Targeted therapy of gastrointestinal cancer using human anti-CEA antibody

人抗CEA抗体靶向治疗胃肠癌

基本信息

批准号：
17591421
负责人：
TANAKA Toshihiro
金额：
$ 2.24万
依托单位：
Fukuoka University (2006)Sapporo Medical University (2005)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591421/
关键词：
adenovirus CEA antibody gastric cancer gene therapy

项目摘要

We constructed a fiber modified Ad5 vector (Adv-FZ33) in which an IgG-binding domain (Z33) derived from staphylococcal protein A was inserted into the HI loop of knob protein. We evaluated on both in vitro and in vivo levels the extent of retargeting towards and therapeutic effectiveness against CEA-positive gastric cancers when using the anti-CEA antibody complex with this modified vector. In vitro LacZ gene expression after infection with Adv-FZ33 vectors conjugated to anti-CEA mAb was approximately 20 times higher than that obtained with vector conjugated to the control mAb. We generated Ax3CAUP-FZ33, which is an Adv-FZ33 derivative vector expressing a therapeutic gene, namely E. coli uracil phosphoribosyltransferase (UPRT) which converts 5FU directly to 5-fluorouridine monophosphate. When conjugated with the anti-CEA mAb, Ax3CAUP-FZ33 enhanced the cytotoxicity of 5FU (19.7-fold in terms of IC_<50> values) against gastric cancer cells. Nextl, we examined the survival effects of Ax3U … More P-FZ33 conjugated with anti-CEA mAb plus 5FU in mice with peritoneal disseminated gastric cancer. The median survival time of the Ax3CAUP-FZ33/anti-CEA mAb/5FU group was significantly longer than those of the PBS and 5FU only treatment groups (p<0.01, versus PBS and 5FU only groups).Furthermore, we examined which subtype of IgG was effective on gene delivery mediated by Adv-FZ33 and anti-CEA mAb. BIACORE analysis showed that human IgG1 and IgG4 had 1,000-fold higher affinity constants (Ka) for protein A compared with mouse IgG1, whereas only 10-fold higher than mouse IgG2a or IgG3. In CEA-expressing cells, the transgene expression using Adv-FZ33 and any subtype of anti-CEA human IgG was significantly increased compared with combinations of any type of virus-anti-CEA mouse IgG. Especially, human IgG4 mediated transduction efficiency showed 200-fold enhancements compared with mouse IgG1. We found that the reactivity of Ab and protein A played a key role in the Ab dependent gene delivery by Adv-FZ33. Less

我们构建了一种纤维修饰的 Ad5 载体 (Adv-FZ33)，其中将源自葡萄球菌蛋白 A 的 IgG 结合域 (Z33) 插入到旋钮蛋白的 HI 环中，我们在体外和体内水平评估了这种作用的程度。使用抗 CEA 抗体复合物与该修饰载体结合的 Adv-FZ33 载体感染后的体外 LacZ 基因表达，对 CEA 阳性胃癌进行重新靶向和治疗效果。抗 CEA mAb 的结果比与对照 mAb 缀合的载体获得的结果高约 20 倍。我们生成了 Ax3CAUP-FZ33，它是表达治疗基因（即大肠杆菌尿嘧啶磷酸核糖基转移酶 (UPRT)）的 Adv-FZ33 衍生载体。 5FU 直接与 5-氟尿苷单磷酸结合当与抗 CEA mAb 结合时，Ax3CAUP-FZ33 得到增强。 5FU（IC_<50> 值的 19.7 倍）对胃癌细胞的细胞毒性接下来，我们检查了 Ax3U … 更多 P-FZ33 与抗 CEA 单克隆抗体加 5FU 结合对腹膜播散性胃癌细胞的生存影响。 Ax3CAUP-FZ33/抗-CEA mAb/5FU 组的中位生存时间显着长于 PBS 和 PBS 组。仅 5FU 治疗组（与仅 PBS 和 5FU 组相比，p<0.01）。此外，我们检查了哪种 IgG 亚型对 Adv-FZ33 介导的基因递送有效，抗 CEA mAb 分析显示人 IgG1 和 IgG4 具有特异性。与小鼠 IgG1 相比，蛋白 A 的亲和常数 (Ka) 高 1,000 倍，而仅比小鼠 IgG2a 或 10 倍高在表达CEA的细胞中，与任何类型的病毒-抗CEA小鼠IgG的组合相比，使用Adv-FZ33和任何亚型的抗CEA人IgG的转基因表达显着增加，尤其是人IgG4介导的转导效率。与小鼠 IgG1 相比，我们发现 Ab 和 Protein A 的反应性在 Adv-FZ33 的 Ab 依赖性基因传递中发挥了关键作用。