Skip to main content

Advertisement

SpringerLink
Log in

Imaging of HER3-expressing xenografts in mice using a 99mTc(CO)3-HEHEHE-ZHER3:08699 affibody molecule

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

    We’re sorry, something doesn't seem to be working properly.

    Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Abstract

Purpose

Human epidermal growth factor receptor type 3 (HER3) is a transmembrane receptor tyrosine kinase belonging to the HER (ErbB) receptor family. Membranous expression of HER3 is associated with trastuzumab resistance in breast cancer and the transition to androgen independence in prostate cancer. Imaging of HER3 expression in malignant tumors may provide important diagnostic information that can influence patient management. Affibody molecules with low picomolar affinity to HER3 were recently selected. The aim of this study was to investigate the feasibility of HER3 imaging using radiolabeled Affibody molecules.

Methods

A HER3-binding Affibody molecule, Z08699, with a HEHEHE-tag on N-terminus was labeled with 99mTc(CO)3 using an IsoLink kit. In vitro and in vivo binding specificity and the cellular processing of the labeled binder were evaluated. Biodistribution of 99mTc(CO)3-HEHEHE-Z08699 was studied over time in mice bearing HER3-expressing xenografts.

Results

HEHEHE-Z08699 was labeled with 99mTc(CO)3 with an isolated yield of >80 % and a purity of >99 %. Binding of 99mTc(CO)3-HEHEHE-Z08699 was specific to BT474 and MCF7 (breast cancer), and LS174T (colon cancer) cells. Cellular processing showed rapid binding and relatively quick internalization of the receptor/Affibody molecule complex (70 % of cell-associated radioactivity was internalized after 24 h). The tumor targeting was receptor mediated and the excretion was predominantly renal. Receptor-mediated uptake was also found in the liver, lung, stomach, intestine, and salivary glands. At 4 h pi, tumor-to-blood ratios were 7 ± 3 for BT474, and 6 ± 2 for LS174T xenografts. LS174T tumors were visualized by microSPECT 4 h pi.

Conclusions

The results of this study suggest the feasibility of HER3-imaging in malignant tumors using Affibody molecules.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Yarden Y, Sliwkowski MX. Untangling the ErbB signalling network. Nat Rev Mol Cell Biol. 2001;2:127–37.

    Article  CAS  PubMed  Google Scholar 

  2. Marmor MD, Skaria KB, Yarden Y. Signal transduction and oncogenesis by ErbB/HER receptors. Int J Radiat Oncol Biol Phys. 2004;58:903–13.

    Article  CAS  PubMed  Google Scholar 

  3. Ocaña A, Pandiella A. Targeting HER receptors in cancer. Curr Pharm Des. 2013;19:808–17.

    Article  PubMed  Google Scholar 

  4. Aurisicchio L, Marra E, Roscilli G, Mancini R, Ciliberto G. The promise of anti-ErbB3 monoclonals as new cancer therapeutics. Oncotarget. 2012;3:744–58.

    PubMed Central  PubMed  Google Scholar 

  5. Hsieh AC, Moasser MM. Targeting HER proteins in cancer therapy and the role of the non-target HER3. Br J Cancer. 2007;97:453–7.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Baselga J, Swain SM. Novel anticancer targets: revisiting ERBB2 and discovering ERBB3. Nat Rev Cancer. 2009;9:463–75.

    Article  CAS  PubMed  Google Scholar 

  7. Garrett JT, Olivares MG, Rinehart C, Granja-Ingram ND, Sánchez V, Chakrabarty A, et al. Transcriptional and posttranslational up-regulation of HER3 (ErbB3) compensates for inhibition of the HER2 tyrosine kinase. Proc Natl Acad Sci U S A. 2011;108:5021–6.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Kruser TJ, Wheeler DL. Mechanisms of resistance to HER family targeting antibodies. Exp Cell Res. 2010;316:1083–100.

