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Development of a specific affinity-matured exosite inhibitor to MT1-MMP that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo

The membrane-associated matrix metalloproteinase-14, MT1-MMP, has been implicated in pericellular proteolysis with an important role in cellular invasion of collagenous tissues. It is substantially upregulated in various cancers and rheumatoid arthritis, and has been considered as a potential therap...

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Autores principales: Botkjaer, Kenneth A., Kwok, Hang Fai, Terp, Mikkel G., Karatt-Vellatt, Aneesh, Santamaria, Salvatore, McCafferty, John, Andreasen, Peter A., Itoh, Yoshifumi, Ditzel, Henrik J., Murphy, Gillian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4941350/
https://www.ncbi.nlm.nih.gov/pubmed/26934448
http://dx.doi.org/10.18632/oncotarget.7780
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author Botkjaer, Kenneth A.
Kwok, Hang Fai
Terp, Mikkel G.
Karatt-Vellatt, Aneesh
Santamaria, Salvatore
McCafferty, John
Andreasen, Peter A.
Itoh, Yoshifumi
Ditzel, Henrik J.
Murphy, Gillian
author_facet Botkjaer, Kenneth A.
Kwok, Hang Fai
Terp, Mikkel G.
Karatt-Vellatt, Aneesh
Santamaria, Salvatore
McCafferty, John
Andreasen, Peter A.
Itoh, Yoshifumi
Ditzel, Henrik J.
Murphy, Gillian
author_sort Botkjaer, Kenneth A.
collection PubMed
description The membrane-associated matrix metalloproteinase-14, MT1-MMP, has been implicated in pericellular proteolysis with an important role in cellular invasion of collagenous tissues. It is substantially upregulated in various cancers and rheumatoid arthritis, and has been considered as a potential therapeutic target. Here, we report the identification of antibody fragments to MT1-MMP that potently and specifically inhibit its cell surface functions. Lead antibody clones displayed inhibitory activity towards pro-MMP-2 activation, collagen-film degradation and gelatin-film degradation, and were shown to bind to the MT1-MMP catalytic domain outside the active site cleft, inhibiting binding to triple helical collagen. Affinity maturation using CDR3 randomization created a second generation of antibody fragments with dissociation constants down to 0.11 nM, corresponding to an improved affinity of 332-fold with the ability to interfere with cell-surface MT1-MMP functions, displaying IC(50) values down to 5 nM. Importantly, the new inhibitors were able to inhibit collagen invasion by tumor-cells in vitro and in vivo primary tumor growth and metastasis of MDA-MB-231 cells in a mouse orthotopic xenograft model. Herein is the first demonstration that an inhibitory antibody targeting sites outside the catalytic cleft of MT1-MMP can effectively abrogate its in vivo activity during tumorigenesis and metastasis.
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spelling pubmed-49413502016-07-19 Development of a specific affinity-matured exosite inhibitor to MT1-MMP that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo Botkjaer, Kenneth A. Kwok, Hang Fai Terp, Mikkel G. Karatt-Vellatt, Aneesh Santamaria, Salvatore McCafferty, John Andreasen, Peter A. Itoh, Yoshifumi Ditzel, Henrik J. Murphy, Gillian Oncotarget Research Paper The membrane-associated matrix metalloproteinase-14, MT1-MMP, has been implicated in pericellular proteolysis with an important role in cellular invasion of collagenous tissues. It is substantially upregulated in various cancers and rheumatoid arthritis, and has been considered as a potential therapeutic target. Here, we report the identification of antibody fragments to MT1-MMP that potently and specifically inhibit its cell surface functions. Lead antibody clones displayed inhibitory activity towards pro-MMP-2 activation, collagen-film degradation and gelatin-film degradation, and were shown to bind to the MT1-MMP catalytic domain outside the active site cleft, inhibiting binding to triple helical collagen. Affinity maturation using CDR3 randomization created a second generation of antibody fragments with dissociation constants down to 0.11 nM, corresponding to an improved affinity of 332-fold with the ability to interfere with cell-surface MT1-MMP functions, displaying IC(50) values down to 5 nM. Importantly, the new inhibitors were able to inhibit collagen invasion by tumor-cells in vitro and in vivo primary tumor growth and metastasis of MDA-MB-231 cells in a mouse orthotopic xenograft model. Herein is the first demonstration that an inhibitory antibody targeting sites outside the catalytic cleft of MT1-MMP can effectively abrogate its in vivo activity during tumorigenesis and metastasis. Impact Journals LLC 2016-02-27 /pmc/articles/PMC4941350/ /pubmed/26934448 http://dx.doi.org/10.18632/oncotarget.7780 Text en Copyright: © 2016 Botkjaer et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Paper
Botkjaer, Kenneth A.
Kwok, Hang Fai
Terp, Mikkel G.
Karatt-Vellatt, Aneesh
Santamaria, Salvatore
McCafferty, John
Andreasen, Peter A.
Itoh, Yoshifumi
Ditzel, Henrik J.
Murphy, Gillian
Development of a specific affinity-matured exosite inhibitor to MT1-MMP that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo
title Development of a specific affinity-matured exosite inhibitor to MT1-MMP that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo
title_full Development of a specific affinity-matured exosite inhibitor to MT1-MMP that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo
title_fullStr Development of a specific affinity-matured exosite inhibitor to MT1-MMP that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo
title_full_unstemmed Development of a specific affinity-matured exosite inhibitor to MT1-MMP that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo
title_short Development of a specific affinity-matured exosite inhibitor to MT1-MMP that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo
title_sort development of a specific affinity-matured exosite inhibitor to mt1-mmp that efficiently inhibits tumor cell invasion in vitro and metastasis in vivo
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4941350/
https://www.ncbi.nlm.nih.gov/pubmed/26934448
http://dx.doi.org/10.18632/oncotarget.7780
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