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Effect of single-chain antibody targeting of the ligand-binding domain in the anaplastic lymphoma kinase receptor

Oncogene volume 28, pages 3296–3306 (2009)Cite this article

Abstract

The tyrosine kinase receptor anaplastic lymphoma kinase (ALK) and its ligand, the growth factor pleiotrophin (PTN), are highly expressed during the development of the nervous system and have been implicated in the malignant progression of different tumor types. Here, we describe human single-chain variable fragment (scFv) antibodies that target the ligand-binding domain (LBD) in ALK and show the effect in vitro and in vivo. The ALK LBD was used as a bait in a yeast two-hybdrid system to select human scFv from a library with randomized complementarity-determining region 3 domains. Surface plasmon resonance showed high-affinity binding of the selected scFv. The anti-ALK scFv competed for binding of PTN to ALK in intact cells and inhibited PTN-dependent signal transduction through endogenous ALK. Invasion of an intact endothelial cell monolayer by U87MG human glioblastoma cells was inhibited by the anti-ALK scFv. In addition, the growth of established tumor xenografts in mice was reversed after the induction of the conditional expression of the anti-ALK scFv. In archival malignant brain tumors expression levels of ALK and PTN were found elevated and appear correlated with poor patient survival. This suggests a rate-limiting function of the PTN/ALK interaction that may be exploited therapeutically.

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Acknowledgements

We thank Drs Emma T Bowden, Angera H Kuo and Gerald E Stoica (Georgetown University) for help with some of the experiments as well as discussions and suggestions. This work was supported by RO1 CA108440 from the National Institute of Health (to AW).

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  1. Lombardi Cancer Center, Georgetown University, Washington DC, 20057, USA

    D C Stylianou, D P Kodack, R T Henke, J A Toretsky, A T Riegel & A Wellstein

  2. Esbatech AG, Zürich, Switzerland

    A Auf der Maur & S Hohn

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  1. D C Stylianou
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  2. A Auf der Maur
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  3. D P Kodack
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  4. R T Henke
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Correspondence to A Wellstein.

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Stylianou, D., Auf der Maur, A., Kodack, D. et al. Effect of single-chain antibody targeting of the ligand-binding domain in the anaplastic lymphoma kinase receptor. Oncogene 28, 3296–3306 (2009). https://doi.org/10.1038/onc.2009.184

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