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Comparative evaluation of synthetic anti-HER2 Affibody molecules site-specifically labelled with 111In using N-terminal DOTA, NOTA and NODAGA chelators in mice bearing prostate cancer xenografts

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Abstract

Purpose

In disseminated prostate cancer, expression of human epidermal growth factor receptor type 2 (HER2) is one of the pathways to androgen independence. Radionuclide molecular imaging of HER2 expression in disseminated prostate cancer might identify patients for HER2-targeted therapy. Affibody molecules are small (7 kDa) targeting proteins with high potential as tracers for radionuclide imaging. The goal of this study was to develop an optimal Affibody-based tracer for visualization of HER2 expression in prostate cancer.

Methods

A synthetic variant of the anti-HER2 ZHER2:342 Affibody molecule, ZHER2:S1, was N-terminally conjugated with the chelators DOTA, NOTA and NODAGA. The conjugated proteins were biophysically characterized by electrospray ionization mass spectroscopy (ESI-MS), circular dichroism (CD) spectroscopy and surface plasmon resonance (SPR)-based biosensor analysis. After labelling with 111In, the biodistribution was assessed in normal mice and the two most promising conjugates were further evaluated for tumour targeting in mice bearing DU-145 prostate cancer xenografts.

Results

The HER2-binding equilibrium dissociation constants were 130, 140 and 90 pM for DOTA-ZHER2:S1, NOTA-ZHER2:S1 and NODAGA-ZHER2:S1, respectively. A comparative study of 111In-labelled DOTA-ZHER2:S1, NOTA-ZHER2:S1 and NODAGA-ZHER2:S1 in normal mice demonstrated a substantial influence of the chelators on the biodistribution properties of the conjugates. 111In-NODAGA-ZHER2:S1 had the most rapid clearance from blood and healthy tissues. 111In-NOTA-ZHER2:S1 showed high hepatic uptake and was excluded from further evaluation. 111In-DOTA-ZHER2:S1 and 111In-NODAGA-ZHER2:S1 demonstrated specific uptake in DU-145 prostate cancer xenografts in nude mice. The tumour uptake of 111In-NODAGA-ZHER2:S1, 5.6 ± 0.4%ID/g, was significantly lower than the uptake of 111In-DOTA-ZHER2:S1, 7.4 ± 0.5%ID/g, presumably because of lower bioavailability due to more rapid clearance. 111In-NODAGA-ZHER2:S1 provided higher tumour-to-blood ratio, but somewhat lower tumour-to-liver, tumour-to-spleen and tumour-to-bone ratios.

Conclusion

Since distant prostate cancer metastases are situated in bone or bone marrow, the higher tumour-to-bone ratio is the most important. This renders 111In-DOTA-ZHER2:S1 a preferable agent for imaging of HER2 expression in disseminated prostate cancer.

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Acknowledgement

This study was supported by grants from the Swedish Cancer Society (Cancerfonden), the Swedish Research Council (Vetenskapsrådet) and the VINNOVA SAMBIO program.

Conflicts of interest

None.

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Authors and Affiliations

  1. Preclinical PET Platform, Department of Medicinal Chemistry, Uppsala University, Uppsala, Sweden

    Jennie Malmberg, Zohreh Varasteh & Anna Orlova

  2. Division of Molecular Biotechnology, School of Biotechnology, KTH Royal Institute of Technology, AlbaNova University Centre, 106 91, Stockholm, Sweden

    Anna Perols, Alexis Braun & Amelie Eriksson Karlström

  3. Unit of Biomedical Radiation Sciences, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden

    Mohamed Altai & Vladimir Tolmachev

  4. Section of Medical Physics, Department of Oncology, Uppsala University Hospital, Uppsala, Sweden

    Mattias Sandström

  5. Department of Medical Sciences, Section of Nuclear Medicine, Uppsala University Hospital, 751 85, Uppsala, Sweden

    Ulrike Garske

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  1. Jennie Malmberg
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Correspondence to Amelie Eriksson Karlström.

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Jennie Malmberg and Anna Perols contributed equally to this study.

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Malmberg, J., Perols, A., Varasteh, Z. et al. Comparative evaluation of synthetic anti-HER2 Affibody molecules site-specifically labelled with 111In using N-terminal DOTA, NOTA and NODAGA chelators in mice bearing prostate cancer xenografts. Eur J Nucl Med Mol Imaging 39, 481–492 (2012). https://doi.org/10.1007/s00259-011-1992-9

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  • DOI: https://doi.org/10.1007/s00259-011-1992-9

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