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ImmunoPET Imaging of αvβ6 Expression Using an Engineered Anti-αvβ6 Cys-diabody Site-Specifically Radiolabeled with Cu-64: Considerations for Optimal Imaging with Antibody Fragments

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Abstract

Purpose

Increased expression of the αvβ6 integrin correlates with advanced tumor grade and poor clinical outcome, identifying αvβ6 as a prognostic indicator and an attractive target for molecular imaging. This work investigated the ability of a disulfide-stabilized [64Cu]NOTA-αvβ6 cys-diabody to image αvβ6 expression in vivo using a nu/nu mouse model bearing human melanoma xenografts and positron-emission tomography.

Procedures

Small-animal positron emission tomography (PET) imaging, quantitative ROI analysis, and ex vivo biodistribution were conducted to ascertain tumor uptake and organ distribution of the [64Cu]NOTA-αvβ6 cys-diabody. Immunohistochemical staining of tumors and mouse organs and immunoreactivity assays were utilized to correlate in vivo and ex vivo observations.

Results

PET imaging of the [64Cu]NOTA-αvβ6 cys-diabody revealed low tumor uptake at 24 h p.i. in DX3Puroβ6 tumors (2.69 ± 0.45 %ID/g) with comparable results found in the DX3Puro tumors (2.24 ± 0.15 %ID/g). Quantitative biodistribution confirmed that DX3Puroβ6 tumor uptake was highest at 24 h p.i. (4.63 ± 0.18 %ID/g); however, uptake was also observed in the stomach (4.84 ± 2.99 %ID/g), small intestines (4.50 ± 1.69 %ID/g), large intestines (4.73 ± 0.97 %ID/g), gallbladder (6.04 ± 1.88 %ID/g), and lungs (3.89 ± 0.69 %ID/g).

Conclusions

Small-animal PET imaging was successful in visualizing αvβ6-positive tumor uptake of the [64Cu]NOTA-αvβ6 cys-diabody. Cys-diabody cross-reactivity was observed between human and murine αvβ6 and immunohistochemical staining confirmed the presence of an endogenous αvβ6 antigen sink, which led to suboptimal tumor contrast in this mouse model. Future investigations will focus on dose escalation studies to overcome the endogenous antigen sink while increasing DX3Puroβ6 tumor uptake.

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Acknowledgments

The authors thank the staff at CMGI, including Dave Kukis, Jennifer Fung, and Charles Smith, for their assistance with radioisotope procurement and animal handling during the PET imaging studies. Also, a special thanks to Dr. Anna Wu and Dr. Kirstin Zettlitz at the Crump Institute for Molecular Imaging, UCLA, for many fruitful discussions and analyses.

Author information

Authors and Affiliations

  1. Department of Biomedical Engineering, University of California, Davis, Davis, CA, USA

    Jason B. White, Lina Y. Hu, David L. Boucher & Julie L. Sutcliffe

  2. Division of Hematology and Oncology, Department of Internal Medicine, University of California, Davis, Sacramento, CA, USA

    Julie L. Sutcliffe

  3. Center for Molecular and Genomic Imaging, University of California, Davis, Davis, CA, USA

    Julie L. Sutcliffe

  4. Radiochemistry Research and Training Facility, University of California, Davis, Sacramento, CA, USA

    Julie L. Sutcliffe

Authors
  1. Jason B. White
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  2. Lina Y. Hu
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  4. Julie L. Sutcliffe
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Corresponding author

Correspondence to Julie L. Sutcliffe.

Ethics declarations

All animal handling and procedures were performed using protocols approved by the UC Davis Institutional Animal Care and Use Committee.

Conflict of Interest

The authors declare that they have no conflict of interest.

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White, J.B., Hu, L.Y., Boucher, D.L. et al. ImmunoPET Imaging of αvβ6 Expression Using an Engineered Anti-αvβ6 Cys-diabody Site-Specifically Radiolabeled with Cu-64: Considerations for Optimal Imaging with Antibody Fragments. Mol Imaging Biol 20, 103–113 (2018). https://doi.org/10.1007/s11307-017-1097-3

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