    Article  CAS  PubMed  Google Scholar 

  9. Servidei T, Riccardi A, Mozzetti S, Ferlini C, Riccardi R. Chemoresistant tumor cell lines display altered epidermal growth factor receptor and HER3 signaling and enhanced sensitivity to gefitinib. Int J Cancer. 2008;123:2939–49.

    Article  CAS  PubMed  Google Scholar 

  10. Jathal MK, Chen L, Mudryj M, Ghosh PM. Targeting ErbB3: the New RTK(id) on the Prostate Cancer Block. Immunol Endocr Metab Agents Med Chem. 2011;11:131–49.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. Ocana A, Vera-Badillo F, Seruga B, Templeton A, Pandiella A, Amir E. HER3 overexpression and survival in solid tumors: a meta-analysis. J Natl Cancer Inst. 2013;105:266–73.

    Article  CAS  PubMed  Google Scholar 

  12. Tolmachev V, Stone-Elander S, Orlova A. Radiolabelled receptor-tyrosine-kinase targeting drugs for patient stratification and monitoring of therapy response: prospects and pitfalls. Lancet Oncol. 2010;11:992–1000.

    Article  CAS  PubMed  Google Scholar 

  13. van Dongen GA, Poot AJ, Vugts DJ. PET imaging with radiolabeled antibodies and tyrosine kinase inhibitors: immuno-PET and TKI-PET. Tumour Biol. 2012;33:607–15.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. van Dongen GA, Visser GW, Lub-de Hooge MN, de Vries EG, Perk LR. Immuno-PET: a navigator in monoclonal antibody development and applications. Oncologist. 2007;12:1379–89.

    Article  PubMed  Google Scholar 

  15. Wester HJ, Kessler H. Molecular targeting with peptides or peptide-polymer conjugates: just a question of size? J Nucl Med. 2005;46:1940–5.

    CAS  PubMed  Google Scholar 

  16. Löfblom J, Feldwisch J, Tolmachev V, Carlsson J, Ståhl S, Frejd FY. Affibody molecules: engineered proteins for therapeutic, diagnostic and biotechnological applications. FEBS Lett. 2010;584:2670–80.

    Article  PubMed  Google Scholar 

  17. Orlova A, Magnusson M, Eriksson TL, Nilsson M, Larsson B, Höidén-Guthenberg I, et al. Tumor imaging using a picomolar affinity HER2 binding affibody molecule. Cancer Res. 2006;66:4339–48.

    Article  CAS  PubMed  Google Scholar 

  18. Tolmachev V, Rosik D, Wållberg H, Sjöberg A, Sandström M, Hansson M, et al. Imaging of EGFR expression in murine xenografts using site-specifically labelled anti-EGFR 111In-DOTA-Z EGFR:2377 Affibody molecule: aspect of the injected tracer amount. Eur J Nucl Med Mol Imaging. 2010;37:613–22.

    Article  PubMed  Google Scholar 

  19. Tolmachev V, Malmberg J, Hofström C, Abrahmsén L, Bergman T, Sjöberg A, et al. Imaging of IGF-1R in prostate cancer xenografts using the Affibody molecule 111In-DOTA-Z IGF1R:4551. J Nucl Med. 2012;53:146–53.

    Article  PubMed  Google Scholar 

  20. Ahlgren S, Orlova A, Rosik D, Sandström M, Sjöberg A, Baastrup B, et al. Evaluation of Maleimide Derivative of DOTA for Site-Specific Labeling of Recombinant Affibody Molecules. Bioconjug Chem. 2008;19:235–43.

    Article  CAS  PubMed  Google Scholar 

  21. Friedman M, Orlova A, Johansson E, Eriksson T, Höidén-Guthenberg I, Tolmachev V, et al. Directed evolution to low nanomolar affinity of an epidermal growth factor receptor 1-binding Affibody molecule. J Mol Biol. 2008;376:1388–402.

