In vivo imaging of radiation-induced tissue apoptosis by Tc(I)-his-annexin A5.

Objective: A recombinant annexin A5 with the N-terminal extension of six histidine residues was labeled with Tc(I)-tricarbonyl ion to produce the Tc-labeled annexin A5, referred to Tc(I)-his-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: [Tc(CO)(OH)] was prepared and taken to directly label his-annexin A5. The radiochemical purity of Tc(I)-his-annexin A5 after size-exclusion separation was measured by HPLC. The binding affinity of Tc(I)-his-annexin A5 to apoptotic cells was assessed using 20 Gy-irradiated Jurkat T cells. The effectiveness of Tc(I)-his-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 Tc(I)-his-annexin A5 could attain ≥95%. The binding affinity of Tc(I)-his-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 Tc(I)-his-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 Tc(I)-his-annexin A5 is a potential clinical imaging agent for detecting radiation-induced tissue apoptosis in an animal model. [ABSTRACT FROM AUTHOR]