Gene therapy for hepatocellular carcinoma using EBV plasmid vector

EBV质粒载体对肝细胞癌的基因治疗

• 批准号: 11670523
• 负责人: IWAI Masaki
• 金额: $ 2.18万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670523/
• 关键词: Hepatocellular carcinoma; Gene therapy; EBV plasmid vector; Nonviral vector; α-Fetoprotein; 非ウィルスベクター; α-Fctoprotein

We investigated plasmid vector containing EBNA1/oriP and inserted gene of Herpes simplex virus 1-Thymidine kinase (HSV 1-Tk) into the vector. We transduced the vector into cell line of human hepatoma and Ewing's sarcoma with cationic liposome or polyamidoamine dendrimer, and suicidal effect of HSV1-TK driven by EBNA1/oriP was found to be in 3 to 10 hundreds times as strong as by control plasmid driven by general promotor under administration of gancyclovir (Y Harada, M Iwai, et al. Cancer Gene Ther 7 ; 27, 2000. H Maruyama-Tabata, et al. Gene Ther 7 ; 53, 2000).We transduced vector containing EBNA1/oriP with CEA promotor into CEA-positive cell line of cholangiocarcinoma (CCC) by polyamidoamine dendrimer and suicidal effect was found to be specific for CEA-positive CCC (S Tanaka, M Iwai et al. Cancer Gene Ther 7 ; 1241, 2000). Then, we reported that suicide gene therapy with EBNA1/oriP system was effective in vitro and in vivo, and specific effect for cancer was gained by using tumor promotor in EBNA1/oriP system. In addition we studied structure and function of gap junction in ontogenic liver to clarify mechanism of "Bystander effect" in gene therapy (M Iwai, et al. J Hepatol 32 ; 11, 2000). Judging from our investigation, we should apply EBNA1/oriP system containing AFP promotor as well as gene for gap junctional protein for suicide gene therapy ofAFP-positive hepatoma cells.

我们研究了含EBNA 1/oriP的质粒载体，并将单纯疱疹病毒1型胸苷激酶（HSV 1-Tk）基因插入到该载体中。我们用阳离子脂质体或聚酰胺胺树枝状聚合物将载体转导到人肝癌和尤文氏肉瘤的细胞系中，并且发现在施用更昔洛韦的情况下，由EBNA 1/oriP驱动的HSV 1-TK的自杀效应是由通用启动子驱动的对照质粒的3至1000倍（Y Harada，M Iwai，et al. Cancer Gene Ther 7 ; 27，2000. H Maruyama-Tabata等，Gene Ther 7 ; 53，2000）.我们通过聚酰胺胺树状聚合物将含有带有CEA启动子的EBNA 1/oriP的载体转导到胆管癌（CCC）的CEA阳性细胞系中，发现自杀效应对CEA阳性CCC是特异性的（S Tanaka，M Iwai等，Cancer Gene Ther 7 ; 1241，2000）。在此基础上，我们报道了EBNA 1/oriP系统的自杀基因治疗在体内外均有效，并在EBNA 1/oriP系统中加入肿瘤启动子，获得了特异性的肿瘤治疗效果。此外，我们研究了个体发育肝脏中间隙连接的结构和功能，以阐明基因治疗中“旁观者效应”的机制（M Iwai，et al. J Hepatol 32 ; 11，2000）。因此，我们认为有必要将含AFP启动子和缝隙连接蛋白基因的EBNA 1/oriP系统用于AFP阳性肝癌细胞的自杀基因治疗。

项目成果

H Maruyama-Tabata, Y Harada, T Matsumura, E Satoh, F Cui, M Iwai, M Kita, S Hibi, J Imanishi, T Sawada, O Mazda: "Effective suicide gene therapy in vivo by EBV-based plasmid vector coupled with polyamidoamine dendrimer."Gene Ther. 7. 53-60 (2000)

H Maruyama-Tabata、Y Harada、T Matsumura、E Satoh、F Cui、M Iwai、M Kita、S Hibi、J Imanishi、T Sawada、O Mazda：“通过基于 EBV 的质粒载体与

M Iwai et al.: "Cholestatic ** in two patients with primary amyloidosis-Ultrastructural tihadiys〜"J Clin Gastroenteral. 28(2). 162-166 (1999)

M Iwai 等人：“两名原发性淀粉样变性患者的胆汁淤积**-超微结构 tihadiys〜”J Clin Gastroenteral 28（2）162-166（1999）。

Iwai M,Morikawa T, et al.: "Biological significance of AFP expression in liver injury induced by CC14"Acta histochem cytochem. 33. 17-22 (2000)

Iwai M、Morikawa T 等人：“CC14 诱导的肝损伤中 AFP 表达的生物学意义”组织化学细胞化学学报。

Y Harada,H Maruyama-Tabata,E Satoh,M Iwai, et al.: "Superfect reagent-mediated transfection with on EBV-based plasmid vector"Qiagen News. 5. 12-14 (2000)

Y Harada、H Maruyama-Tabata、E Satoh、M Iwai 等人：“Superfect 试剂介导的基于 EBV 的质粒载体的转染”Qiagen 新闻。

M Iwai, T Morikawa, A Muramatsu, S Tanaka, T Mori, Y Harada, T Okanoue, K Kashima., M Ishii: "Biological significance of AFP expression in liver injury induced by CCl_4."Acta histochem cytochem. 33. 17-22 (2000)

M Iwai、T Morikawa、A Muramatsu、S Tanaka、T Mori、Y Harada、T Okanoue、K Kashima.、M Ishii：“AFP 表达在 CCl_4 诱导的肝损伤中的生物学意义。”组织化学细胞化学学报。