Systemic cancer therapy with a tumor-selective vaccinia virus mutant lacking thymidine kinase and vaccinia growth factor genes.
Systemic cancer therapy with a tumor-selective vaccinia virus mutant lacking thymidine kinase and vaccinia growth factor genes.
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发表时间：
2001-12
期刊：
影响因子：
11.2
通讯作者：
J. McCart;Jerrold M. Ward;John G. Lee;Yun Hu;H. Alexander;S. Libutti;B. Moss;D. Bartlett
中科院分区：
文献类型：
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作者：
J. McCart;Jerrold M. Ward;John G. Lee;Yun Hu;H. Alexander;S. Libutti;B. Moss;D. Bartlett
我们先前已经证明了全身递送的、复制的牛痘病毒的溶瘤作用。为了增强该载体的肿瘤特异性，我们已经开发了一种组合的胸苷激酶缺失（TK-）和痘苗生长因子缺失（VGF-）的痘苗病毒，并在体外和体内研究其特性。通过同源重组将增强型绿色荧光蛋白（EGFP）的基因插入VGF-牛痘病毒的TK基因座中，产生双缺失突变牛痘病毒（vvDD-GFP）。与来自静息培养物的野生型（WT）、TK-或VGF-病毒相比，用vvDD-GFP感染静息和分裂的NIH 3 T3细胞产生减少的病毒回收，但来自分裂培养物的病毒回收相等。用10（7）空斑形成单位（pfu）的WT、TK-、VGF-或vvDD-GFP牛痘病毒腹膜内注射裸鼠后8天，收获组织和肿瘤用于病毒滴度测定。与其他病毒相比，从注射vvDD-GFP的小鼠脑中未回收到病毒，其他病毒的范围为130至28，000 pfu/mg蛋白质;然而，从肿瘤中回收到等量的病毒。vvDD-GFP没有毒性，因为接受10（8）pfu IP vvDD-GFP的裸鼠存活>100天，而接受WT、VGF-或TK-病毒的小鼠的中位存活期分别仅为6、17和29天。当给予10（9）pfu的vvDD-GFP时，观察到类似的结果。皮下接种裸鼠小鼠结肠腺癌（MC 38）在用10（9）pfu的全身（i. p.）vvDD-GFP与对照相比（注射病毒后12天，平均肿瘤大小为180.71 +/- 35.26 mm（3）vs 2796.79 +/- 573.20 mm（3））。我们的数据表明，TK-和VGF-突变型牛痘病毒在体外静息细胞中显着减毒，并在体内表现出肿瘤特异性复制。由于其增强的安全性、肿瘤选择性和全身递送后的溶瘤作用，它是用于肿瘤定向基因治疗的有前途的载体。
We have demonstrated previously the oncolytic effects of a systemically delivered, replicating vaccinia virus. To enhance the tumor specificity of this vector, we have developed a combined thymidine kinase-deleted (TK-) and vaccinia growth factor-deleted (VGF-) vaccinia virus and investigated its properties in vitro and in vivo. The gene for enhanced green fluorescent protein (EGFP) was inserted into the TK locus of a VGF- vaccinia virus by homologous recombination creating a double-deleted mutant vaccinia virus (vvDD-GFP). Infection of resting and dividing NIH3T3 cells with vvDD-GFP yielded reduced viral recovery compared with wild-type (WT), TK-, or VGF- viruses from resting cultures but equivalent virus recovery from dividing cultures. Eight days after nude mice were injected i.p. with 10(7) plaque-forming units (pfu) of WT, TK-, VGF-, or vvDD-GFP vaccinia virus, tissues and tumor were harvested for viral titer determination. No virus was recovered from the brains of mice injected with vvDD-GFP compared with the other viruses, which ranged from 130 to 28,000 pfu/mg protein; however, equivalent amounts were recovered from tumor. There was no toxicity from vvDD-GFP because nude mice receiving 10(8) pfu of IP vvDD-GFP lived >100 days, whereas mice receiving WT, VGF-, or TK- virus had median survivals of only 6, 17, and 29 days, respectively. Similar results were seen when 10(9) pfu of vvDD-GFP were given. Nude mice bearing s.c. murine colon adenocarcinoma (MC38) had significant tumor regression after treatment with 10(9) pfu of systemic (i.p.) vvDD-GFP compared with control (mean tumor size, 180.71 +/- 35.26 mm(3) versus 2796.79 +/- 573.20 mm(3) 12 days after injection of virus). Our data demonstrate that a TK- and VGF- mutant vaccinia virus is significantly attenuated in resting cells in vitro and demonstrates tumor-specific replication in vivo. It is a promising vector for use in tumor-directed gene therapy, given its enhanced safety profile, tumor selectivity, and the oncolytic effects after systemic delivery.