Oncolytic adenovirus modified to increase its infectivity for gastrointestinal cancer

溶瘤腺病毒经过修饰以增加其对胃肠道癌症的感染性

• 批准号: 16591381
• 负责人: TAGAWA Masatoshi
• 金额: $ 2.3万
• 依托单位: Chiba Cancer 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: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591381/
• 关键词: Gastrointestival tumors; Gene therapy; Transcriptional control region; E1A; E1B region; Oncolytic adenovirus; Fiber region; Suvivin; Shuttle vector; 腫瘍プロモーター; 腫瘍融解性ウイルス; ファイバー・ノブ構造; サバイビン; ミッドカイン

Adenovirus (Ad) has a strong cytotoxic activity and the E1A protein, one of the early transcriptional products, has a pivotal role in viral replication in the infected cells. Ad modified to control the E1A gene expression with a putative tumor promoter can thereby be oncolytic. We examined the regulatory regions of the genes which were predominantly expressed in tumors with the luciferase assay and found that a 500 bp and a 600 bp upstream regions of the survivin and midkine genes, respectively, were responsible for the transcriptional control. The cellular receptors of the Ad type 5 is CAR (Coxsackievirus and Adenovirus Receptor) and the expression is often downregulated in gastrointestinal tumors. In contrast, the expression of CD46, which is a receptor of Ad type 35, is rather upregulated in the tumors and consequently, the Ad bearing CD46 binding sites can increase the infectivity to the tumors. We thereby constructed a vector system consisting of vector DNA that could activate the E1A and E1B genes with an exogenous regulatory region and another vector that contained the rest of the whole Ad sequences, in which the fiber-knob region was replaced with the Ad type 35-derived one. In vitro ligation of the vectors enables the construction of the Ad bearing the type 35 fiber-knob structure in the E3 region. The oncolytic Ad with the type 35 fiber-knob can be produced with easy compared with conventional methods that required in vivo recombination. The chimeric oncolytic Ad prepared with our vector system was more cytotoxic to CAR-low human tumors than conventional type 5 oncolytic Ad and was as cytotoxic as the type 5 Ad to CAR-high tumors.

腺病毒（AD）具有强大的细胞毒性活性，E1A蛋白（早期转录产物之一）在感染细胞中的病毒复制中具有关键作用。 AD修饰以用推定的肿瘤启动子来控制E1a基因表达，从而有溶瘤。我们检查了主要在荧光素酶测定中以肿瘤表达的基因的调节区域，发现分别为Survivin和Midkine基因的500 bp和600 bp的上游区域负责转录对照。 AD 5型的细胞受体是CAR（Coxsackievivirus和腺病毒受体），并且在胃肠道肿瘤中通常会下调表达。相反，CD46的表达是AD 35型的受体，在肿瘤中相当上调，因此，带有CD46结合位点的AD可以增加对肿瘤的感染性。因此，我们构建了一个由矢量DNA组成的矢量系统，该矢量系统可以激活具有外源调节区域的E1A和E1B基因，而另一个包含整个AD序列的载体，其中纤维旋钮区域被AD AD型35型替换为35级。向量的体外连接能够构建E3区域中35型纤维旋钮结构的广告。与需要体内重组的常规方法相比，具有35型纤维旋钮的溶瘤AD可以轻松产生。与传统的5型溶瘤AD相比，使用我们的矢量系统制​​备的嵌合肿瘤AD对型号的人类肿瘤具有更大的细胞毒性，并且与5型AD对汽车高肿瘤一样的细胞毒性。

项目成果

Vaccination of apoptotic Fas ligand-expressing tumors decreased antitumor responses by enhanced production of immunosuppressive cytokines.

表达凋亡 Fas 配体的肿瘤的疫苗接种可通过增强免疫抑制细胞因子的产生来降低抗肿瘤反应。

• 发表时间: 2005
• 期刊: Anticancer Res. 25
• 作者: Wada A;Tagawa M et al.
• 通讯作者: Tagawa M et al.

Elevated expression of DNA polymerase x expression in human lung cancer is associated with p53 inactivation: Negative regulation of POLKpromoter activity by p53.

人肺癌中 DNA 聚合酶 x 表达的升高与 p53 失活相关：p53 对 POLK 启动子活性的负调节。

• 发表时间: 2004
• 期刊: Int.J.Oncol. 25
• 作者: Wang;Y.Q.;Seimiya;M.;Kawamura;K.;Yu;L.;Ogi;T.;Takenaga;K.;Shishikura;T.;Nakagawara;A.;Sakiyama;S.;Tagawa;M.;O-Wang;J.
• 通讯作者: J.

Increased infectivity of adenovirus type 5 bearing type 11 or type 35 fibers to human esophageal and oral carcinoma cells.

• DOI: 10.3892/or.14.4.831
• 发表时间: 2005-10
• 期刊: Oncology reports
• 影响因子: 4.2
• 作者: Ling Yu;H. Takenobu;O. Shimozato;K. Kawamura;Y. Nimura;N. Seki;K. Uzawa;H. Tanzawa;H. Shimada;T. Ochiai;M. Tagawa

Expression of the fetal-oncogenic fibroblast growth factor-8/17/18 subfamily in human hematopoietic tumors.

胎儿致癌成纤维细胞生长因子 8/17/18 亚家族在人类造血肿瘤中的表达。

• 发表时间: 2005
• 期刊: Biochem Biophys Res Commun 335
• 作者: Nezu;M.
• 通讯作者: M.

Midkine promoter-driven suicide gene expression and -mediated adenovirus replication produced cytotoxic effects immortalized and tumour cells.

中期因子启动子驱动的自杀基因表达和介导的腺病毒复制产生永生化和肿瘤细胞的细胞毒性作用。

• 发表时间: 2004
• 期刊: European Journal of Cancer 40
• 作者: Chiyo;M.;Shimozato;O.;Iizasa;T.;Fujisawa;T.;Tagawa;M.;Yu Ling;Kawamura Kiyoko;Yu Ling
• 通讯作者: Yu Ling