Selective Cytotoxicity to HER2-Positive Tumor Cells by a Recombinant e23sFv-TD-tBID Protein Containing a Furin Cleavage Sequence
Selective Cytotoxicity to HER2-Positive Tumor Cells by a Recombinant e23sFv-TD-tBID Protein Containing a Furin Cleavage Sequence
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含有弗林蛋白酶裂解序列的重组 e23sFv-TD-tBID 蛋白对 HER2 阳性肿瘤细胞的选择性细胞毒性
DOI：
10.1158/1078-0432.ccr-09-2367
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发表时间：
2010-04
影响因子：
11.5
通讯作者：
药立波
中科院分区：
文献类型：
--
作者：
王芳;Si-Yi Chen;许彦鸣;杨安钢;任静;裘秀春;王立峰;朱青;张英启;宦怡;孟艳玲;药立波
目的：HER 2抗原是乳腺癌细胞上公认的免疫治疗靶点。为了克服传统免疫毒素的免疫原性和全身毒性，我们开发了一种新型的人免疫促凋亡分子，并对其抗肿瘤活性进行了研究。实验设计：重组e23 sFv-TD-tBID由通过10个氨基酸残基的弗林蛋白酶切割序列连接至人活性截短Bid的单链抗HER 2抗体片段组成，其在细菌中表达。测试纯化的e23 sFv-TD-tBID在细胞中的结合、内化和细胞毒性活性，以及在具有已建立的人肿瘤异种移植物的无胸腺裸鼠中的肿瘤定位和抗肿瘤活性。结果：e23 sFv-TD-tBID选择性结合HER 2阳性细胞，并在体外和体内诱导凋亡细胞死亡。对其作用机制的研究表明，e23 sFv-TD-tBID在与HER 2阳性肿瘤细胞表面结合时被内化，经蛋白水解裂解并直接转运至胞质溶胶。将e23 sFv-TD-tBID腹膜内注射到携带人乳腺癌的免疫缺陷小鼠中时，显示出剂量依赖性的抗肿瘤活性。此外，这种免疫促凋亡蛋白，无论是作为单一剂量或与化疗药物组合，显着抑制肿瘤生长，没有任何观察到的毒副作用的小鼠。磁共振成像进一步显示了e23 sFv-TD-tBID在体内对肿瘤的特异性靶向性和良好的穿透性。e23 sFv-TD-tBID对人的治疗价值通过其对原发性患者来源的乳腺肿瘤细胞的细胞毒性作用而不是对内皮细胞的细胞毒性作用来显示。结论：这些数据表明，重组e23 sFv-TD-tBID具有治疗HER 2阳性肿瘤的潜力，并保证进一步测试的临床应用。临床癌症研究; 16（8）; 2284-94。©2010 AACR。
Purpose: The HER2 antigen is a recognized target on breast cancer cells for immunotherapy. To overcome the immunogenicity and systemic toxicity of traditional immunotoxins, a novel human immunoproapoptotic molecule was developed and its antitumor activity was investigated. Experimental Design: Recombinant e23sFv-TD-tBID, consisting of a single-chain anti-HER2 antibody fragment linked to a human active truncated Bid by a 10–amino acid residue furin cleavage sequence, was bacterially expressed. Purified e23sFv-TD-tBID was tested for binding, internalization, and cytotoxic activity in cell and for tumor localization and antitumor activity in athymic nude mice bearing established human tumor xenografts. Results: e23sFv-TD-tBID selectively binds to HER2-positive cells and induces apoptotic cell death in vitro and in vivo. An investigation of its mechanism of action has revealed that e23sFv-TD-tBID was internalized on binding to the surface of HER2-positive tumor cells, proteolytically cleaved and transported directly to cytosol. The antitumor activity of e23sFv-TD-tBID was shown in a dose-dependent manner when injected i.p. into immunodeficient mice bearing human breast carcinomas. Moreover, this immunoproapoptotic protein, either given as a single dose or in combination with chemotherapy agents, significantly inhibited tumor growth without any observed toxic side effects on mice. Magnetic resonance imaging further showed the specific targeting and good penetration of tumors by e23sFv-TD-tBID in vivo. The therapeutic value of e23sFv-TD-tBID to human was shown by its cytotoxic effects on primary patient-derived breast tumor cells but not on endothelial cells. Conclusion: These data suggest that recombinant e23sFv-TD-tBID has therapeutic potential for HER2-positive tumors and warrant further testing for clinical applications. Clin Cancer Res; 16(8); 2284–94. ©2010 AACR.
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DOI：
10.1083/jcb.140.4.733
发表时间：
1998-02-23
期刊：
The Journal of cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
3
作者：
O. Turken;E. Kunter;H. Çermik;T. Ișitmangil;G. Kandemir;M. Yaylacı;A. Ozturk
通讯作者：
O. Turken;E. Kunter;H. Çermik;T. Ișitmangil;G. Kandemir;M. Yaylacı;A. Ozturk
影响因子：
11.2
作者：
Y. Mardor;Yiftach Roth;Z. Lidar;Tali Jonas;Raphael Pfeffer;Stephan E. Maier;M. Faibel;D. Nass;Moshe Hadani;Arie Orenstein;Jack S. Cohen;Zvi Ram
通讯作者：
Y. Mardor;Yiftach Roth;Z. Lidar;Tali Jonas;Raphael Pfeffer;Stephan E. Maier;M. Faibel;D. Nass;Moshe Hadani;Arie Orenstein;Jack S. Cohen;Zvi Ram
DOI：
10.1186/bcr2106
发表时间：
2008
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Grimshaw MJ;Cooper L;Papazisis K;Coleman JA;Bohnenkamp HR;Chiapero-Stanke L;Taylor-Papadimitriou J;Burchell JM
通讯作者：
Burchell JM
DOI：
--
发表时间：
2002-10
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
J. Posey;M. Khazaeli;M. Bookman;A. Nowrouzi;W. Grizzle;J. Thornton;D. Carey;J. Lorenz;A. Sing;C. Siegall;A. Lobuglio;M. Saleh
通讯作者：
J. Posey;M. Khazaeli;M. Bookman;A. Nowrouzi;W. Grizzle;J. Thornton;D. Carey;J. Lorenz;A. Sing;C. Siegall;A. Lobuglio;M. Saleh