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Article

Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain

  • Authors:
    • Hao Liu
    • Johan Seijsing
    • Fredrik Y. Frejd
    • Vladimir Tolmachev
    • Torbjörn Gräslund
  • View Affiliations / Copyright

    Affiliations: School of Biotechnology, Division of Protein Technology, KTH - Royal Institute of Technology, Stockholm, Sweden, Affibody AB, Solna, Sweden, Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
  • Pages: 601-609
    |
    Published online on: June 4, 2015
       https://doi.org/10.3892/ijo.2015.3027
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Abstract

Development of cancer treatment regimens including immunotoxins is partly hampered by their immunogenicity. Recently, deimmunized versions of toxins have been described, potentially being better suited for translation to the clinic. In this study, a recombinant tripartite fusion toxin consisting of a deimmunized version of exotoxin A from Pseudomonas aeruginosa (PE38) genetically fused to an affibody molecule specifically interacting with the human epidermal growth factor receptor 2 (HER2), and also an albumin binding domain (ABD) for half-life extension, has been produced and characterized in terms of functionality of the three moieties. Biosensor based assays showed that the fusion toxin was able to interact with human and mouse serum albumin, but not with bovine serum albumin and that it interacted with HER2 (KD=5 nM). Interestingly, a complex of the fusion toxin and human serum albumin also interacted with HER2 but with a somewhat weaker affinity (KD=12 nM). The IC50-values of the fusion toxin ranged from 6 to 300 pM on SKOV-3, SKBR-3 and A549 cells and was lower for cells with higher surface densities of HER2. The fusion toxin was found specific for HER2 as shown by blocking available HER2 receptors with free affibody molecule before subjecting the cells to the toxin. Analysis of contact time showed that 10 min was sufficient to kill 50% of the cells. In conclusion, all three regions of the fusion toxin were found to be functional.
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View References

1 

Kreitman RJ: Recombinant immunotoxins containing truncated bacterial toxins for the treatment of hematologic malignancies. BioDrugs. 23:1–13. 2009. View Article : Google Scholar : PubMed/NCBI

2 

Alewine C, Hassan R and Pastan I: Advances in anticancer immunotoxin therapy. Oncologist. 20:176–185. 2015. View Article : Google Scholar : PubMed/NCBI

3 

Martin-Killias P, Stefan N, Rothschild S, Plückthun A and Zangemeister-Wittke U and Zangemeister-Wittke U: A novel fusion toxin derived from an EpCAM-specific designed ankyrin repeat protein has potent antitumor activity. Clin Cancer Res. 17:100–110. 2011. View Article : Google Scholar

4 

Zielinski R, Lyakhov I, Jacobs A, Chertov O, Kramer-Marek G, Francella N, Stephen A, Fisher R, Blumenthal R and Capala J: Affitoxin - a novel recombinant, HER2-specific, anticancer agent for targeted therapy of HER2-positive tumors. J Immunother. 32:817–825. 2009. View Article : Google Scholar : PubMed/NCBI

5 

Zielinski R, Lyakhov I, Hassan M, Kuban M, Shafer-Weaver K, Gandjbakhche A and Capala J: HER2-affitoxin: A potent therapeutic agent for the treatment of HER2-overexpressing tumors. Clin Cancer Res. 17:5071–5081. 2011. View Article : Google Scholar : PubMed/NCBI

6 

Mazor R, Vassall AN, Eberle JA, Beers R, Weldon JE, Venzon DJ, Tsang KY, Benhar I and Pastan I: Identification and elimination of an immunodominant T-cell epitope in recombinant immunotoxins based on Pseudomonas exotoxin A. Proc Natl Acad Sci USA. 109:E3597–E3603. 2012. View Article : Google Scholar : PubMed/NCBI

7 

Onda M, Beers R, Xiang L, Nagata S, Wang Q-C and Pastan I: An immunotoxin with greatly reduced immunogenicity by identification and removal of B cell epitopes. Proc Natl Acad Sci USA. 105:11311–11316. 2008. View Article : Google Scholar : PubMed/NCBI

8 

Liu W, Onda M, Lee B, Kreitman RJ, Hassan R, Xiang L and Pastan I: Recombinant immunotoxin engineered for low immunogenicity and antigenicity by identifying and silencing human B-cell epitopes. Proc Natl Acad Sci USA. 109:11782–11787. 2012. View Article : Google Scholar : PubMed/NCBI

9 

Onda M, Beers R, Xiang L, Lee B, Weldon JE, Kreitman RJ and Pastan I: Recombinant immunotoxin against B-cell malignancies with no immunogenicity in mice by removal of B-cell epitopes. Proc Natl Acad Sci USA. 108:5742–5747. 2011. View Article : Google Scholar : PubMed/NCBI

