Abstract
Purpose
Targeted delivery of radionuclides for diagnostic and therapeutic applications has until recently largely been limited to receptor ligands, antibodies and antibody-derived molecules. Here, we present a new type of molecule, a 15-kDa bivalent affibody called (ZHER2:4)2, with potential for such applications. The (ZHER2:4)2 affibody showed high apparent affinity (K D=3 nM) towards the oncogene product HER-2 (also called p185/neu or c-erbB-2), which is often overexpressed in breast and ovarian cancers. The purpose of this study was to investigate the in vivo properties of the new targeting agent.
Methods
The biodistribution and tumour uptake of the radioiodinated (ZHER2:4)2 affibody was studied in nude mice carrying tumours from xenografted HER-2 overexpressing SKOV-3 cells.
Results
The radioiodinated (ZHER2:4)2 affibody was primarily excreted through the kidneys, and significant amounts of radioactivity were specifically targeted to the tumours. The blood-borne radioactivity was, at all times, mainly in the macromolecular fraction. A tumour-to-blood ratio of about 10:1 was obtained 8 h post injection, and the tumours could be easily visualised with a gamma camera at this time point.
Conclusion
The results indicate that the (ZHER2:4)2 affibody is an interesting candidate for applications in nuclear medicine, such as radionuclide-based tumour imaging and therapy.
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References
Scholl S, Beuzeboc P, Pouillart P. Targeting HER2 in other tumor types. Ann Oncol 2001;12 Suppl 1:S81–7
Berchuck A, Kamel A, Whitaker R, Kerns B, Olt G, Kinney R, et al. Overexpression of HER-2/neu is associated with poor survival in advanced epithelial ovarian cancer. Cancer Res 1990;50:4087–91
Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 1987;235:177–82
Carlomagno C, Perrone F, Gallo C, De Laurentiis M, Lauria R, Morabito A, et al. c-erb B2 overexpression decreases the benefit of adjuvant tamoxifen in early-stage breast cancer without axillary lymph node metastases. J Clin Oncol 1996;14:2702–8
Carlsson J, Nordgren H, Sjostrom J, Wester K, Villman K, Bengtsson NO, et al. HER2 expression in breast cancer primary tumours and corresponding metastases. Original data and literature review. Br J Cancer 2004;90:2344–8.
Press MF, Cordon-Cardo C, Slamon DJ. Expression of the HER-2/neu proto-oncogene in normal human adult and fetal tissues. Oncogene 1990;5:953–62
Natali PG, Nicotra MR, Bigotti A, Venturo I, Slamon DJ, Fendly BM, et al. Expression of the p185 encoded by HER2 oncogene in normal and transformed human tissues. Int J Cancer 1990;45:457–61
Klapper LN, Glathe S, Vaisman N, Hynes NE, Andrews GC, Sela M, et al. The ErbB-2/HER2 oncoprotein of human carcinomas may function solely as a shared coreceptor for multiple stroma-derived growth factors. Proc Natl Acad Sci U S A 1999;96:4995–5000
Yarden Y, Sliwkowski MX. Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2001;2:127–37.
Beaumier PL, Krohn KA, Carrasquillo JA, Eary J, Hellstrom I, Hellstrom KE, et al. Melanoma localization in nude mice with monoclonal Fab against p97. J Nucl Med 1985;26:1172–9
Hu S, Shively L, Raubitschek A, Sherman M, Williams LE, Wong JY, et al. Minibody: a novel engineered anti-carcinoembryonic antigen antibody fragment (single-chain Fv-CH3) which exhibits rapid, high-level targeting of xenografts. Cancer Res 1996;56:3055–61
Schier R, Marks JD, Wolf EJ, Apell G, Wong C, McCartney JE, et al. In vitro and in vivo characterization of a human anti-c-erbB-2 single-chain Fv isolated from a filamentous phage antibody library. Immunotechnology 1995;1:73–81
Nord K, Gunneriusson E, Ringdahl J, Stahl S, Uhlen M, Nygren PA. Binding proteins selected from combinatorial libraries of an alpha-helical bacterial receptor domain. Nat Biotechnol 1997;15:772–7
Wikman M, Steffen AC, Gunneriusson E, Tolmachev V, Adams GP, Carlsson J, et al. Selection and characterization of HER2/neu-binding affibody ligands. Protein Eng Des Sel 2004;17:455–62
