nach oben

European Journal of Nuclear Medicine and Molecular Imaging

Erschienen in:

01.11.2007 | Original Article

^99mTc-chelator engineering to improve tumour targeting properties of a HER2-specific Affibody molecule

verfasst von: Torun Engfeldt, Thuy Tran, Anna Orlova, Charles Widström, Joachim Feldwisch, Lars Abrahmsen, Anders Wennborg, Amelie Eriksson Karlström, Vladimir Tolmachev

Erschienen in: European Journal of Nuclear Medicine and Molecular Imaging | Ausgabe 11/2007

Einloggen, um Zugang zu erhalten

Abstract

Purpose

Monitoring HER2 expression is crucial for selection of breast cancer patients amenable to HER2-targeting therapy. The Affibody molecule Z_HER2:342 binds to HER2 with picomolar affinity and enables specific imaging of HER2 expression. Previously, Z_HER2:342 with the additional N-terminal mercaptoacetyl-glycyl-glycyl-glycyl (maGGG) sequence was labelled with ^99mTc and demonstrated specific targeting of HER2-expressing xenografts. However, hepatobiliary excretion caused high radioactivity accumulation in the abdomen. We investigated whether the biodistribution of Z_HER2:342 can be improved by substituting glycyl residues in the chelating sequence with more hydrophilic seryl residues.

Methods

The Affibody molecule Z_HER2:342, carrying the chelators mercaptoacetyl-glycyl-seryl-glycyl (maGSG), mercaptoacetyl-glycyl-D-seryl-glycyl [maG(D-S)G] and mercaptoacetyl-seryl-seryl-seryl (maSSS), were prepared by peptide synthesis and labelled with ^99mTc. The differences in the excretion pathways were evaluated in normal mice. The tumour targeting capacity of ^99mTc-maSSS-Z_HER2:342 was studied in nude mice bearing SKOV-3 xenografts and compared with the capacity of radioiodinated Z_HER2:342.

Results

A shift towards renal excretion was obtained when glycine was substituted with serine in the chelating sequence. The radioactivity in the gastrointestinal tract was reduced threefold for the maSSS conjugate in comparison with the maGGG conjugate 4 h post injection (p.i.). The tumour uptake of ^99mTc-maSSS-Z_HER2:342 was 11.5 ± 0.5% IA/g 4 h p.i., and the tumour-to-blood ratio was 76. The pharmacokinetics and uptake characteristics of technetium-labelled Z_HER2:342 were better than those of radioiodinated Z_HER2:342.

Conclusion

The introduction of serine residues in the chelator results in better tumour imaging properties of the Affibody molecule Z_HER2:342 compared with glycyl-containing chelators and is favourable for imaging of tumours and metastases in the abdominal area.

Aunoble B, Sanches R, Didier E, Bignon YJ. Major oncogenes and tumor suppressor genes involved in epithelial ovarian cancer (review). Int J Oncol 2000;16:567–76.PubMed

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.PubMed

Gardmark T, Wester K, De la Torre M, Carlsson J, Malmstrom PU. Analysis of HER2 expression in primary urinary bladder carcinoma and corresponding metastases. BJU Int 2005;95:982–6.PubMedCrossRef

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

Bast RC Jr, Ravdin P, Hayes DF, Bates S, Fritsche H Jr, Jessup JM, et al. 2000 update of recommendations for the use of tumor markers in breast and colorectal cancer: clinical practice guidelines of the American Society of Clinical Oncology. J Clin Oncol 2001;19:1865–78.PubMed

Molina R, Barak V, van Dalen A, Duffy MJ, Einarsson R, Gion M, et al. Tumor markers in breast cancer—European Group on Tumor Markers recommendations. Tumour Biol 2005;26:281–93.PubMedCrossRef

Schaller G, Evers K, Papadopoulos S, Ebert A, Buhler H. Current use of HER2 tests. Ann Oncol 2001;12 Suppl 1:S97–100.PubMedCrossRef

Zidan J, Dashkovsky I, Stayerman C, Basher W, Cozacov C, Hadary A. Comparison of HER-2 overexpression in primary breast cancer and metastatic sites and its effect on biological targeting therapy of metastatic disease. Br J Cancer 2005;93:552–6.PubMedCrossRef

Behr TM, Behe M, Wormann B. Trastuzumab and breast cancer. N Engl J Med 2001;345:995–6.PubMedCrossRef

10.

Perik PJ, Lub-De Hooge MN, Gietema JA, van der Graaf WT, de Korte MA, Jonkman S, et al. Indium-111-labeled trastuzumab scintigraphy in patients with human epidermal growth factor receptor 2-positive metastatic breast cancer. J Clin Oncol 2006;24:2276–82.PubMedCrossRef

11.

Batra SK, Jain M, Wittel UA, Chauhan SC, Colcher D. Pharmacokinetics and biodistribution of genetically engineered antibodies. Curr Opin Biotechnol 2002;13:603–8.PubMedCrossRef

12.

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.PubMedCrossRef

13.

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.PubMedCrossRef

14.

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

15.

Orlova A, Nilsson FY, Wikman M, Widstrom C, Stahl 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.PubMed

16.

Steffen AC, Orlova A, Wikman M, Nilsson FY, Stahl S, Adams GP, et al. Affibody-mediated tumour targeting of HER-2 expressing xenografts in mice. Eur J Nucl Med Mol Imaging 2006;33:631–8.PubMedCrossRef

17.