10 

King C, Garza EN, Mazor R, Linehan JL, Pastan I, Pepper M and Baker D: Removing T-cell epitopes with computational protein design. Proc Natl Acad Sci USA. 111:8577–8582. 2014. View Article : Google Scholar : PubMed/NCBI

11 

Carlsson J, Nordgren H, Sjöström J, Wester K, Villman K, Bengtsson NO, Ostenstad B, Lundqvist H and Blomqvist C: HER2 expression in breast cancer primary tumours and corresponding metastases. Original data and literature review. Br J Cancer. 90:2344–2348. 2004.PubMed/NCBI

12 

Ménard S, Casalini P, Campiglio M, Pupa S, Agresti R and Tagliabue E: HER2 overexpression in various tumor types, focussing on its relationship to the development of invasive breast cancer. Ann Oncol. 12(Suppl 1): S15–S19. 2001. View Article : Google Scholar : PubMed/NCBI

13 

Ahmed S, Sami A and Xiang J: HER2-directed therapy: Current treatment options for HER2-positive breast cancer. Breast Cancer. 22:101–116. 2015. View Article : Google Scholar : PubMed/NCBI

14 

Amiri-Kordestani L, Blumenthal GM, Xu QC, Zhang L, Tang SW, Ha L, Weinberg WC, Chi B, Candau-Chacon R, Hughes P, et al: FDA approval: Ado-trastuzumab emtansine for the treatment of patients with HER2-positive metastatic breast cancer. Clin Cancer Res. 20:4436–4441. 2014. View Article : Google Scholar : PubMed/NCBI

15 

Löfblom J, Feldwisch J, Tolmachev V, Carlsson J, Ståhl S and Frejd FY: Affibody molecules: Engineered proteins for therapeutic, diagnostic and biotechnological applications. FEBS Lett. 584:2670–2680. 2010. View Article : Google Scholar : PubMed/NCBI

16 

Li J, Lundberg E, Vernet E, Larsson B, Höidén-Guthenberg I and Gräslund T: Selection of affibody molecules to the ligand-binding site of the insulin-like growth factor-1 receptor. Biotechnol Appl Biochem. 55:99–109. 2010.PubMed/NCBI

17 

Friedman M, Nordberg E, Höidén-Guthenberg I, Brismar H, Adams GP, Nilsson FY, Carlsson J and Ståhl S: Phage display selection of Affibody molecules with specific binding to the extracellular domain of the epidermal growth factor receptor. Protein Eng Des Sel. 20:189–199. 2007. View Article : Google Scholar : PubMed/NCBI

18 

Orlova A, Magnusson M, Eriksson TLJ, Nilsson M, Larsson B, Höidén-Guthenberg I, Widström C, Carlsson J, Tolmachev V, Ståhl S, et al: Tumor imaging using a picomolar affinity HER2 binding affibody molecule. Cancer Res. 66:4339–4348. 2006. View Article : Google Scholar : PubMed/NCBI

19 

Kronqvist N, Malm M, Göstring L, Gunneriusson E, Nilsson M, Höidén Guthenberg I, Gedda L, Frejd FY, Ståhl S and Löfblom J: Combining phage and staphylococcal surface display for generation of ErbB3-specific Affibody molecules. Protein Eng Des Sel. 24:385–396. 2011. View Article : Google Scholar

20 

Feldwisch J, Tolmachev V, Lendel C, Herne N, Sjöberg A, Larsson B, Rosik D, Lindqvist E, Fant G, Höidén-Guthenberg I, et al: Design of an optimized scaffold for affibody molecules. J Mol Biol. 398:232–247. 2010. View Article : Google Scholar : PubMed/NCBI

21 

Sörensen J, Sandberg D, Sandström M, Wennborg A, Feldwisch J, Tolmachev V, Åström G, Lubberink M, Garske-Román U, Carlsson J, et al: First-in-human molecular imaging of HER2 expression in breast cancer metastases using the 111In-ABY-025 affibody molecule. J Nucl Med. 55:730–735. 2014. View Article : Google Scholar : PubMed/NCBI

22 

Makrides SC, Nygren PA, Andrews B, Ford PJ, Evans KS, Hayman EG, Adari H, Uhlén M and Toth CA: Extended in vivo half-life of human soluble complement receptor type 1 fused to a serum albumin-binding receptor. J Pharmacol Exp Ther. 277:534–542. 1996.PubMed/NCBI

23 

Jonsson A, Dogan J, Herne N, Abrahmsén L and Nygren P-A: Engineering of a femtomolar affinity binding protein to human serum albumin. Protein Eng Des Sel. 21:515–527. 2008. View Article : Google Scholar : PubMed/NCBI

24 

Orlova A, Jonsson A, Rosik D, Lundqvist H, Lindborg M, Abrahmsen L, Ekblad C, Frejd FY and Tolmachev V: Site-specific radiometal labeling and improved biodistribution using ABY-027, a novel HER2-targeting affibody molecule-albumin-binding domain fusion protein. J Nucl Med. 54:961–968. 2013. View Article : Google Scholar : PubMed/NCBI

