A Comparison of [99mTc]Duramycin and [99mTc]Annexin V in SPECT/CT Imaging Atherosclerotic Plaques
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Apoptosis is a key factor in unstable plaques. The aim of this study is to evaluate the utility of visualizing atherosclerotic plaques with radiolabeled duramycin and Annexin V.
ApoE−/− mice were fed with a high-fat diet to develop atherosclerosis, C57 mice as a control. Using a routine conjugation protocol, highly pure [99mTc]duramycin and [99mTc]Annexin V were obtained, which were applied for in vitro cell assays of apoptosis and in vivo imaging of atherosclerotic plaques in the animal model. Oil Red O staining, TUNEL, hematoxylin-eosin (HE), and CD68 immunostaining were used to evaluate the deposition of lipids and presence of apoptotic macrophages in the lesions where focal intensity positively correlated with the uptake of both tracers.
[99mTc]duramycin and [99mTc]Annexin V with a high radiochemical purity (97.13 ± 1.52 and 94.94 ± 0.65 %, respectively) and a well stability at room temperature were used. Apoptotic cells binding activity to [99mTc]duramycin (Kd, 6.92 nM and Bmax, 56.04 mol/1019 cells) was significantly greater than [99mTc]Annexin V (Kd, 12.63 nM and Bmax, 31.55 mol/1019 cells). Compared with [99mTc]Annexin V, [99mTc]duramycin bound avidly to atherosclerotic lesions with a higher plaque-to-background ratio (P/B was 8.23 ± 0.91 and 5.45 ± 0.48 at 20 weeks, 15.02 ± 0.23 and 12.14 ± 0.22 at 30 weeks). No plaques were found in C57 control mice. Furthermore, Oil Red O staining showed lipid deposition areas were significantly increased in ApoE−/− mice at 20 and 30 weeks, and TUNEL and CD68 staining confirmed that the focal uptake of both tracers contained abundant apoptotic macrophages.
This stable, fast clearing, and highly specific [99mTc]duramycin, therefore, can be useful for the quantification of vulnerable atherosclerotic plaques.
Key WordsAtherosclerosis Plaque vulnerability Duramycin Annexin V Phosphatidylethanolamine Phosphatidylserine Micro-SPECT/CT
Funding support from The National Nature Science Foundation of China (81471706, and 81671735) and The Shanghai Science and Technology Committee International Collaboration Project (16410722700) is gratefully appreciated. The authors are grateful to technical supports from Prof. Yingjian Zhang and Dr. Jianping Zhang from Center for Biomedical Imaging, Fudan University, and Shanghai Engineering Research Center of Molecular Imaging Probes.
Y.H., G.B.L, and D.F.C participated in the experimental and data analysis. Y.H. wrote the manuscript. Y.H., G.B.L, Y.L.L, and H.Z. involved in animal imaging and dealt with the aortas. B.G, K.P, and H.S.C. helped in polishing the articles. Y.H., D.F.C, and H.C.S designed and controlled the quality of study. All authors have reviewed the manuscript.
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Conflict of Interest
Brian D Gray and Koon Y Pak are employees of Molecular Targeting Technologies, Inc. All other authors declare that there is no conflict of interest regarding the publication of this paper. At all times, the other authors had full control over study data and interpretation.
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