Recombinant anti-podoplanin (NZ-1) immunotoxin for the treatment of malignant brain tumors.
Recombinant anti-podoplanin (NZ-1) immunotoxin for the treatment of malignant brain tumors.
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DOI：
10.1002/ijc.27919
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发表时间：
2013-05-15
影响因子：
6.4
通讯作者：
Bigner, Darell D.
中科院分区：
文献类型：
--
作者：
Chandramohan, Vidyalakshmi;Bao, Xuhui;Kaneko, Mika Kato;Kato, Yukinari;Keir, Stephen T.;Szafranski, Scott E.;Kuan, Chien-Tsun;Pastan, Ira H.;Bigner, Darell D.
关键词：
目前的研究表明，胶质母细胞瘤(D2159 MG、D08-0308 MG和D08-0493 MG)和髓母细胞瘤(D283MED、D425MED和DAOY)异种移植瘤和细胞系中都存在非常高水平的胶质瘤抗原泊多拉宁(>；90%)。我们从NZ-1杂交瘤细胞中构建了针对泊多拉宁的新型重组单链抗体可变区片段(ScFv)NZ-1。将NZ-1-scFv与假单胞菌外毒素A融合，携带一个KDEL多肽(NZ-1-PE38KDEL)。重链可变区和轻链可变区之间的二硫键进一步稳定了免疫毒素，构建了NZ-1-(ScdsFv)-PE38KDEL。NZ-1-(ScdsFv)-PE38KDEL对胶质母细胞瘤和髓母细胞瘤细胞具有明显的反应性。NZ-1-(ScdsFv)、NZ-1-(ScdsFv)-PE38KDEL和NZ-1抗体对PodoPlanin多肽的亲和力分别为2.1×10−8M、8.0×10−8M和3.9×10−10M。在蛋白质稳定性实验中，NZ-1-(ScdsFv)-PE38KDEL在37℃下孵育3d后，其活性保持了33-98%，而NZ-1-PE38KDEL的活性下降到初始水平的13%。体外测定了NZ-1-(ScdsFv)-PE38KDEL对人胶质母细胞瘤D2159 MG、D08-0308 MG、D08-0493 MG以及髓母细胞瘤D283MED、D425MED和DOAY异种移植瘤及细胞系的细胞毒作用。NZ-1-(ScdsFv)-PE38KDEL免疫毒素具有较强的细胞毒性，其IC50为1.6~29 ng/mL。值得注意的是，NZ-1-(ScdsFv)-PE38KDEL在D2159 MG和D283MED体内肿瘤模型中显示出肿瘤生长延迟，平均分别为24天(P<；0.001)和21天(P<；0.001)。关键是，在D425MED颅内肿瘤模型中，NZ-1-(ScdsFv)-PE38KDEL使生存率增加了41%(P≤0.001)。在临床前研究中，NZ-1-(ScdsFv)-PE38KDEL显示出作为恶性脑肿瘤靶向性药物的巨大潜力。
Current study demonstrates the glioma tumor antigen podoplanin to be present at very high levels (>90%) in both glioblastoma (D2159MG, D08-0308MG, and D08-0493MG) and medulloblastoma (D283MED, D425MED, and DAOY) xenografts and cell line. We constructed a novel recombinant single-chain antibody variable region fragment (scFv), NZ-1, specific for podoplanin from the NZ-1 hybridoma. NZ-1-scFv was then fused to Pseudomonas exotoxin A, carrying a C-terminal KDEL peptide (NZ-1-PE38KDEL). The immunotoxin was further stabilized by a disulfide (ds) bond between the heavy-chain and light-chain variable regions as the construct NZ-1-(scdsFv)-PE38KDEL. NZ-1-(scdsFv)-PE38KDEL exhibited significant reactivity to glioblastoma and medulloblastoma cells. The affinity of NZ-1-(scdsFv), NZ-1-(scdsFv)-PE38KDEL and NZ-1 antibody, for podoplanin peptide was 2.1×10−8 M, 8.0×10−8 M, and 3.9×10−10 M, respectively. In a protein stability assay, NZ-1-(scdsFv)-PE38KDEL retained 33-98% of its activity while that of NZ-1-PE38KDEL declined to 13% of its initial levels after incubation at 37°C for 3 days. In vitro cytotoxicity of the NZ-1-(scdsFv)-PE38KDEL was measured in cells isolated from glioblastoma xenografts, D2159MG, D08-0308MG, D08-0493MG, and in the medulloblastoma D283MED, D425MED, and DOAY xenografts and cell line. The NZ-1-(scdsFv)-PE38KDEL immunotoxin was highly cytotoxic, with an IC50 in the range of 1.6–29 ng/mL. Significantly, NZ-1-(scdsFv)-PE38KDEL demonstrated tumor-growth delay, averaging 24 days (P<0.001) and 21 days (P<0.001) in D2159MG and D283MED in vivo tumor models, respectively. Crucially, in the D425MED intracranial tumor model, NZ-1-(scdsFv)-PE38KDEL caused a 41% increase in survival (P≤0.001). In preclinical studies, NZ-1-(scdsFv)-PE38KDEL exhibited significant potential as a targeting agent for malignant brain tumors.
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影响因子：
158.5
作者：
Stupp, R;Mason, WP;Ryan, G
通讯作者：
Ryan, G
影响因子：
3.1
作者：
Kato Y;Vaidyanathan G;Kaneko MK;Mishima K;Srivastava N;Chandramohan V;Pegram C;Keir ST;Kuan CT;Bigner DD;Zalutsky MR
通讯作者：
Zalutsky MR
影响因子：
4.2
作者：
Li YM;Hall WA
通讯作者：
Hall WA
DOI：
10.1016/j.bbrc.2006.08.171
发表时间：
2006-11-03
影响因子：
3.1
作者：
Kato, Yukinari;Kaneko, Mika Kato;Osawa, Motoki
通讯作者：
Osawa, Motoki
影响因子：
6
作者：
Kunita, Akiko;Kashima, Takeshi G.;Fujita, Naoya
通讯作者：
Fujita, Naoya