25 

Tolmachev V, Orlova A, Pehrson R, Galli J, Baastrup B, Andersson K, Sandström M, Rosik D, Carlsson J, Lundqvist H, et al: Radionuclide therapy of HER2-positive microxenografts using a 177Lu-labeled HER2-specific Affibody molecule. Cancer Res. 67:2773–2782. 2007. View Article : Google Scholar : PubMed/NCBI

26 

Hopp J, Hornig N, Zettlitz KA, Schwarz A, Fuss N, Müller D and Kontermann RE: The effects of affinity and valency of an albumin-binding domain (ABD) on the half-life of a single-chain diabody-ABD fusion protein. Protein Eng Des Sel. 23:827–834. 2010. View Article : Google Scholar : PubMed/NCBI

27 

Andersen JT, Pehrson R, Tolmachev V, Daba MB, Abrahmsén L and Ekblad C: Extending half-life by indirect targeting of the neonatal Fc receptor (FcRn) using a minimal albumin binding domain. J Biol Chem. 286:5234–5241. 2011. View Article : Google Scholar :

28 

Gunneriusson E, Nord K, Uhlén M and Nygren P: Affinity maturation of a Taq DNA polymerase specific affibody by helix shuffling. Protein Eng. 12:873–878. 1999. View Article : Google Scholar : PubMed/NCBI

29 

Sandersjöö L, Jonsson A and Löfblom J: A new prodrug form of Affibody molecules (pro-Affibody) is selectively activated by cancer-associated proteases. Cell Mol Life Sci. 72:1405–1415. 2015. View Article : Google Scholar

30 

Tolmachev V, Hofström C, Malmberg J, Ahlgren S, Hosseinimehr SJ, Sandström M, Abrahmsén L, Orlova A and Gräslund T: HEHEHE-tagged affibody molecule may be purified by IMAC, is conveniently labeled with [99mTc(CO)3]+, and shows improved biodistribution with reduced hepatic radioactivity accumulation. Bioconjug Chem. 21:2013–2022. 2010. View Article : Google Scholar : PubMed/NCBI

31 

Hassan R, Bullock S, Premkumar A, Kreitman RJ, Kindler H, Willingham MC and Pastan I: Phase I study of SS1P, a recombinant anti-mesothelin immunotoxin given as a bolus I.V. infusion to patients with mesothelin-expressing mesothelioma, ovarian, and pancreatic cancers. Clin Cancer Res. 13:5144–5149. 2007. View Article : Google Scholar : PubMed/NCBI

32 

Kuan CT, Pai LH and Pastan I: Immunotoxins containing Pseudomonas exotoxin that target LeY damage human endothelial cells in an antibody-specific mode: Relevance to vascular leak syndrome. Clin Cancer Res. 1:1589–1594. 1995.PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Liu H, Seijsing J, Frejd FY, Tolmachev V and Gräslund T: Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain. Int J Oncol 47: 601-609, 2015.
APA
Liu, H., Seijsing, J., Frejd, F.Y., Tolmachev, V., & Gräslund, T. (2015). Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain. International Journal of Oncology, 47, 601-609. https://doi.org/10.3892/ijo.2015.3027
MLA
Liu, H., Seijsing, J., Frejd, F. Y., Tolmachev, V., Gräslund, T."Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain". International Journal of Oncology 47.2 (2015): 601-609.
Chicago
Liu, H., Seijsing, J., Frejd, F. Y., Tolmachev, V., Gräslund, T."Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain". International Journal of Oncology 47, no. 2 (2015): 601-609. https://doi.org/10.3892/ijo.2015.3027
Copy and paste a formatted citation
x
Spandidos Publications style
Liu H, Seijsing J, Frejd FY, Tolmachev V and Gräslund T: Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain. Int J Oncol 47: 601-609, 2015.
APA
Liu, H., Seijsing, J., Frejd, F.Y., Tolmachev, V., & Gräslund, T. (2015). Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain. International Journal of Oncology, 47, 601-609. https://doi.org/10.3892/ijo.2015.3027
MLA
Liu, H., Seijsing, J., Frejd, F. Y., Tolmachev, V., Gräslund, T."Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain". International Journal of Oncology 47.2 (2015): 601-609.
Chicago
Liu, H., Seijsing, J., Frejd, F. Y., Tolmachev, V., Gräslund, T."Target-specific cytotoxic effects on HER2-expressing cells by the tripartite fusion toxin ZHER2:2891-ABD-PE38X8, including a targeting affibody molecule and a half-life extension domain". International Journal of Oncology 47, no. 2 (2015): 601-609. https://doi.org/10.3892/ijo.2015.3027
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