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Quantitative radioimmunoPET imaging of EphA2 in tumor-bearing mice

  • Molecular imaging
  • Published:
European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

EphA2 receptor tyrosine kinase is significantly overexpressed in a wide variety of cancer types. High EphA2 expression has been correlated with increased metastatic potential and poor patient survival. Although many recent reports have focused on blocking the EphA2 signaling pathway in cancer, the in vivo imaging of EphA2 has not yet been investigated.

Methods

We labeled 1C1, a humanized monoclonal antibody against both human and murine EphA2, with 64Cu through the chelating agent 1,4,7,10-tetraazacyclododecane N,N′,N″,N″′-tetraacetic acid (DOTA) and carried out positron emission tomography (PET) imaging of eight tumor models with different EphA2 expression levels. Western blotting of tumor tissue lysate was performed to correlate the EphA2 expression level with 64Cu-DOTA-1C1 uptake in the tumors. Immunofluorescence staining and biodistribution studies were also carried out to validate the in vivo results.

Results

The radiolabeling yield was 88.9 ± 9.5% (n = 7) and the specific activity of 64Cu-DOTA-1C1 was 1.32 ± 0.14 GBq/mg of 1C1 mAb. The antibody retained antigen-binding affinity/specificity after DOTA conjugation as measured by FACS analysis. The uptake of 64Cu-DOTA-1C1 in CT-26 tumors was as high as 25.1 ± 2.5 %ID/g (n = 3) at 18 h post injection. 64Cu-DOTA-IgG, an isotype-matched control, exhibited minimal non-specific uptake in all eight tumor models. In vivo EphA2 specificity of 64Cu-DOTA-1C1 was confirmed by successful blocking of CT-26 tumor uptake by unlabeled 1C1. Most importantly, the tumor uptake value obtained from PET imaging had excellent linear correlation with the relative tumor tissue EphA2 expression level measured by Western blot, where r 2 equals 0.90 and 0.92 at 18 h and 42 h post injection, respectively.

Conclusion

The tumor uptake of 64Cu-DOTA-1C1 measured by microPET imaging reflects tumor EphA2 expression level in vivo. This is, to our knowledge, the first report of quantitative radioimmunoPET imaging of EphA2 in living subjects. Future clinical investigation of 64Cu-DOTA-1C1 is warranted.

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Acknowledgements

This project was financially supported by MedImmune, Inc., National Institute of Biomedical Imaging and Bioengineering (NIBIB) (R21 EB001785), National Cancer Institute (NCI) (R21 CA102123, P50 CA114747, U54 CA119367, and R24 CA93862), Department of Defense (DOD) (W81XWH-04-1-0697, W81XWH-06-1-0665, W81XWH-06-1-0042, and DAMD17-03-1-0143), and a Benedict Cassen Postdoctoral Fellowship from the Education and Research Foundation of the Society of Nuclear Medicine (to W. Cai). We thank Lina He for her excellent technical support and the cyclotron team at University of Wisconsin, Madison for 64Cu production.

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Authors and Affiliations

  1. Department of Radiology and Bio-X Program, The Molecular Imaging Program at Stanford (MIPS), Stanford University School of Medicine, 1201 Welch Rd, P095, Stanford, CA, 94305-5484, USA

    Weibo Cai, Alireza Ebrahimnejad, Kai Chen, Qizhen Cao, Zi-Bo Li & Xiaoyuan Chen

  2. MedImmune, Inc., Gaithersburg, MD, USA

    David A. Tice

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  1. Weibo Cai
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  2. Alireza Ebrahimnejad
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  7. Xiaoyuan Chen
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Correspondence to Xiaoyuan Chen.

Additional information

Weibo Cai and Alireza Ebrahimnejad contributed equally to this work.

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Cai, W., Ebrahimnejad, A., Chen, K. et al. Quantitative radioimmunoPET imaging of EphA2 in tumor-bearing mice. Eur J Nucl Med Mol Imaging 34, 2024–2036 (2007). https://doi.org/10.1007/s00259-007-0503-5

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  • DOI: https://doi.org/10.1007/s00259-007-0503-5

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