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Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates

We recently reported on a potent synthetic agent, 135H11, that selectively targets the receptor tyrosine kinase, EphA2. While 135H11 possesses a relatively high binding affinity for the ligand-binding domain of EphA2 (Kd~130 nM), receptor activation in the cell required the synthesis of dimeric vers...

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Autores principales: Udompholkul, Parima, Baggio, Carlo, Gambini, Luca, Sun, Yu, Zhao, Ming, Hoffman, Robert M., Pellecchia, Maurizio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8235110/
https://www.ncbi.nlm.nih.gov/pubmed/34204178
http://dx.doi.org/10.3390/molecules26123687
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author Udompholkul, Parima
Baggio, Carlo
Gambini, Luca
Sun, Yu
Zhao, Ming
Hoffman, Robert M.
Pellecchia, Maurizio
author_facet Udompholkul, Parima
Baggio, Carlo
Gambini, Luca
Sun, Yu
Zhao, Ming
Hoffman, Robert M.
Pellecchia, Maurizio
author_sort Udompholkul, Parima
collection PubMed
description We recently reported on a potent synthetic agent, 135H11, that selectively targets the receptor tyrosine kinase, EphA2. While 135H11 possesses a relatively high binding affinity for the ligand-binding domain of EphA2 (Kd~130 nM), receptor activation in the cell required the synthesis of dimeric versions of such agent (namely 135H12). This was expected given that the natural ephrin ligands also need to be dimerized or clustered to elicit agonistic activity in cell. In the present report we investigated whether the agonistic activity of 135H11 could be enhanced by biotin conjugation followed by complex formation with streptavidin. Therefore, we measured the agonistic EphA2 activity of 135H11-biotin (147B5) at various agent/streptavidin ratios, side by side with 135H12, and a scrambled version of 147B5 in pancreatic- and breast-cancer cell lines. The (147B5)(n)-streptavidin complexes (when n = 2, 3, 4, but not when n = 1) induced a strong receptor degradation effect in both cell lines compared to 135H12 or the (scrambled-147B5)(4)-streptavidin complex as a control, indicating that multimerization of the targeting agent resulted in an increased ability to cause receptor clustering and internalization. Subsequently, we prepared an Alexa-Fluor-streptavidin conjugate to demonstrate that (147B5)(4)-AF-streptavidin, but not the scrambled equivalent complex, concentrates in pancreatic and breast cancers in orthotopic nude-mouse models. Hence, we conclude that these novel targeting agents, with proper derivatization with imaging reagents or chemotherapy, can be used as diagnostics, and/or to deliver chemotherapy selectively to EphA2-expressing tumors.
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spelling pubmed-82351102021-06-27 Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates Udompholkul, Parima Baggio, Carlo Gambini, Luca Sun, Yu Zhao, Ming Hoffman, Robert M. Pellecchia, Maurizio Molecules Article We recently reported on a potent synthetic agent, 135H11, that selectively targets the receptor tyrosine kinase, EphA2. While 135H11 possesses a relatively high binding affinity for the ligand-binding domain of EphA2 (Kd~130 nM), receptor activation in the cell required the synthesis of dimeric versions of such agent (namely 135H12). This was expected given that the natural ephrin ligands also need to be dimerized or clustered to elicit agonistic activity in cell. In the present report we investigated whether the agonistic activity of 135H11 could be enhanced by biotin conjugation followed by complex formation with streptavidin. Therefore, we measured the agonistic EphA2 activity of 135H11-biotin (147B5) at various agent/streptavidin ratios, side by side with 135H12, and a scrambled version of 147B5 in pancreatic- and breast-cancer cell lines. The (147B5)(n)-streptavidin complexes (when n = 2, 3, 4, but not when n = 1) induced a strong receptor degradation effect in both cell lines compared to 135H12 or the (scrambled-147B5)(4)-streptavidin complex as a control, indicating that multimerization of the targeting agent resulted in an increased ability to cause receptor clustering and internalization. Subsequently, we prepared an Alexa-Fluor-streptavidin conjugate to demonstrate that (147B5)(4)-AF-streptavidin, but not the scrambled equivalent complex, concentrates in pancreatic and breast cancers in orthotopic nude-mouse models. Hence, we conclude that these novel targeting agents, with proper derivatization with imaging reagents or chemotherapy, can be used as diagnostics, and/or to deliver chemotherapy selectively to EphA2-expressing tumors. MDPI 2021-06-17 /pmc/articles/PMC8235110/ /pubmed/34204178 http://dx.doi.org/10.3390/molecules26123687 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Udompholkul, Parima
Baggio, Carlo
Gambini, Luca
Sun, Yu
Zhao, Ming
Hoffman, Robert M.
Pellecchia, Maurizio
Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates
title Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates
title_full Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates
title_fullStr Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates
title_full_unstemmed Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates
title_short Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates
title_sort effective tumor targeting by epha2-agonist-biotin-streptavidin conjugates
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8235110/
https://www.ncbi.nlm.nih.gov/pubmed/34204178
http://dx.doi.org/10.3390/molecules26123687
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