Gene therapy for intractable gastrointestinal tumors with tumor promoter-mediated oncolyitc adenoviruses

肿瘤启动子介导的溶瘤腺病毒对难治性胃肠道肿瘤的基因治疗

• 批准号: 15590706
• 负责人: KAWAMURA Kiyoko
• 金额: $ 2.24万
• 依托单位: Chiba Cancer Center Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590706/
• 关键词: Gene Therapy; Tumor promoter; Transcription regulatory region; Oncolytic virus; Midkine; Survivin; E1A; SCID mouse

We have constructed adenoviruses in which the E1A transcript was regulated with a 604 bp-regulatory region of the midkine gene (Ad/MK) ; the transcription of the gene is upregulated in a number of human gastrointestinal tumors but is restricted in normal adult tissues. Ad/MK induced greater E1A expression than wild-type Ad (Ad/WT) in human fibroblasts immortalized by the expression of dominant-negative p53 gene, but not in the parent normal fibroblasts. Ad/MK were cytotoxic to the immortalized fibroblasts compared with Ad/WT accordingly. Human hepatocellular carcinoma(HCC) cells were 500 times more susceptible to Ad/MK than normal fibroblasts, and replication of Ad/MK was 1000 times greater than that of Ad/WT in the HCC cells. Intratumoral injection of Ad/MK but not replication-defective Ad suppressed subsequent growth of solid HCC tumors developed in SCID mice. Histological examinations revealed massive necrosis of the tumors injected with Ad/MK. These data suggested that Ad/MK are oncolytic Ad that preferentially destroy tumors. We also examined the feasibility of oncolytic Ad with different a tumor promoter. Expression of the survivin gene is regulated with the cell cycle and is upregulated in tumors ; thereby the regulatory region is a candidate to control E1A expression. We identified a 500-bp region that mediated the preferential expression in tumors and is currently preparing oncolytic Ad with the promoter.

我们构建了腺病毒，其中 E1A 转录本受中期因子基因 (Ad/MK) 的 604 bp 调控区的调控；该基因的转录在许多人类胃肠道肿瘤中上调，但在正常成人组织中受到限制。在通过显性失活 p53 基因表达永生化的人成纤维细胞中，Ad/MK 诱导比野生型 Ad (Ad/WT) 更高的 E1A 表达，但在亲本正常成纤维细胞中则不然。相应地，与Ad/WT相比，Ad/MK对永生化成纤维细胞具有细胞毒性。人肝细胞癌细胞（HCC）对Ad/MK的敏感性比正常成纤维细胞高500倍，并且HCC细胞中Ad/MK的复制比Ad/WT高1000倍。瘤内注射 Ad/MK 但不抑制复制缺陷型 Ad 抑制 SCID 小鼠中实体 HCC 肿瘤的后续生长。组织学检查显示注射 Ad/MK 的肿瘤出现大量坏死。这些数据表明Ad/MK是优先破坏肿瘤的溶瘤Ad。我们还研究了具有不同肿瘤启动子的溶瘤Ad的可行性。生存素基因的表达受细胞周期调节，在肿瘤中表达上调；因此该调控区是控制E1A表达的候选区域。我们鉴定了一个介导肿瘤中优先表达的 500 bp 区域，目前正在用启动子制备溶瘤 Ad。

项目成果

Saisho H, Tagawa M, et al.: "Down-regulated expression of CCAAT/enhancer binding protein α and β genes in human hepatocellular carcinoma : a possible prognostic marker."Anticancer Res.. 23. 351-354 (2003)

Saisho H、Takawa M 等人：“人肝细胞癌中 CCAAT/增强子结合蛋白 α 和 β 基因的下调表达：一个可能的预后标记。”Anticancer Res.. 23. 351-354 (2003)

Kawamura K, Saisho H, Tagawa M, et al.: "Expression of the interleukin-21 gene in murine colon carcinoma cells generates systemic immunity in the inoculated hosts."Cancer Gene Ther. 10. 187-192 (2003)

Kawamura K、Saisho H、Takawa M 等人：“小鼠结肠癌细胞中白细胞介素 21 基因的表达在接种的宿主中产生全身免疫。”癌症基因疗法。

Kawamura K, Tagawa M, et al.: "Fas ligand-expressing tumors induce tumor-specific protective immunity in the inoculated hosts but vaccination with the apoptotic tumors suppresses antitumor immunity."Cancer Gene Ther. 10. 134-140 (2003)

Kawamura K、Takawa M 等人：“表达 Fas 配体的肿瘤在接种的宿主中诱导肿瘤特异性保护性免疫，但接种凋亡肿瘤会抑制抗肿瘤免疫。”Cancer Gene Ther。

Regulatory regions of growth-related genes can activate an exogenous gene of the α-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells.

生长相关基因的调节区域可以激活甲胎蛋白启动子的外源基因，其程度与人肝细胞癌细胞中的水平相当。

• 发表时间: 2003
• 期刊: Anticancer Res. 23
• 作者: Tomizawa;M.;Saisho;H.;Tagawa;M.
• 通讯作者: M.

Fas ligand-expressing tumors induce tumor-specific protective mmunity in the inoculated hosts but vaccination with the apoptotic tumors suppresses antitumor immunity.

表达 Fas 配体的肿瘤在接种的宿主中诱导肿瘤特异性保护性免疫，但用凋亡肿瘤进行疫苗接种会抑制抗肿瘤免疫。

• 发表时间: 2003
• 期刊: Cancer Gene Ther. 10
• 作者: Tada;Y.;O-Wang;J.;Wada;A.;Takiguchi;Y.;Tatsumi;K.;Kuriyama;T.;Sakiyama;S.;Tagawa;M.
• 通讯作者: M.