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Targeting the Tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis
BACKGROUND: Increased activity of the receptor tyrosine kinase Tie2 has been implicated in the promotion of pathological angiogenesis. This activity is mainly mediated through angiopoietin (Ang)1- and Ang2-dependent activation of integrins by Tie2, rendering the Ang/Tie2/integrin axis an attractive...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6097439/ https://www.ncbi.nlm.nih.gov/pubmed/30119679 http://dx.doi.org/10.1186/s12915-018-0557-9 |
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author | Shlamkovich, Tomer Aharon, Lidan Koslawsky, Dana Einav, Yulia Papo, Niv |
author_facet | Shlamkovich, Tomer Aharon, Lidan Koslawsky, Dana Einav, Yulia Papo, Niv |
author_sort | Shlamkovich, Tomer |
collection | PubMed |
description | BACKGROUND: Increased activity of the receptor tyrosine kinase Tie2 has been implicated in the promotion of pathological angiogenesis. This activity is mainly mediated through angiopoietin (Ang)1- and Ang2-dependent activation of integrins by Tie2, rendering the Ang/Tie2/integrin axis an attractive putative target for cancer therapeutics. RESULTS: To target this axis, we developed single domain, non-immunoglobulin high-affinity bi-specific protein inhibitors against both Tie2 and α(v)β(3) integrin. We have previously engineered the Ang2-binding domain of Tie2 (Ang2-BD) as a Tie2 inhibitor. Here, we engineered an exposed loop in Ang2-BD to generate variants that include an integrin-binding Arg–Gly–Asp (RGD) motif and used flow cytometry screening of a yeast-displayed Ang2-BD RGD loop library to identify the integrin antagonists. The bi-specific antagonists targeting both Tie2 and α(v)β(3) integrin inhibited adhesion and proliferation of endothelial cells cultured together with the α(v)β(3) integrin ligand vitronectin, as well as endothelial cell invasion and tube formation. The bi-specific reagents inhibited downstream signaling by Tie2 intracellularly in response to its agonist Ang1 more effectively than the wild-type Ang2 BD that binds Tie2 alone. CONCLUSIONS: Collectively, this study—the first to describe inhibitors targeting all the known functions resulting from Tie2/integrin α(v)β(3) cross-talk—has created new tools for studying Tie2- and integrin α(v)β(3)-dependent molecular pathways and provides the basis for the rational and combinatorial engineering of ligand–Tie2 and ligand–integrin α(v)β(3) receptor interactions. Given the roles of these pathways in cancer angiogenesis and metastasis, this proof of principle study paves the route to create novel Tie2/integrin α(v)β(3)-targeting proteins for clinical use as imaging and therapeutic agents. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12915-018-0557-9) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6097439 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-60974392018-08-20 Targeting the Tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis Shlamkovich, Tomer Aharon, Lidan Koslawsky, Dana Einav, Yulia Papo, Niv BMC Biol Research Article BACKGROUND: Increased activity of the receptor tyrosine kinase Tie2 has been implicated in the promotion of pathological angiogenesis. This activity is mainly mediated through angiopoietin (Ang)1- and Ang2-dependent activation of integrins by Tie2, rendering the Ang/Tie2/integrin axis an attractive putative target for cancer therapeutics. RESULTS: To target this axis, we developed single domain, non-immunoglobulin high-affinity bi-specific protein inhibitors against both Tie2 and α(v)β(3) integrin. We have previously engineered the Ang2-binding domain of Tie2 (Ang2-BD) as a Tie2 inhibitor. Here, we engineered an exposed loop in Ang2-BD to generate variants that include an integrin-binding Arg–Gly–Asp (RGD) motif and used flow cytometry screening of a yeast-displayed Ang2-BD RGD loop library to identify the integrin antagonists. The bi-specific antagonists targeting both Tie2 and α(v)β(3) integrin inhibited adhesion and proliferation of endothelial cells cultured together with the α(v)β(3) integrin ligand vitronectin, as well as endothelial cell invasion and tube formation. The bi-specific reagents inhibited downstream signaling by Tie2 intracellularly in response to its agonist Ang1 more effectively than the wild-type Ang2 BD that binds Tie2 alone. CONCLUSIONS: Collectively, this study—the first to describe inhibitors targeting all the known functions resulting from Tie2/integrin α(v)β(3) cross-talk—has created new tools for studying Tie2- and integrin α(v)β(3)-dependent molecular pathways and provides the basis for the rational and combinatorial engineering of ligand–Tie2 and ligand–integrin α(v)β(3) receptor interactions. Given the roles of these pathways in cancer angiogenesis and metastasis, this proof of principle study paves the route to create novel Tie2/integrin α(v)β(3)-targeting proteins for clinical use as imaging and therapeutic agents. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12915-018-0557-9) contains supplementary material, which is available to authorized users. BioMed Central 2018-08-17 /pmc/articles/PMC6097439/ /pubmed/30119679 http://dx.doi.org/10.1186/s12915-018-0557-9 Text en © Papo et al. 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Shlamkovich, Tomer Aharon, Lidan Koslawsky, Dana Einav, Yulia Papo, Niv Targeting the Tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis |
title | Targeting the Tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis |
title_full | Targeting the Tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis |
title_fullStr | Targeting the Tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis |
title_full_unstemmed | Targeting the Tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis |
title_short | Targeting the Tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis |
title_sort | targeting the tie2–α(v)β(3) integrin axis with bi-specific reagents for the inhibition of angiogenesis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6097439/ https://www.ncbi.nlm.nih.gov/pubmed/30119679 http://dx.doi.org/10.1186/s12915-018-0557-9 |
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