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Identification of a novel antagonist of the ErbB1 receptor capable of inhibiting migration of human glioblastoma cells

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Abstract

Background

Receptors of the ErbB family are involved in the development of various cancers, and the inhibition of these receptors represents an attractive therapeutic concept. Upon ligand binding, ErbB receptors become activated as homo- or heterodimers, leading to the activation of downstream signaling cascades that result in the facilitation of cell proliferation and migration. A region of the extracellular part of the receptor, termed the ‘dimerization arm’, is important for the formation of receptor dimers and represents an attractive target for the design of ErbB inhibitors.

Methods

An ErbB1 targeting peptide, termed Herfin-1, was designed based on a model of the tertiary structure of the EGF-EGFR ternary complex. The binding kinetics of this peptide were determined employing surface plasmon resonance analyses. ErbB1-4 expression and phosphorylation in human glioblastoma cell lines U87 and U118 were determined by Western blotting using specific antibodies. Cell proliferation was determined by MTS staining. Cell migration was examined using a Chemotaxis Migration Kit. Neurite outgrowth from primary cerebellar granule neurons was evaluated by fluorescence microscopy and image processing.

Results

The present study shows that Herfin-1 functions as an ErbB1 antagonist. It binds to the extracellular domain of ErbB1 with a KD value of 361 nM. In U87 and U118 cells, both expressing high levels of ErbB1, Herfin-1 inhibits EGF-induced ErbB1 phosphorylation and cell migration. Additionally, Herfin-1 was found to increase neurite outgrowth in cerebellar granule neurons, likely through the inhibition of a sustained weak ErbB1 activation.

Conclusions

Targeting the ErbB1 receptor dimerization interface is a promising strategy to inhibit receptor activation in ErbB1-expressing glioma cells.

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Acknowledgments

Help from the Novo Nordisk Foundation Center for Protein Research (Copenhagen, Denmark) with the binding studies performed using the Biacore T200 instrument is greatly appreciated.

Funding

This work was supported by the Lundbeck Foundation (R13-A1398 to V.B.) and the Danish Research Council (09–065919 to V.B., 09–071033 to E.B.).

Conflict of interest

There is no conflict of interest for all authors

Author information

Authors and Affiliations

  1. Department of Neuroscience and Pharmacology, Protein Laboratory, University of Copenhagen, Blegdamsvej 3, 2200, Copenhagen, Denmark

    Mikkel Staberg, Christian Riemer, Ruodan Xu, Oksana Dmytriyeva, Elisabeth Bock & Vladimir Berezin

  2. The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Blegdamsvej 3, 2200, Copenhagen, Denmark

    Mikkel Staberg

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  1. Mikkel Staberg
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  2. Christian Riemer
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Correspondence to Mikkel Staberg.

Electronic supplementary material

Supplementary Fig. 1

Schematic presentation of the Herfin-1 peptide tetramer (JPEG 15 kb)

High resolution image (TIFF 9538 kb)

Supplementary Fig. 2

Sequence alignment of the extended part of the C-terminal (Herfin motifs) and N-terminal (Herfin counter-motifs involved in hydrophobic interactions with the Herfin motif) regions of the dimerization arm of four ErbB receptors (ErbB1, ErbB2, ErbB3, and ErbB4; UniProtKB/Swiss-Prot accession no. P00533, P04626, P21860, and Q15303, respectively) (JPEG 47 kb)

High resolution image (TIFF 14290 kb)

Supplementary Fig. 3

Sequence alignment of the N-terminal region of the ErbB1 dimerization arm and N-terminal region of E-cadherin extracellular domain 3 (UniProtKB/Swiss-Prot accession no. P00533 and P09803, respectively). The advanced BLAST program was used (http://www.ch.embnet.org/software/bBLAST.html; accessed June 6, 2012) (JPEG 11 kb)

High resolution image (TIFF 9493 kb)

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Staberg, M., Riemer, C., Xu, R. et al. Identification of a novel antagonist of the ErbB1 receptor capable of inhibiting migration of human glioblastoma cells. Cell Oncol. 36, 201–211 (2013). https://doi.org/10.1007/s13402-013-0128-6

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