    Article  CAS  PubMed  Google Scholar 

  22. Tolmachev V, Tran TA, Rosik D, Abrahmsén L, Sjöberg A, Orlova A. Tumor targeting using Affibody molecules: an interplay of a target expression level, affinity and binding site composition. J Nucl Med. 2012;53:953–60.

    Article  CAS  PubMed  Google Scholar 

  23. Baum RP, Prasad V, Müller D, Schuchardt C, Orlova A, Wennborg A, et al. Molecular imaging of HER2-expressing malignant tumors in breast cancer patients using synthetic 111In- or 68Ga-labeled affibody molecules. J Nucl Med. 2010;51:892–7.

    Article  PubMed  Google Scholar 

  24. Sörensen J, Sandberg D, Carlsson J, Sandström M, Wennborg A, Feldwisch J, Tolmachev V, Åström G, Lubberink M, Garske-Roman U, Lindman H. First-in-Human Molecular Imaging of HER2 Expression in Breast Cancer Metastases Using the 111In-ABY-025 Affibody Molecule. J Nucl Med, 2014, in press.

  25. Kronqvist N, Malm M, Göstring L, Gunneriusson E, Nilsson M, Höidén Guthenberg I, et al. Combining phage and staphylococcal surface display for generation of ErbB3-specific Affibody molecules. Protein Eng Des Sel. 2011;24:385–96.

    Article  CAS  PubMed  Google Scholar 

  26. Robinson MK, Hodge KM, Horak E, Sundberg AL, Russeva M, Shaller CC, et al. Targeting ErbB2 and ErbB3 with a bispecific single-chain Fv enhances targeting selectivity and induces a therapeutic effect in vitro. Br J Cancer. 2008;99:1415–25.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Löfblom J. Bacterial display in combinatorial protein engineering. Biotechnol J. 2011;6:1115–29.

    Article  PubMed  Google Scholar 

  28. Malm M, Kronqvist N, Lindberg H, Gudmundsdotter L, Bass T, Frejd FY, et al. Inhibiting HER3-mediated tumor cell growth with Affibody molecules engineered to low picomolar affinity by position-directed error-prone PCR-like diversification. PLoS One. 2013;8:e62791.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  29. Tolmachev V, Hofström C, Malmberg J, Ahlgren S, Hosseinimehr SJ, Sandström M, et al. HEHEHE-tagged affibody molecule may be purified by IMAC, is conveniently labeled with [99(m)Tc(CO)3](+), and shows improved biodistribution with reduced hepatic radioactivity accumulation. Bioconjug Chem. 2010;21:2013–22.

    Article  CAS  PubMed  Google Scholar 

  30. Hofström C, Altai M, Honarvar H, Strand J, Malmberg J, Hosseinimehr SJ, et al. HAHAHA, HEHEHE, HIHIHI or HKHKHK: influence of position and composition of histidine containing tags on biodistribution of [99mTc(CO)3] + - labeled affibody molecules. J Med Chem. 2013;56:4966–74.

    Article  PubMed  Google Scholar 

  31. Waibel R, Alberto R, Willuda J, et al. Stable one-step technetium-99 m labeling of His-tagged recombinant proteins with a novel Tc(I)-carbonyl complex. Nat Biotechnol. 1999;17:897–901.

    Article  CAS  PubMed  Google Scholar 

  32. Yoshioka T, Nishikawa Y, Ito R, Kawamata M, Doi Y, Yamamoto Y, et al. Significance of integrin αvβ5 and erbB3 in enhanced cell migration and liver metastasis of colon carcinomas stimulated by hepatocyte-derived heregulin. Cancer Sci. 2010;101:2011–8.

    Article  CAS  PubMed  Google Scholar 

  33. Janmaat ML, Rodriguez JA, Jimeno J, Kruyt FA, Giaccone G. Kahalalide F induces necrosis-like cell death that involves depletion of ErbB3 and inhibition of Akt signaling. Mol Pharmacol. 2005;68:502–10.