Gunneriusson E, Nord K, Uhlen M, Nygren P. Affinity maturation of a Taq DNA polymerase specific affibody by helix shuffling. Protein Eng 1999;12:873–8
Steffen AC, Wikman M, Tolmachev V, Adams GP, Nilsson FY, Ståhl S, et al. In vitro characterization of a bivalent anti-HER-2 affibody with potential for radionuclide-based diagnostics.Cancer Biother Radiopharm 2005; 20:239–48
Wester K, Asplund A, Backvall H, Micke P, Derveniece A, Hartmane I, et al. Zinc-based fixative improves preservation of genomic DNA and proteins in histoprocessing of human tissues. Lab Invest 2003;83:889–99
Adams GP, Schier R, McCall AM, Crawford RS, Wolf EJ, Weiner LM, et al. Prolonged in vivo tumour retention of a human diabody targeting the extracellular domain of human HER2/neu. Br J Cancer 1998;77:1405–12
Adams GP, Schier R, McCall AM, Simmons HH, Horak EM, Alpaugh RK, et al. High affinity restricts the localization and tumor penetration of single-chain fv antibody molecules. Cancer Res 2001;61:4750–5
Adams GP. Improving the tumor specificity and retention of antibody-based molecules. In Vivo 1998;12:11–21
Mogensen CE, Solling. Studies on renal tubular protein reabsorption: partial and near complete inhibition by certain amino acids. Scand J Clin Lab Invest 1977;37:477–86
Bernard A, Ouled Amor A, Lauwerys R. Charge-dependent renal uptake of beta 2-microglobulin in conscious rats. Scand J Clin Lab Invest 1992;52:415–23
Huston JS, George AJ, Adams GP, Stafford WF, Jamar F, Tai MS, et al. Single-chain Fv radioimmunotargeting. Q J Nucl Med 1996;40:320–33
Carlsson J, Forssell Aronsson E, Hietala SO, Stigbrand T, Tennvall J. Tumour therapy with radionuclides: assessment of progress and problems. Radiother Oncol 2003;66:107–17
Weinstein JN, Eger RR, Covell DG, Black CD, Mulshine J, Carrasquillo JA, et al. The pharmacology of monoclonal antibodies. Ann N Y Acad Sci 1987;507:199–210
Fujimori K, Covell DG, Fletcher JE, Weinstein JN. Modeling analysis of the global and microscopic distribution of immunoglobulin G, F(ab')2, and Fab in tumors. Cancer Res 1989;49:5656–63
Lindstrom A, Carlsson J. Penetration and binding of epidermal growth factor-dextran conjugates in spheroids of human glioma origin. Cancer Biother 1993;8:145–58
Tolmachev V, Orlova A, Lundqvist H. Approaches to improve cellular retention of radiohalogen labels delivered by internalising tumour-targeting proteins and peptides. Curr Med Chem 2003;10:2447–60
Sundberg AL, Blomquist E, Carlsson J, Steffen AC, Gedda L. Cellular retention of radioactivity and increased radiation dose. Model experiments with EGF-dextran. Nucl Med Biol 2003;30:303–15
Mattes MJ, Griffiths GL, Diril H, Goldenberg DM, Ong GL, Shih LB. Processing of antibody-radioisotope conjugates after binding to the surface of tumor cells. Cancer 1994;73:787–93.
Motta-Hennessy C, Sharkey RM, Goldenberg DM. Metabolism of indium-111-labeled murine monoclonal antibody in tumor and normal tissue of the athymic mouse. J Nucl Med 1990;31:1510–9
Shih LB, Thorpe SR, Griffiths GL, Diril H, Ong GL, Hansen HJ, et al. The processing and fate of antibodies and their radiolabels bound to the surface of tumor cells in vitro: a comparison of nine radiolabels. J Nucl Med 1994;35:899–908
Acknowledgements
The authors thank Charles Widström at the department of Nuclear Medicine at Uppsala University Hospital for help with the gamma camera. Thanks also to Kenneth Wester and Veronika Asplund Eriksson for help with the immunohistochemistry, autoradiography and photographic procedures. The work was financially supported by grant no 0980-B04-17XCC (040171) from the Swedish Cancer Society.
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Steffen, AC., Orlova, A., Wikman, M. et al. Affibody-mediated tumour targeting of HER-2 expressing xenografts in mice. Eur J Nucl Med Mol Imaging 33, 631–638 (2006). https://doi.org/10.1007/s00259-005-0012-3
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DOI: https://doi.org/10.1007/s00259-005-0012-3