Targeted therapy of gastrointestinal cancer using human anti-CEA antibody

人抗CEA抗体靶向治疗胃肠癌

• 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

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

我们构建了一个修饰的AD5载体（ADV-FZ33），其中将源自葡萄球菌蛋白A衍生的IgG结合域（Z33）插入旋钮蛋白的HI环中。我们在体外和体内水平上都评估了对CEA阳性胃癌的重新定位和治疗有效性的程度，当时使用抗CEA抗体复合物与该修饰的载体。与抗CEA MAB相连的Adv-FZ33载体感染后的体外LACZ基因表达，是与对照mAb相连的载体获得的20倍。我们生成了AX3CAUP-FZ33，它是一种表达治疗基因的Adv-FZ33衍生物载体，即大肠杆菌尿嘧啶磷酸糖基转移酶（UPRT），可直接将5FU转换为5-氟尿苷单磷酸盐。当与抗CEA MAB结合时，AX3CAUP-FZ33对胃癌细胞增强了5FU（IC_ <50>值的19.7倍）的细胞毒性。 NextL，我们检查了AX3U的生存效应…更多的P-FZ33与腹膜传播胃癌的小鼠中的抗CEA MAB加5FU结合了5FU。 AX3CAUP-FZ33/anti-CEA MAB/5FU组的中位生存时间明显比PBS和仅5FU治疗组的中位生存时间（P <0.01，与PBS和仅5FU组）。FURTHERMORE。我们检查了IgG的igG子类型对Gene的递送有效介导了Gene的递送，该子型对ADV-FZ33和Antigi-CEA介导了Gene递送。 BIACORE分析表明，与小鼠IgG1相比，人IgG1和IgG4的亲和力常数（Ka）高1,000倍，而仅比小鼠IgG2A或IgG3高10倍。在表达CEA的细胞中，与任何类型的病毒-Anti-CEA小鼠IgG的组合相比，使用ADV-FZ33和任何抗CEA人IgG的任何亚型的转基因表达显着增加。尤其是，与小鼠IgG1相比，人IgG4介导的翻译效率显示了200倍。我们发现，AB和蛋白A的反应性在ADV-FZ33中递送AB依赖基因中起关键作用。较少的

项目成果

Re-targeting of cytotoxic T lymphocytes and/or natural killer cells to CEA-expressing tumor cells with anti-CEA antibody activity.

将细胞毒性 T 淋巴细胞和/或自然杀伤细胞重新靶向具有抗 CEA 抗体活性的表达 CEA 的肿瘤细胞。

• 发表时间: 2005
• 期刊: Anticancer Res. 25(6)
• 作者: Kuroki;Ma.;Hachimine;K.;Huang J.;Shibaguchi;H.;Kinugasa;T.;Maekawa;S.;Kuroki;Mo.
• 通讯作者: Mo.

Recent Development in Gene Therapy

基因治疗的最新进展

• 发表时间: 2007
• 作者: Tanaka T;Hamada H;Kuroki Ma;et al.
• 通讯作者: et al.

Targeting of cancer gene therapy with antibodies or their genes against tumor-associated antigens.

使用抗肿瘤相关抗原的抗体或其基因来靶向癌症基因治疗。

• 发表时间: 2005
• 期刊: Gene Ther.Mol.Biol. 9
• 作者: 田中純子;片山恵子;Kuroki M.
• 通讯作者: Kuroki M.

Gene transfer of the CD40-ligand to human dendritic cells induces NK-mediated antitumor effects against human carcinoma cells.

CD40 配体基因转移至人树突状细胞可诱导 NK 介导的针对人癌细胞的抗肿瘤作用。

• 发表时间: 2007
• 期刊: Int. J. Cancer 120(7)
• 作者: Tomihara K;Kato K;Masuta Y;Nakamura K;Tanaka T;Hiratsuka H;Hamada H
• 通讯作者: Hamada H

Cancer Immunotherapy with Chimeric Immune Receptors (CIRs) : A Focus on Their Antitumor Activity.

嵌合免疫受体 (CIR) 的癌症免疫疗法：关注其抗肿瘤活性。

• 发表时间: 2006
• 期刊: Med. Bull. Fukuoka Univ. 33(3)
• 作者: Shibaguchi H;Tanaka T;Kuroki Ma;et al.
• 通讯作者: et al.