Gene Therapy for analignant using fumor-Specific HSV

载体的组织或细胞特异性靶向对于基因治疗的成功至关重要。我们描述了一种新的病毒介导的基因治疗方法，其中病毒复制和相关的细胞毒性被限制到一个特定的细胞类型的一个重要的立即病毒基因产物的表达受到管制。这是说明与单纯疱疹病毒1型载体（G92 A），其生长仅限于白蛋白表达细胞。通过将白蛋白增强子/启动子-ICP 4转基因插入突变型单纯疱疹病毒1型dl 20的胸苷激酶基因中构建G92 A，缺失ICP 4基因的两个拷贝。该载体还含有在胸苷激酶启动子（一种病毒早期启动子）控制下的大肠杆菌lacZ基因，以允许容易地检测含有复制载体的感染细胞。在成人中，白蛋白仅在肝脏和肝细胞癌中表达，并受到转录调控。G92 A在两个表达白蛋白的人肝癌细胞系中有效复制，但在三个不表达白蛋白的人非肝癌肿瘤细胞系中无效，而所有细胞系对组织非特异性HSV重组体hrR 3同样敏感。然而，尽管我们尝试了多次同源重组，我们仍然没有获得目标HSV。同时，我们也将重组HSV，hrR 3用于血管病变，如球囊血管成形术后再狭窄。目前我们正在构建血管平滑肌细胞特异性复制型HSV。我们还构建了表达bFGF的复制缺陷型腺病毒载体，用于体内神经元保护和血管生成。

Tissue- or cell-specific targeting of vectors is critical to the success of gene therapy. We describe a novel approach to viral-mediated gene therapy, where viral replication and associated cytotoxicity are limited to a specific cell-type by the regulated expression of an essential immediately viral gene product. This is illustrated with a herpes simples virus type 1 vector(G92A)whose growth is restricted to albumin-expressing cells. G92A was constructed by inserting an albumin enhancer/promoter-ICP4 transgene into the thymidine kinase gene of mutant herpes simples virus type 1 dl20, deleted for both copies of the ICP4 gene. This vector also contains the Escherichia coli lacZ gene under control of the thymidine kinase promoter, a viral early promoter, to permit easy detection of infected cells containing replicating vector. In the adult, albumin is expressed uniquely in the liver and in hepatocellular carcinoma and is transcriptionally regulated. G92A efficiently replicated in vitro in two human hepatoma cell lines expressing albumin, but not in three human nonhepatoma, albumin-non-expressing tumor cell lines, while all cell lines were equally susceptible to a tissue ono-specific HSV recombinant, hrR3.We applied the same strategy for the construction of glioma-specific replication competent HSV using GFAP promoter and enhancer units. However, although we tried homologous recombination several times, we have not yet obtained the aimed HSV.While, we also applied the recombinant HSV, hrR3 for vascular lesions such as re-stenosis after balloon angioplasty. Now we are constructing vascular smooth muscle cell-specific replication competent HSV. We also constructed replication-incompetent adenovirus vectors expressing bFGF for the purpose of neuronal protection and angiogenesis in vivo.