SPECT imaging of neuropilin receptor type-1 expression with 131I-labeled monoclonal antibody.
As a novel co-receptor for vascular endothelial growth factor (VEGF), neuropilin receptor type-1 (NRP-1) is overexpressed in several cancers and metastases, and serves as an attractive target for cancer molecular imaging and therapy. Previous single photon emission computerized tomography (SPECT) studies demonstrated that the small NRP-1-targeting peptides 99mTc-MA-ATWLPPR and 99mTc-CK3 showed poor tumor imaging quality, because of their rapid blood clearance and very low tumor uptake. Compared with small peptides, monoclonal antibodies (mAbs) can improve imaging of NRP-1-expression, due to their high affinity, specificity and slow extraction. A6-11-26 is a novel monoclonal antibody against NRP-1 b1b2 domain that exhibits inhibition of tumor growth in NPR-1-expressing preclinical models. The aim of the present study was to develop the 131I-labeled anti-NRP-1 monoclonal antibody A6-11-26 as a SPECT probe for imaging of NRP-1-positive tumor. An anti-NRP-1 monoclonal antibody (A6-11-26) was produced by hybridomas and was labeled with iodine-131 by the iodogen method. In vitro, the radiolabeling efficiency, radiochemical purity, immunoreactive fraction and stability were assessed. Binding affinity and specificity of 131I‑A6-11-26 to NRP-1 were evaluated using human glioblastoma U87MG cells. In vivo, biodistribution and SPECT/CT studies were conducted on mice bearing U87MG xenografts after the injection of 131I-A6-11-26 with or without co-injection of unlabeled A6-11-26 antibody. A6-11-26 was generated successfully by hybridoma with high purity (>95%) and was labeled with iodine-131 within 60 min with high labelling efficiency (95.46±3.34%), radiochemical purity (98.23±1.41%). 131I-A6-11-26 retained its immunoreactivity and also displayed excellent stability in mouse serum and PBS solution during 1 to 96 h. Cell uptake assays showed high NRP-1-specific uptake (15.80±1.30% applied activity at 6 h) in U87MG cells. 131I-A6-11-26 bound to NRP-1 with low nanomolar affinity (KD=1.67±0.14 nM) in U87MG cells. In vivo, biodistribution study demonstrated targeting of U87MG glioma xenografts was NRP-1 specific. The tumor uptake was 6.00±1.24%ID/g at 24 h, and the tumor to muscle ratio was 3.20±0.30 at 24 h, and reached the highest level of 6.13±0.24 at 120 h after injection. SPECT imaging studies revealed that 131I-A6-11-26 could clearly identify U87MG tumors with good contrast, especially at 72-120 h after injection. The present study demonstrates that 131I-A6-11-26 is capable of detecting lesions in an NRP-1-expressing tumor with high target selectivity, and may offer a promising SPECT agent for NRP-1 expression positive tumor and encourage further investigation.
Department of Nuclear Medicine, Zhongshan Hospital Xiamen University, Xiamen, Fujian 361004, P.R. China.
Xiaofeng
X
Dou
Cancer Research Center of Medical School, Xiamen University, Xiamen, Fujian 361102, P.R. China.
Jianghua
J
Yan
Department of Nuclear Medicine, Zhongshan Hospital Xiamen University, Xiamen, Fujian 361004, P.R. China.
Yafei
Y
Zhang
Department of Nuclear Medicine, Zhongshan Hospital Xiamen University, Xiamen, Fujian 361004, P.R. China.
Peng
P
Liu
Department of Nuclear Medicine, Zhongshan Hospital Xiamen University, Xiamen, Fujian 361004, P.R. China.
Yizhen
Y
Jiang
Cancer Research Center of Medical School, Xiamen University, Xiamen, Fujian 361102, P.R. China.
Sha
S
Lv
Cancer Research Center of Medical School, Xiamen University, Xiamen, Fujian 361102, P.R. China.
Fanwei
F
Zeng
Cancer Research Center of Medical School, Xiamen University, Xiamen, Fujian 361102, P.R. China.
Xiaoli
X
Chen
Cancer Research Center of Medical School, Xiamen University, Xiamen, Fujian 361102, P.R. China.
Shengyu
S
Wang
Cancer Research Center of Medical School, Xiamen University, Xiamen, Fujian 361102, P.R. China.
Haipeng
H
Zhang
Department of Nuclear Medicine, The First Affiliated Hospital of Xiamen University, Xiamen, Fujian 361003, P.R. China.
Hua
H
Wu
Department of Nuclear Medicine, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310052, P.R. China.
Hong
H
Zhang
Department of Imaging Medicine, Affiliated Southeast Hospital of Xiamen University, Zhangzhou, Fujian 363000, P.R. China.
Lin
L
Ouyang
Department of Nuclear Medicine, Zhongshan Hospital Xiamen University, Xiamen, Fujian 361004, P.R. China.
Xinhui
X
Su
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics; D000918:Antibody Specificity; D001932:Brain Neoplasms / Q000000981:diagnostic imaging* / Q000378:metabolism; D045744:Cell Line, Tumor; D003952:Diagnostic Imaging; D005909:Glioblastoma / Q000000981:diagnostic imaging* / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes / Q000737:chemistry; D051379:Mice; D009368:Neoplasm Transplantation; D039942:Neuropilin-1 / Q000276:immunology*; D019275:Radiopharmaceuticals / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics; D014018:Tissue Distribution; D015899:Tomography, Emission-Computed, Single-Photon / Q000379:methods*
D000911:Antibodies, Monoclonal; D007457:Iodine Radioisotopes; D019275:Radiopharmaceuticals; D039942:Neuropilin-1