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In vivo imaging of radiation-induced tissue apoptosis by 99mTc(I)-his6-annexin A5

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Abstract

Objective

A recombinant annexin A5 with the N-terminal extension of six histidine residues was labeled with 99mTc(I)-tricarbonyl ion to produce the 99mTc-labeled annexin A5, referred to 99mTc(I)-his6-annexin A5. We have explored the agent as an effective imaging probe for in vivo detecting the apoptosis of internal tissue subjected with high radiation doses in a γ-irradiated mouse model.

Methods

[99mTc(CO)3(OH2)3]+ was prepared and taken to directly label his6-annexin A5. The radiochemical purity of 99mTc(I)-his6-annexin A5 after size-exclusion separation was measured by HPLC. The binding affinity of 99mTc(I)-his6-annexin A5 to apoptotic cells was assessed using 20 Gy-irradiated Jurkat T cells. The effectiveness of 99mTc(I)-his6-annexin A5 as an imaging probe to detect the internal tissue apoptosis was assessed by biodistribution study and nanoSPECT/CT using the animal model of C57BL/6J mice conducted with 10 Gy γ irradiation.

Results

The radiochemical purity of 99mTc(I)-his6-annexin A5 could attain ≥95%. The binding affinity of 99mTc(I)-his6-annexin A5 to the 20 Gy-irradiated Jurkat cells was found to be ca. 20-fold higher than that to the sham-irradiated cells. In the animal imaging study, the splenic uptake of 99mTc(I)-his6-annexin A5 for the 10 Gy-irradiated mice showed from ca. 3-fold to 5-fold higher than those of the sham-irradiated mice from 45 to 165 min postinjection. The corresponding intestinal uptake showed from ca. 2-fold to 3-fold higher during the same period of time postinjection. The biodistribution study demonstrated the organ uptakes comparable with the imaging results. The apoptotic extents of the spleen and the intestine from the SPECT/CT imaging were correlated with an immunohistochemical staining assay for caspase 3 active form fragment.

Conclusion

This work is the first study to demonstrate that 99mTc(I)-his6-annexin A5 is a potential clinical imaging agent for detecting radiation-induced tissue apoptosis in an animal model.

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Acknowledgments

This study was financially supported by Chang-Gung/Tsing-Hua Collaborative Research Project. We would like to express our sincere appreciation to Professor Shiaw-Pyng Wey of Department of Medical Imaging and Radiological Sciences, Chang Gung University, Linkou, Taogyuan, Taiwan for his help in preparation of 99mTc-labeled annexin A5. The advice to the paper from Professor Chien-Wen Chang of Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, Taiwan is very much appreciated. We thank Ms. Yi-Ching Lin for the technical assistance to the study.

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Authors and Affiliations

  1. Molecular Imaging Center and Department of Nuclear Medicine, Chang Gung Memorial Hospital, Taoyuan, 33305, Taiwan

    Kun-Ju Lin

  2. Department of Medical Imaging and Radiological Sciences, Chang Gung University, Taoyuan, 33305, Taiwan

    Kun-Ju Lin

  3. Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, 30013, Taiwan

    Chun-Chung Wu, Yi-Hsin Pan, Fang-Hsing Chen, Sheng-Yung Fu, Chi-Shiun Chiang, Ji-Hong Hong & Jem-Mau Lo

  4. Department of Radiation Oncology, Chang Gung Memorial Hospital, Linkou, Taoyuan, 33305, Taiwan

    Fang-Hsing Chen & Sheng-Yung Fu

  5. Department of Nuclear Engineering and Science, National Tsing Hua University, Hsinchu, 30013, Taiwan

    Jem-Mau Lo

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  1. Kun-Ju Lin
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Correspondence to Jem-Mau Lo.

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Lin, KJ., Wu, CC., Pan, YH. et al. In vivo imaging of radiation-induced tissue apoptosis by 99mTc(I)-his6-annexin A5. Ann Nucl Med 26, 272–280 (2012). https://doi.org/10.1007/s12149-012-0571-x

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  • DOI: https://doi.org/10.1007/s12149-012-0571-x

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