Development of a radiolabeled probe for detecting membrane type-1 matrix metalloproteinase on malignant tumors.
Membrane type-1 matrix metalloproteinase (MT1-MMP) expressed on the tumor cell surface activates pro-MMP-2 and pro-MMP-13 to exacerbate the malignancy, suggesting its suitability as a target molecule for diagnosis by in vivo molecular imaging. Thus, we prepared radiolabeled anti-MT1-MMP monoclonal antibody (mAb) as a novel radiolabeled probe for detecting MT1-MMP in vivo and evaluated its usefulness in breast tumor-bearing rodents. (99m)Tc-anti-MT1-MMP mAb was prepared using HYNIC as a bifunctional chelating agent and immunoreactivity was evaluated by flow cytometry. MT1-MMP expression in breast carcinoma cells (rat: Walker-256 and MRMT-1, mouse: FM3A) was measured by Western blotting. In vivo biodistribution was examined for 48 h using tumor-implanted rodents followed by estimation of radiation absorbed by a standard quantitation platform Organ Level Internal Dose Assessment (OLINDA). (99m)Tc-anti-MT1-MMP mAb was obtained with 84% immunoreactivity to MT1-MMP and more than 92% radiochemical purity. MT1-MMP was highly expressed in all malignant cells. Tumor radioactivity increased with time after administration and reached 3 to 5 times higher values at 24 h post-injection than those at 1 h. Other organs, including the stomach, showed decreasing values over time. Tumor to blood ratios increased with time and reached more than 1.3 at 48 h. The effective dose was <5.0 muSv/MBq. The results suggest that (99m)Tc-anti-MT1-MMP mAb is a promising probe for future diagnosis of breast tumors by in vivo nuclear medical imaging.
Temma
Takashi
T
Department of Patho-Functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto University.
Sano
Kohei
K
Kuge
Yuji
Y
Kamihashi
Junko
J
Takai
Nozomi
N
Ogawa
Yuki
Y
Saji
Hideo
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000737:chemistry; D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000000981:diagnostic imaging* / Q000201:enzymology* / Q000473:pathology; D045744:Cell Line, Tumor; D003947:Diagnostic Techniques, Radioisotope; D005260:Female; D006834:Hydrazines / Q000008:administration & dosage / Q000737:chemistry; D053511:Matrix Metalloproteinase 14 / Q000032:analysis*; D051379:Mice; D008809:Mice, Inbred C3H; D009368:Neoplasm Transplantation; D009539:Nicotinic Acids / Q000008:administration & dosage / Q000737:chemistry; D011877:Radionuclide Imaging; D051381:Rats; D017207:Rats, Sprague-Dawley; D013667:Technetium / Q000008:administration & dosage / Q000737:chemistry
C120392:6-hydrazinopyridine-3-carboxylic acid; D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D006834:Hydrazines; D009539:Nicotinic Acids; D013667:Technetium; D053511:Matrix Metalloproteinase 14