of prodrug activator genes delivered by two different RRVs derived from amphotropic murine leukemia virus (AMLV) and gibbon ape leukemia virus (GALV) on human hepatocellular carcinoma Hep3B cells. Both RRVs showed efficient replicative spread in culture and can overcame superinfection resistance each other. Notably, the replication and spread of each RRV in culture remained unaffected by pretransduction with the counterpart RRV. We further transduced cells with RRVs which individually possessed the prodrug activator genes yeast cytosine deaminase (CD) and herpes simplex virus thymidine kinase (TK) alone or in combination, and evaluated the cytotoxic effects of RRV-mediated gene therapy with CD and TK in the presence of the respective prodrugs, 5-fluorocytosine and ganciclovir. All combinations of the two prodrug activator genes produced synergistic cytocidal effects, but the combined effects of the different genes were significantly greater than those of the same genes when delivered by two different vectors. The present findings indicate the potential utility of dual-vector gene therapy using two different RRVs carrying different prodrug activator genes.
逆转录病毒复制载体（RRV）已被证明可在多种癌症模型中实现高效的肿瘤转导和增强的治疗效果。在本研究中，我们评估了两种不同的源自嗜性鼠白血病病毒（AMLV）和长臂猿白血病病毒（GALV）的 RRV 传递的前药激活基因的组合效应，对人肝癌 Hep3B 细胞可能产生的影响。两种 RRV 在培养中均显示出有效的复制性扩散，并且可以相互克服超感染抗性。值得注意的是，每种 RRV 在培养中的复制和扩散均不受与对照 RRV 预先转导的影响。我们进一步用单独携带前药激活基因酵母胞嘧啶脱氨酶（CD）和单纯疱疹病毒胸苷激酶（TK）的 RRV 转导细胞，或者将这两种基因组合在一起，并用各自的前体药物 5-氟胞嘧啶和更昔洛韦评估 RRV 介导的基因治疗的细胞毒性作用。两种前药激活基因的所有组合均产生协同细胞杀伤作用，但当由两种不同的载体传递时，不同基因的组合效果明显大于相同基因的组合效果。本研究结果表明，使用携带不同前药激活基因的两种不同 RRV 进行双重载体基因治疗具有潜在的应用价值。