    CAS  PubMed  Google Scholar 

  34. Kimura F, Iwaya K, Kawaguchi T, Kaise H, Yamada K, Mukai K, et al. Epidermal growth factor-dependent enhancement of invasiveness of squamous cell carcinoma of the breast. Cancer Sci. 2010;101:1133–40.

    Article  CAS  PubMed  Google Scholar 

  35. Wållberg H, Orlova A. Slow internalization of anti-HER2 synthetic affibody monomer 111In-DOTA-ZHER2:342-pep2: implications for development of labeled tracers. Cancer Biother Radiopharm. 2008;23:435–42.

    Article  PubMed  Google Scholar 

  36. Tolmachev V, Wållberg H, Sandström M, Hansson M, Wennborg A, Orlova A. Optimal specific radioactivity of anti-HER2 Affibody molecules enables discrimination between xenografts with high and low HER2 expression levels. Eur J Nucl Med Mol Imaging. 2011;38:531–9.

    Article  PubMed  Google Scholar 

  37. Orlova A, Nilsson F, Wikman M, Widström C, Ståhl S, Carlsson J, et al. Comparative in vivo evaluation of technetium and iodine labels on an anti-HER2 Affibody for single-photon imaging of HER2 expression in tumors. J Nucl Med. 2006;47:512–9.

    CAS  PubMed  Google Scholar 

  38. Orlova A, Tolmachev V, Pehrson R, Lindborg M, Tran T, Sandström M, et al. Synthetic Affibody Molecules: A Novel Class of Affinity Ligands for Molecular Imaging of HER2-Expressing Malignant Tumors. Cancer Res. 2007;67:2178–86.

    Article  CAS  PubMed  Google Scholar 

  39. Tolmachev V, Varasteh Z, Honarvar H, Eriksson O, Jonasson P, Frejd FY, Abrahmsen L, Orlova A. Imaging of PDGFRβ expression in gliobalstoma xenografts using Affibody molecule 111In-DOTA-Z09591. J Nucl Med, 2014;55:294–300.

    Google Scholar 

  40. Orlova A, Hofström C, Strand J, Varasteh Z, Sandstrom M, Andersson K, et al. [99mTc(CO)3]+-(HE)3-ZIGF1R:4551, a new affibody conjugate for visualization of insulin-like growth factor 1 receptor expression in malignant tumours. Eur J Nucl Med Mol Imaging. 2013;40(3):439–49.

    Article  CAS  PubMed  Google Scholar 

  41. Tolmachev V. Imaging of HER-2 overexpression in tumors for guiding therapy. Curr Pharm Des. 2008;14:2999–3019.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Swedish Cancer Society (Cancerfonden) and the Swedish Research Council (Vetenskapsrådet).

Conflict of interest

The authors declare that they have no conflicts of interest.

Author information

Authors and Affiliations

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

    Anna Orlova, Maria Rosestedt, Zohreh Varasteh & Ram Kumar Selvaraju

  2. Division of Protein Technology, School of Biotechnology, KTH Royal Institute of Technology, Stockholm, Sweden

    Magdalena Malm, Ken Andersson, Stefan Ståhl & John Löfblom

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

    Mohamed Altai, Hadis Honarvar, Joanna Strand & Vladimir Tolmachev

Authors
  1. Anna Orlova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Magdalena Malm
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maria Rosestedt
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zohreh Varasteh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ken Andersson
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ram Kumar Selvaraju
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mohamed Altai
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hadis Honarvar
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Joanna Strand
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Stefan Ståhl
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Vladimir Tolmachev
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. John Löfblom
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anna Orlova.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOCX 109 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Orlova, A., Malm, M., Rosestedt, M. et al. Imaging of HER3-expressing xenografts in mice using a 99mTc(CO)3-HEHEHE-ZHER3:08699 affibody molecule. Eur J Nucl Med Mol Imaging 41, 1450–1459 (2014). https://doi.org/10.1007/s00259-014-2733-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00259-014-2733-7

Keywords

Search

Navigation