Due to its frequent overexpression in a variety of solid tumors the epidermal growth factor receptor (EGFR) is a well-established target for therapeutic interventions in epithelial cancers. In order to target EGFR in head and neck cancer, we have generated a ribonuclease (RNase) fusion protein comprising a humanized anti-EGFR antibody single-chain Fv fragment (scFv) and Ranpirnase, an RNase from Rana pipiens. Fusion of Ranpirnase to the N-terminus of the scFv via a flexible glycine-serine linker (G4S)3 resulted in very poor cytotoxicity of the fusion protein. As endosomal accumulation and lysosomal degradation have been reported to diminish the antitumor efficacy of ribonuclease or toxin-based immunoagents, we explored a fusion peptide from dengue virus that has been reported to be involved in the endosomal escape of the virus. This peptide was introduced as a linker between Ranpirnase and the scFv moiety. The modified immunoRNase exhibited exceptionally high cytotoxicity toward EGFR-expressing head and neck cell lines without affecting specificity. These results indicate that endosomal entrapment needs to be considered for Ranpirnase-based immunoagents and might be overcome by the use of tailored transduction domains from viral proteins.
%0 Journal Article
%1 Kiesgen2014
%A Kiesgen, Stefan
%A Liebers, Nora
%A Cremer, Martin
%A Arnold, Ulrich
%A Weber, Tobias
%A Keller, Armin
%A Herold-Mende, Christel
%A Dyckhoff, Gerhard
%A Jäger, Dirk
%A Kontermann, Roland E.
%A Arndt, Michaela A.E.
%A Krauss, Jürgen
%A Wu, Anna
%D 2014
%J Protein Engineering, Design and Selection
%K 2014 izi kontermann
%N 10
%P 331--337
%R 10.1093/protein/gzu040
%T A fusogenic dengue virus-derived peptide enhances antitumor efficacy of an antibody-ribonuclease fusion protein targeting the EGF receptor
%U http://www.ncbi.nlm.nih.gov/pubmed/25301960
%V 27
%X Due to its frequent overexpression in a variety of solid tumors the epidermal growth factor receptor (EGFR) is a well-established target for therapeutic interventions in epithelial cancers. In order to target EGFR in head and neck cancer, we have generated a ribonuclease (RNase) fusion protein comprising a humanized anti-EGFR antibody single-chain Fv fragment (scFv) and Ranpirnase, an RNase from Rana pipiens. Fusion of Ranpirnase to the N-terminus of the scFv via a flexible glycine-serine linker (G4S)3 resulted in very poor cytotoxicity of the fusion protein. As endosomal accumulation and lysosomal degradation have been reported to diminish the antitumor efficacy of ribonuclease or toxin-based immunoagents, we explored a fusion peptide from dengue virus that has been reported to be involved in the endosomal escape of the virus. This peptide was introduced as a linker between Ranpirnase and the scFv moiety. The modified immunoRNase exhibited exceptionally high cytotoxicity toward EGFR-expressing head and neck cell lines without affecting specificity. These results indicate that endosomal entrapment needs to be considered for Ranpirnase-based immunoagents and might be overcome by the use of tailored transduction domains from viral proteins.
%@ 1741-0134 (Electronic)$\backslash$r1741-0126 (Linking)
@article{Kiesgen2014,
abstract = {Due to its frequent overexpression in a variety of solid tumors the epidermal growth factor receptor (EGFR) is a well-established target for therapeutic interventions in epithelial cancers. In order to target EGFR in head and neck cancer, we have generated a ribonuclease (RNase) fusion protein comprising a humanized anti-EGFR antibody single-chain Fv fragment (scFv) and Ranpirnase, an RNase from Rana pipiens. Fusion of Ranpirnase to the N-terminus of the scFv via a flexible glycine-serine linker (G4S)3 resulted in very poor cytotoxicity of the fusion protein. As endosomal accumulation and lysosomal degradation have been reported to diminish the antitumor efficacy of ribonuclease or toxin-based immunoagents, we explored a fusion peptide from dengue virus that has been reported to be involved in the endosomal escape of the virus. This peptide was introduced as a linker between Ranpirnase and the scFv moiety. The modified immunoRNase exhibited exceptionally high cytotoxicity toward EGFR-expressing head and neck cell lines without affecting specificity. These results indicate that endosomal entrapment needs to be considered for Ranpirnase-based immunoagents and might be overcome by the use of tailored transduction domains from viral proteins.},
added-at = {2023-06-29T13:07:55.000+0200},
author = {Kiesgen, Stefan and Liebers, Nora and Cremer, Martin and Arnold, Ulrich and Weber, Tobias and Keller, Armin and Herold-Mende, Christel and Dyckhoff, Gerhard and J{\"{a}}ger, Dirk and Kontermann, Roland E. and Arndt, Michaela A.E. and Krauss, J{\"{u}}rgen and Wu, Anna},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2c47072877fdb4a58d9b79243c86d7347/fabian},
doi = {10.1093/protein/gzu040},
interhash = {56f6e127e6af8b08c5f7525b51ca551d},
intrahash = {c47072877fdb4a58d9b79243c86d7347},
isbn = {1741-0134 (Electronic)$\backslash$r1741-0126 (Linking)},
issn = {17410134},
journal = {Protein Engineering, Design and Selection},
keywords = {2014 izi kontermann},
month = oct,
number = 10,
pages = {331--337},
pmid = {25301960},
timestamp = {2023-06-29T13:07:55.000+0200},
title = {{A fusogenic dengue virus-derived peptide enhances antitumor efficacy of an antibody-ribonuclease fusion protein targeting the EGF receptor}},
url = {http://www.ncbi.nlm.nih.gov/pubmed/25301960},
volume = 27,
year = 2014
}
