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The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis

We recently found that integrin αvβ3 binds to fibroblast growth factor (FGF)-αvβ31 (FGF1), and that the integrin-binding defective FGF1 mutant (Arg-50 to glutamic acid, R50E) is defective in signalling and antagonistic to FGF1 signalling. R50E suppressed angiogenesis and tumour growth, suggesting th...

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Autores principales: Mori, Seiji, Hatori, Nobuaki, Kawaguchi, Naomasa, Hamada, Yoshinosuke, Shih, Tsung-Chieh, Wu, Chun-Yi, Lam, Kit S., Matsuura, Nariaki, Yamamoto, Hirofumi, Takada, Yoko K., Takada, Yoshikazu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5482197/
https://www.ncbi.nlm.nih.gov/pubmed/28302677
http://dx.doi.org/10.1042/BSR20170173
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author Mori, Seiji
Hatori, Nobuaki
Kawaguchi, Naomasa
Hamada, Yoshinosuke
Shih, Tsung-Chieh
Wu, Chun-Yi
Lam, Kit S.
Matsuura, Nariaki
Yamamoto, Hirofumi
Takada, Yoko K.
Takada, Yoshikazu
author_facet Mori, Seiji
Hatori, Nobuaki
Kawaguchi, Naomasa
Hamada, Yoshinosuke
Shih, Tsung-Chieh
Wu, Chun-Yi
Lam, Kit S.
Matsuura, Nariaki
Yamamoto, Hirofumi
Takada, Yoko K.
Takada, Yoshikazu
author_sort Mori, Seiji
collection PubMed
description We recently found that integrin αvβ3 binds to fibroblast growth factor (FGF)-αvβ31 (FGF1), and that the integrin-binding defective FGF1 mutant (Arg-50 to glutamic acid, R50E) is defective in signalling and antagonistic to FGF1 signalling. R50E suppressed angiogenesis and tumour growth, suggesting that R50E has potential as a therapeutic. However, FGF1 is unstable, and we had to express R50E in cancer cells for xenograft study, since injected R50E may rapidly disappear from circulation. We studied if we can develop antagonist of more stable FGF2. FGF2 is widely involved in important biological processes such as stem cell proliferation and angiogenesis. Previous studies found that FGF2 bound to αvβ3 and antagonists to αvβ3 suppressed FGF2-induced angiogenesis. However, it is unclear how FGF2 interacts with integrins. Here, we describe that substituting Lys-119/Arg-120 and Lys-125 residues in the predicted integrin-binding interface of FGF2 to glutamic acid (the K119E/R120E and K125E mutations) effectively reduced integrin binding to FGF2. These FGF2 mutants were defective in signalling functions (ERK1/2 activation and DNA synthesis) in NIH3T3 cells. Notably they suppressed, FGF2 signalling induced by WT FGF2 in endothelial cells, suggesting that the FGF2 mutants are antagonists. The FGF2 mutants effectively suppressed tube formation in vitro, sprouting in aorta ring assays ex vivo and angiogenesis in vivo. The positions of amino acids critical for integrin binding are different between FGF1 and FGF2, suggesting that they do not interact with integrins in the same manner. The newly developed FGF2 mutants have potential as anti-angiogenic agents and useful tools for studying the role of integrins in FGF2 signalling.
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spelling pubmed-54821972017-07-07 The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis Mori, Seiji Hatori, Nobuaki Kawaguchi, Naomasa Hamada, Yoshinosuke Shih, Tsung-Chieh Wu, Chun-Yi Lam, Kit S. Matsuura, Nariaki Yamamoto, Hirofumi Takada, Yoko K. Takada, Yoshikazu Biosci Rep Research Articles We recently found that integrin αvβ3 binds to fibroblast growth factor (FGF)-αvβ31 (FGF1), and that the integrin-binding defective FGF1 mutant (Arg-50 to glutamic acid, R50E) is defective in signalling and antagonistic to FGF1 signalling. R50E suppressed angiogenesis and tumour growth, suggesting that R50E has potential as a therapeutic. However, FGF1 is unstable, and we had to express R50E in cancer cells for xenograft study, since injected R50E may rapidly disappear from circulation. We studied if we can develop antagonist of more stable FGF2. FGF2 is widely involved in important biological processes such as stem cell proliferation and angiogenesis. Previous studies found that FGF2 bound to αvβ3 and antagonists to αvβ3 suppressed FGF2-induced angiogenesis. However, it is unclear how FGF2 interacts with integrins. Here, we describe that substituting Lys-119/Arg-120 and Lys-125 residues in the predicted integrin-binding interface of FGF2 to glutamic acid (the K119E/R120E and K125E mutations) effectively reduced integrin binding to FGF2. These FGF2 mutants were defective in signalling functions (ERK1/2 activation and DNA synthesis) in NIH3T3 cells. Notably they suppressed, FGF2 signalling induced by WT FGF2 in endothelial cells, suggesting that the FGF2 mutants are antagonists. The FGF2 mutants effectively suppressed tube formation in vitro, sprouting in aorta ring assays ex vivo and angiogenesis in vivo. The positions of amino acids critical for integrin binding are different between FGF1 and FGF2, suggesting that they do not interact with integrins in the same manner. The newly developed FGF2 mutants have potential as anti-angiogenic agents and useful tools for studying the role of integrins in FGF2 signalling. Portland Press Ltd. 2017-04-10 /pmc/articles/PMC5482197/ /pubmed/28302677 http://dx.doi.org/10.1042/BSR20170173 Text en © 2017 The Author(s). http://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (http://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Articles
Mori, Seiji
Hatori, Nobuaki
Kawaguchi, Naomasa
Hamada, Yoshinosuke
Shih, Tsung-Chieh
Wu, Chun-Yi
Lam, Kit S.
Matsuura, Nariaki
Yamamoto, Hirofumi
Takada, Yoko K.
Takada, Yoshikazu
The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis
title The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis
title_full The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis
title_fullStr The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis
title_full_unstemmed The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis
title_short The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis
title_sort integrin-binding defective fgf2 mutants potently suppress fgf2 signalling and angiogenesis
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5482197/
https://www.ncbi.nlm.nih.gov/pubmed/28302677
http://dx.doi.org/10.1042/BSR20170173
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