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Identification and Validation of an Aspergillus nidulans Secondary Metabolite Derivative as an Inhibitor of the Musashi-RNA Interaction
RNA-binding protein Musashi-1 (MSI1) is a key regulator of several stem cell populations. MSI1 is involved in tumor proliferation and maintenance, and it regulates target mRNAs at the translational level. The known mRNA targets of MSI1 include Numb, APC, and P21(WAF-1), key regulators of Notch/Wnt s...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7463734/ https://www.ncbi.nlm.nih.gov/pubmed/32784494 http://dx.doi.org/10.3390/cancers12082221 |
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author | Lan, Lan Liu, Jiajun Xing, Minli Smith, Amber R. Wang, Jinan Wu, Xiaoqing Appelman, Carl Li, Ke Roy, Anuradha Gowthaman, Ragul Karanicolas, John Somoza, Amber D. Wang, Clay C. C. Miao, Yinglong De Guzman, Roberto Oakley, Berl R. Neufeld, Kristi L. Xu, Liang |
author_facet | Lan, Lan Liu, Jiajun Xing, Minli Smith, Amber R. Wang, Jinan Wu, Xiaoqing Appelman, Carl Li, Ke Roy, Anuradha Gowthaman, Ragul Karanicolas, John Somoza, Amber D. Wang, Clay C. C. Miao, Yinglong De Guzman, Roberto Oakley, Berl R. Neufeld, Kristi L. Xu, Liang |
author_sort | Lan, Lan |
collection | PubMed |
description | RNA-binding protein Musashi-1 (MSI1) is a key regulator of several stem cell populations. MSI1 is involved in tumor proliferation and maintenance, and it regulates target mRNAs at the translational level. The known mRNA targets of MSI1 include Numb, APC, and P21(WAF-1), key regulators of Notch/Wnt signaling and cell cycle progression, respectively. In this study, we aim to identify small molecule inhibitors of MSI1–mRNA interactions, which could block the growth of cancer cells with high levels of MSI1. Using a fluorescence polarization (FP) assay, we screened small molecules from several chemical libraries for those that disrupt the binding of MSI1 to its consensus RNA. One cluster of hit compounds is the derivatives of secondary metabolites from Aspergillus nidulans. One of the top hits, Aza-9, from this cluster was further validated by surface plasmon resonance and nuclear magnetic resonance spectroscopy, which demonstrated that Aza-9 binds directly to MSI1, and the binding is at the RNA binding pocket. We also show that Aza-9 binds to Musashi-2 (MSI2) as well. To test whether Aza-9 has anti-cancer potential, we used liposomes to facilitate Aza-9 cellular uptake. Aza-9-liposome inhibits proliferation, induces apoptosis and autophagy, and down-regulates Notch and Wnt signaling in colon cancer cell lines. In conclusion, we identified a series of potential lead compounds for inhibiting MSI1/2 function, while establishing a framework for identifying small molecule inhibitors of RNA binding proteins using FP-based screening methodology. |
format | Online Article Text |
id | pubmed-7463734 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-74637342020-09-02 Identification and Validation of an Aspergillus nidulans Secondary Metabolite Derivative as an Inhibitor of the Musashi-RNA Interaction Lan, Lan Liu, Jiajun Xing, Minli Smith, Amber R. Wang, Jinan Wu, Xiaoqing Appelman, Carl Li, Ke Roy, Anuradha Gowthaman, Ragul Karanicolas, John Somoza, Amber D. Wang, Clay C. C. Miao, Yinglong De Guzman, Roberto Oakley, Berl R. Neufeld, Kristi L. Xu, Liang Cancers (Basel) Article RNA-binding protein Musashi-1 (MSI1) is a key regulator of several stem cell populations. MSI1 is involved in tumor proliferation and maintenance, and it regulates target mRNAs at the translational level. The known mRNA targets of MSI1 include Numb, APC, and P21(WAF-1), key regulators of Notch/Wnt signaling and cell cycle progression, respectively. In this study, we aim to identify small molecule inhibitors of MSI1–mRNA interactions, which could block the growth of cancer cells with high levels of MSI1. Using a fluorescence polarization (FP) assay, we screened small molecules from several chemical libraries for those that disrupt the binding of MSI1 to its consensus RNA. One cluster of hit compounds is the derivatives of secondary metabolites from Aspergillus nidulans. One of the top hits, Aza-9, from this cluster was further validated by surface plasmon resonance and nuclear magnetic resonance spectroscopy, which demonstrated that Aza-9 binds directly to MSI1, and the binding is at the RNA binding pocket. We also show that Aza-9 binds to Musashi-2 (MSI2) as well. To test whether Aza-9 has anti-cancer potential, we used liposomes to facilitate Aza-9 cellular uptake. Aza-9-liposome inhibits proliferation, induces apoptosis and autophagy, and down-regulates Notch and Wnt signaling in colon cancer cell lines. In conclusion, we identified a series of potential lead compounds for inhibiting MSI1/2 function, while establishing a framework for identifying small molecule inhibitors of RNA binding proteins using FP-based screening methodology. MDPI 2020-08-08 /pmc/articles/PMC7463734/ /pubmed/32784494 http://dx.doi.org/10.3390/cancers12082221 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Lan, Lan Liu, Jiajun Xing, Minli Smith, Amber R. Wang, Jinan Wu, Xiaoqing Appelman, Carl Li, Ke Roy, Anuradha Gowthaman, Ragul Karanicolas, John Somoza, Amber D. Wang, Clay C. C. Miao, Yinglong De Guzman, Roberto Oakley, Berl R. Neufeld, Kristi L. Xu, Liang Identification and Validation of an Aspergillus nidulans Secondary Metabolite Derivative as an Inhibitor of the Musashi-RNA Interaction |
title | Identification and Validation of an Aspergillus nidulans Secondary Metabolite Derivative as an Inhibitor of the Musashi-RNA Interaction |
title_full | Identification and Validation of an Aspergillus nidulans Secondary Metabolite Derivative as an Inhibitor of the Musashi-RNA Interaction |
title_fullStr | Identification and Validation of an Aspergillus nidulans Secondary Metabolite Derivative as an Inhibitor of the Musashi-RNA Interaction |
title_full_unstemmed | Identification and Validation of an Aspergillus nidulans Secondary Metabolite Derivative as an Inhibitor of the Musashi-RNA Interaction |
title_short | Identification and Validation of an Aspergillus nidulans Secondary Metabolite Derivative as an Inhibitor of the Musashi-RNA Interaction |
title_sort | identification and validation of an aspergillus nidulans secondary metabolite derivative as an inhibitor of the musashi-rna interaction |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7463734/ https://www.ncbi.nlm.nih.gov/pubmed/32784494 http://dx.doi.org/10.3390/cancers12082221 |
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