Establishment of Gene Therapy for Advanced Hepatocellular Carcinoma with Intra- and Extra- Hepatic Metastases

晚期肝细胞癌肝内、肝外转移基因治疗的建立

基本信息

批准号：
10470132
负责人：
KANEKO Shuichi
金额：
$ 4.67万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470132/
关键词：
Gene Therapy Hepatocellular Carcinoma Alpha-fetoprotein Adenovirus

项目摘要

New therapeutic vectors using transcriptional targeting were made for the treatment of advanced hepatocellular carcinoma with intra- and extra-hepatic metastases. Alpha-fetoprotein (AFP) is well known to be specifically expressed in hepatocellular carcinoma. We modified the promoter region of AFP gene by removal of non-functioning parts and enhancing the enhancer sequences, and put the new controlling element of AFP gene into an adenoviral vector. Therapeutic effect and expression analysis of the vectors was tested in transplanted hepatocellular carcinoma in athymic nude mice. Herpes simplex. virus thymidine kinase gene was used as a therapeutic gene.Effect of transcriptional targeting was demonstrated by the AFP controlling element, the anti-tumor effect was low due to the weak promoter activity. Thus, we introduced cre-loxP system into the vector. By the cre-loxP-AFP system, specific and high expression can be obtained in cultured cells and mouse tumors including intra- and extra-hepatic lesions.However, two viruses need to infect simultaneously a cell, and more than 25 MOI are necessary to enable the system. These results indicate large amount of adenovirus should be introduced in a body, which may cause serious problem in future.When HVJ liposome, which is known to have less toxicity, was used with AFP controlling element, repeated administration of the vector confer significant anti-tumor effect in intra-hepatic metastasis model. Transcriptional targeting using AFP promoter is, thus, promising for the treatment of advanced hepatocellular carcinoma with intra- and extra-hepatic metastases.

使用转录靶向的新治疗载体被用于治疗具有肝内和肝外转移的晚期肝细胞癌。众所周知，甲胎蛋白（AFP）在肝细胞癌中特异性表达。我们通过去除非功能部分和增强增强子序列来修饰AFP基因的启动子区域，并将新的AFP基因控制元件放入腺病毒载体中。检测载体对无胸腺裸鼠移植性肝细胞癌的治疗效果和表达分析。单纯疱疹。以病毒胸苷激酶基因作为治疗基因，通过AFP控制元件发挥转录靶向作用，但由于启动子活性较弱，抗肿瘤作用较低。因此，我们将cre-loxP系统引入载体中。通过cre-loxP-AFP系统，可以在培养细胞和小鼠肿瘤（包括肝内和肝外病变）中获得特异性高表达。然而，两种病毒需要同时感染一个细胞，并且需要超过25 MOI才能感染细胞。启用系统。这些结果表明应将大量腺病毒引入体内，这可能会在将来造成严重问题。当已知毒性较小的HVJ脂质体与AFP控制元件一起使用时，重复施用载体可产生显着的抗-肝内转移模型中的肿瘤效应。因此，使用 AFP 启动子的转录靶向有望治疗具有肝内和肝外转移的晚期肝细胞癌。