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Development of a radiolabeled probe for detecting membrane type-1 matrix metalloproteinase on malignant tumors
Takashi Temma1, Kohei Sano, Yuji Kuge
1Department of Patho-Functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto University.
Abstract:
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.
Insights
A new technetium-99m labeled anti-membrane type-1 matrix metalloproteinase (MT1-MMP) antibody shows promise for breast tumor imaging. This radiolabeled probe effectively detects MT1-MMP in vivo, offering a potential new tool for nuclear medical diagnostics.
Area of Science:
- Oncology
- Radiochemistry
- Molecular Imaging
Background:
- Membrane type-1 matrix metalloproteinase (MT1-MMP) promotes tumor malignancy by activating pro-MMPs.
- MT1-MMP's surface expression makes it a potential target for in vivo molecular imaging.
Purpose of the Study:
- To develop and evaluate a novel radiolabeled probe, (99m)Tc-anti-MT1-MMP mAb, for detecting MT1-MMP in vivo.
- To assess the probe's utility in breast tumor-bearing rodents for diagnostic purposes.
Main Methods:
- Preparation of (99m)Tc-anti-MT1-MMP mAb using HYNIC bifunctional chelating agent.
- Evaluation of immunoreactivity via flow cytometry and MT1-MMP expression in cancer cells using Western blotting.
- In vivo biodistribution studies in tumor-implanted rodents over 48 hours, with radiation dose estimation using OLINDA.
Main Results:
- The probe achieved 84% immunoreactivity and >92% radiochemical purity.
- High MT1-MMP expression was confirmed in all tested malignant cells.
- Tumor radioactivity increased significantly over time, with tumor-to-blood ratios exceeding 1.3 at 48 hours.
- Low effective radiation dose (<5.0 μSv/MBq) was observed.
Conclusions:
- (99m)Tc-anti-MT1-MMP mAb demonstrates high specificity and favorable biodistribution for MT1-MMP detection.
- The developed radiolabeled antibody is a promising candidate for future in vivo nuclear medical imaging of breast tumors.
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