Shashkova Elena V, Cherenova Lubov V, Kazansky Dmitry B, Doronin Konstantin
Institute of Agricultural Biotechnology, Moscow, Russia.
Cancer Gene Ther. 2005 Jul;12(7):617-26. doi: 10.1038/sj.cgt.7700822.
Avian adenovirus CELO is a novel adenovirus vector system with the advantages of efficient production, high virion stability, and the absence of crossreactivity with Ad5-neutralizing antibodies. In this study, we evaluated the anticancer efficacy of a CELO vector encoding the herpes simplex virus type 1 thymidine kinase, a prodrug-activating therapeutic gene. Vectors carrying the gene for HSV-tk or EGFP under the control of the HCMV promoter in place of the "nonessential" region of the CELO genome were constructed. Anticancer activity of the CELO-TK vector was studied in vitro, in human and murine tumor cells in cell culture, and in vivo, in established subcutaneous murine B16 melanoma tumors in C57BL/6 mice. The CELO-TK vector mediated delivery of functional HSV-tk to tumor cell lines in cell culture. Comparison of the CELO-TK vector to a first-generation human adenovirus type 5 vector Ad5-TK in cultured H1299 cells showed equal levels of functional activity at increasing multiplicities of infection with CELO-based vector. CELO vectors allowed for transduction and expression of EGFP and HSV-tk genes in subcutaneous melanoma tumors in C57BL/6 mice. Intratumoral injections of CELO-TK followed by ganciclovir administration resulted in suppression of tumor growth and significantly increased the median of survival. The results of the study demonstrated the efficacy of CELO vector as a vehicle for the delivery of prodrug-activating genes such as HSV-tk to tumor cells in vitro and in vivo.
禽腺病毒CELO是一种新型腺病毒载体系统,具有生产效率高、病毒粒子稳定性高以及与Ad5中和抗体无交叉反应等优点。在本研究中,我们评估了一种编码单纯疱疹病毒1型胸苷激酶(一种前药激活治疗基因)的CELO载体的抗癌效果。构建了在CELO基因组的“非必需”区域替换为受HCMV启动子控制的携带HSV-tk或EGFP基因的载体。在体外细胞培养中的人源和鼠源肿瘤细胞以及体内C57BL/6小鼠中已建立的皮下B16黑色素瘤肿瘤中研究了CELO-TK载体的抗癌活性。CELO-TK载体在细胞培养中介导功能性HSV-tk传递至肿瘤细胞系。在培养的H1299细胞中将CELO-TK载体与第一代人5型腺病毒载体Ad5-TK进行比较,结果显示在基于CELO的载体感染复数增加时,两者的功能活性水平相当。CELO载体能够在C57BL/6小鼠的皮下黑色素瘤肿瘤中转导并表达EGFP和HSV-tk基因。瘤内注射CELO-TK后给予更昔洛韦可抑制肿瘤生长并显著提高中位生存期。该研究结果证明了CELO载体作为将前药激活基因(如HSV-tk)递送至体外和体内肿瘤细胞的载体的有效性。