Tolmachev V, Nilsson FY, Widstrom C, Andersson K, Rosik D, Gedda L, et al. ¹¹¹In-benzyl-DTPA-ZHER2:342, an affibody-based conjugate for in vivo imaging of HER2 expression in malignant tumors. J Nucl Med 2006;47:846–53.PubMed

18.

Orlova A, Tolmachev V, Pehrson P, 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 2006;67:2178–86.CrossRef

19.

Baum R, Orlova A, Tolmachev V, Feldwisch J. Receptor PET/CT and SPECT using an Affibody molecule for targeting and molecular imaging of HER2 positive cancer in animal xenografts and human breast cancer patients. J Nucl Med 2006;47 Suppl:108P.

20.

Feldwisch J, Orlova A, Tolmachev V, Baum R. Clinical and pre-clinical application of HER2-specific affibody molecules for diagnosis of recurrent HER2 positive breast cancer by SPECT or PET/CT. Molecular Imaging 2006;5 Suppl:215.

21.

Liu S, Edwards DS. ^99mTc-labeled small peptides as diagnostic radiopharmaceuticals. Chem Rev 1999;99:2235–68.PubMedCrossRef

22.

Liu S, Edwards DS, Barrett JA. ^99mTc labeling of highly potent small peptides. Bioconjug Chem 1997;8:366–621.

23.

Engfeldt T, Tran T, Orlova A, Bruskin A, Widstrom C, Karlström AE, et al. Imaging of HER2-expressing tumours using a synthetic Affibody molecule containing the ^99mTc-chelating mercaptoacetyl-glycyl-glycyl-glycyl (MAG3) sequence. Eur J Nucl Med Mol Imaging 2007;34:722–33.PubMedCrossRef

24.

Zhu Z, Wang Y, Zhang Y, Liu G, Liu N, Rusckowski M, et al. A novel and simplified route to the synthesis of N3S chelators for ^99mTc labeling. Nucl Med Biol 2001;28:703–8.PubMedCrossRef

25.

Decristoforo C, Mather SJ. The influence of chelator on the pharmacokinetics of ^99mTc-labelled peptides. Q J Nucl Med 2002;46:195–205.PubMed

26.

Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol 1982;157:105–32.PubMedCrossRef

27.

Chang F, Rusckowski M, Qu T, Hnatowich DJ. Early results in the irrational design of new bifunctional chelators. Cancer 1997;80:2347–53.PubMedCrossRef

28.

Chang F, Qu T, Rusckowski M, Hnatowich DJ. NHS-MAS3: a bifunctional chelator alternative to NHS-MAG3. Appl Radiat Isot 1999;50:723–32.PubMedCrossRef

29.

Rusckowski M, Qu T, Gupta S, Ley A, Hnatowich DJ. A comparison in monkeys of ^99mTc labeled to a peptide by 4 methods. J Nucl Med 2001;42:1870–7.PubMed

30.

Engfeldt T, Renberg B, Brumer H, Nygren PA, Karlstrom AE. Chemical synthesis of triple-labelled three-helix bundle binding proteins for specific fluorescent detection of unlabelled protein. Chembiochem 2005;6:1043–50.PubMedCrossRef

31.

Tran T, Engfeldt T, Orlova A, Widstrom C, Bruskin A, Tolmachev V, et al. In vivo evaluation of cysteine-based chelators for attachment of ^99mTc to tumor-targeting affibody molecules. Bioconjug Chem 2007;18:549–58.PubMedCrossRef

32.

Behr TM, Gotthardt M, Barth A, Behe M. Imaging tumors with peptide-based radioligands. Q J Nucl Med 2001;45:189–200.PubMed

33.

Qu T, Wang Y, Zhu Z, Rusckowski M, Hnatowich DJ. Different chelators and different peptides together influence the in vitro and mouse in vivo properties of ⁹⁹Tc^m. Nucl Med Commun 2001;22:203–15.PubMedCrossRef

34.

Vanderheyden JL, Liu G, He J, Patel B, Tait JF, Hnatowich DJ. Evaluation of ^99mTc-MAG3-annexin V: influence of the chelate on in vitro and in vivo properties in mice. Nucl Med Biol 2006;33:135–44.PubMedCrossRef

35.

Thakur ML, Marcus CS, Saeed S, Pallela V, Minami C, Diggles L, et al. ^99mTc-labeled vasoactive intestinal peptide analog for rapid localization of tumors in humans. J Nucl Med 2000;41:107–10.PubMed

Titel: 99mTc-chelator engineering to improve tumour targeting properties of a HER2-specific Affibody molecule
verfasst von: Torun Engfeldt
Thuy Tran
Anna Orlova
Charles Widström
Joachim Feldwisch
Lars Abrahmsen
Anders Wennborg
Amelie Eriksson Karlström
Vladimir Tolmachev
Publikationsdatum: 01.11.2007
Verlag: Springer-Verlag
Erschienen in: European Journal of Nuclear Medicine and Molecular Imaging / Ausgabe 11/2007
Print ISSN: 1619-7070
Elektronische ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-007-0474-6

Springer Medizin

Abstract

Purpose

Methods

Results

Conclusion

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 11/2007

PET imaging of acute and chronic inflammation in living mice

Yttrium 90 Bremsstrahlung SPECT/CT scan demonstrating areas of tracer/tumour uptake

In vivo imaging of the endocannabinoid system: a novel window to a central modulatory mechanism in humans

Martin Charron, editor. Practical pediatric PET imaging

An analysis of the physiological FDG uptake in the stomach with the water gastric distention method

The new EANM paediatric dosage card — does it conform to ALARA for PET/CT?