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Transoriptional Tayiting of herpes simplex virus for cell-specific replication

单纯疱疹病毒的转录Tayiting用于细胞特异性复制

基本信息

批准号：
07671513
负责人：
MIYATAKE Shin-Ichi
金额：
$ 1.6万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671513/
关键词：
herpes simplex virus gene therapy albumin hepatocellular coraimoma gene promotor

项目摘要

We have developed a strategy for viral-mediated tumor therapy, where herpes simplex virus (HSV) replication and associated cytotoxicity are limited to a specific cell-type by the regulated expression of an essential immediate-early gene. A HSV vector, G92A,was constructed whose growth is restricted to albumin-expressing, dividing cells. G92A contains an albumin enhancer/promoter-ICP4 transgene within the thymidine kinase (tk) gene of the HSV-1 ICP4 deletion mutant d120. In addition, it contains the E.coli LacZ gene upstream of the ICP4 transgene, under control of the tk promoter. Therefore, G92A plaques stain bule with X-gal (lacZ+) in the presence of gancilovir (tk-). In Vitro, G92A replicates well in 3 human hepatoma (albumin-expressing) cell lines, destroying them, and not in 8 non-albumin-expressing human tumor cell lines. This cell-specificity was conserved in vivo, where the growth of established subcutaeous hepatoma tumors was significantly inhibited by a single inoculation of G92A,whereas, there was no effect on the growth of non-albumin expressing tumors. Mice inoculated intrahepatically with G92A,at doese that were 100% lethal with wild-type HSV,exhibited no symptoms of disease. These studies demonstrate the feasibility of confining a productive, lytic infection to defined cell types. HSV vectors containing other cell-specific regulatory regions should confer targeting to a variety of tumor types.

我们已经开发了一种病毒介导的肿瘤治疗策略，其中单纯疱疹病毒（HSV）的复制和相关的细胞毒性被限制在特定的细胞类型中，通过调节一个必要的即时早期基因的表达。构建了HSV载体G92A，其生长仅限于表达白蛋白的分裂细胞。G92A在HSV-1 ICP4缺失突变体d120的胸苷激酶（tk）基因中含有白蛋白增强子/启动子ICP4转基因。此外，它含有ICP4转基因上游的大肠杆菌LacZ基因，受tk启动子控制。因此，G92A斑块在更西洛韦（tk-）存在下用X-gal （lacZ+）染成蓝色。在体外，G92A在3个表达白蛋白的人肝癌细胞系中复制良好，破坏了它们，而在8个不表达白蛋白的人肿瘤细胞系中不复制。这种细胞特异性在体内是保守的，单次接种G92A可以显著抑制已建立的皮下肝癌肿瘤的生长，而对不表达白蛋白的肿瘤的生长没有影响。小鼠肝内接种G92A，剂量为野生型HSV的100%致死，没有表现出疾病症状。这些研究证明了将一种多产的、溶解性的感染限制在特定的细胞类型上的可行性。含有其他细胞特异性调控区域的HSV载体应赋予多种肿瘤类型的靶向性。