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A Multiplex Molecular Cell-Based Sensor to Detect Ligands of PPARs: An Optimized Tool for Drug Discovery in Cyanobacteria

Cyanobacteria produce a wealth of secondary metabolites. Since these organisms attach fatty acids into molecules in unprecedented ways, cyanobacteria can serve as a novel source for bioactive compounds acting as ligands for Peroxisome Proliferator-Activated Receptors (PPAR). PPARs (PPARα, PPARβ/δ an...

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Autores principales: Páscoa, Inês, Biltes, Rita, Sousa, João, Preto, Marco Aurélio Correia, Vasconcelos, Vitor, Castro, Luís Filipe, Ruivo, Raquel, Cunha, Isabel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9919141/
https://www.ncbi.nlm.nih.gov/pubmed/36772378
http://dx.doi.org/10.3390/s23031338
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author Páscoa, Inês
Biltes, Rita
Sousa, João
Preto, Marco Aurélio Correia
Vasconcelos, Vitor
Castro, Luís Filipe
Ruivo, Raquel
Cunha, Isabel
author_facet Páscoa, Inês
Biltes, Rita
Sousa, João
Preto, Marco Aurélio Correia
Vasconcelos, Vitor
Castro, Luís Filipe
Ruivo, Raquel
Cunha, Isabel
author_sort Páscoa, Inês
collection PubMed
description Cyanobacteria produce a wealth of secondary metabolites. Since these organisms attach fatty acids into molecules in unprecedented ways, cyanobacteria can serve as a novel source for bioactive compounds acting as ligands for Peroxisome Proliferator-Activated Receptors (PPAR). PPARs (PPARα, PPARβ/δ and PPARγ) are ligand-activated nuclear receptors, involved in the regulation of various metabolic and cellular processes, thus serving as potential drug targets for a variety of pathologies. Yet, given that PPARs’ agonists can have pan-, dual- or isoform-specific action, some controversy has been raised over currently approved drugs and their side effects, highlighting the need for novel molecules. Here, we expand and validate a cell-based PPAR transactivation activity biosensor, and test it in a screening campaign to guide drug discovery. Biosensor upgrades included the use of different reporter genes to increase signal intensity and stability, a different promoter to modulate reporter gene expression, and multiplexing to improve efficiency. Sensor’s limit of detection (LOD) ranged from 0.36–0.89 nM in uniplex and 0.89–1.35 nM in multiplex mode. In triplex mode, the sensor’s feature screening, a total of 848 fractions of 96 cyanobacteria extracts were screened. Hits were confirmed in multiplex mode and in uniplex mode, yielding one strain detected to have action on PPARα and three strains to have dual action on PPARα and -β.
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spelling pubmed-99191412023-02-12 A Multiplex Molecular Cell-Based Sensor to Detect Ligands of PPARs: An Optimized Tool for Drug Discovery in Cyanobacteria Páscoa, Inês Biltes, Rita Sousa, João Preto, Marco Aurélio Correia Vasconcelos, Vitor Castro, Luís Filipe Ruivo, Raquel Cunha, Isabel Sensors (Basel) Article Cyanobacteria produce a wealth of secondary metabolites. Since these organisms attach fatty acids into molecules in unprecedented ways, cyanobacteria can serve as a novel source for bioactive compounds acting as ligands for Peroxisome Proliferator-Activated Receptors (PPAR). PPARs (PPARα, PPARβ/δ and PPARγ) are ligand-activated nuclear receptors, involved in the regulation of various metabolic and cellular processes, thus serving as potential drug targets for a variety of pathologies. Yet, given that PPARs’ agonists can have pan-, dual- or isoform-specific action, some controversy has been raised over currently approved drugs and their side effects, highlighting the need for novel molecules. Here, we expand and validate a cell-based PPAR transactivation activity biosensor, and test it in a screening campaign to guide drug discovery. Biosensor upgrades included the use of different reporter genes to increase signal intensity and stability, a different promoter to modulate reporter gene expression, and multiplexing to improve efficiency. Sensor’s limit of detection (LOD) ranged from 0.36–0.89 nM in uniplex and 0.89–1.35 nM in multiplex mode. In triplex mode, the sensor’s feature screening, a total of 848 fractions of 96 cyanobacteria extracts were screened. Hits were confirmed in multiplex mode and in uniplex mode, yielding one strain detected to have action on PPARα and three strains to have dual action on PPARα and -β. MDPI 2023-01-25 /pmc/articles/PMC9919141/ /pubmed/36772378 http://dx.doi.org/10.3390/s23031338 Text en © 2023 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
Páscoa, Inês
Biltes, Rita
Sousa, João
Preto, Marco Aurélio Correia
Vasconcelos, Vitor
Castro, Luís Filipe
Ruivo, Raquel
Cunha, Isabel
A Multiplex Molecular Cell-Based Sensor to Detect Ligands of PPARs: An Optimized Tool for Drug Discovery in Cyanobacteria
title A Multiplex Molecular Cell-Based Sensor to Detect Ligands of PPARs: An Optimized Tool for Drug Discovery in Cyanobacteria
title_full A Multiplex Molecular Cell-Based Sensor to Detect Ligands of PPARs: An Optimized Tool for Drug Discovery in Cyanobacteria
title_fullStr A Multiplex Molecular Cell-Based Sensor to Detect Ligands of PPARs: An Optimized Tool for Drug Discovery in Cyanobacteria
title_full_unstemmed A Multiplex Molecular Cell-Based Sensor to Detect Ligands of PPARs: An Optimized Tool for Drug Discovery in Cyanobacteria
title_short A Multiplex Molecular Cell-Based Sensor to Detect Ligands of PPARs: An Optimized Tool for Drug Discovery in Cyanobacteria
title_sort multiplex molecular cell-based sensor to detect ligands of ppars: an optimized tool for drug discovery in cyanobacteria
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9919141/
https://www.ncbi.nlm.nih.gov/pubmed/36772378
http://dx.doi.org/10.3390/s23031338
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