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

Author:

Páscoa Inês1,Biltes Rita123ORCID,Sousa João123,Preto Marco Aurélio Correia1ORCID,Vasconcelos Vitor13ORCID,Castro Luís Filipe13ORCID,Ruivo Raquel1ORCID,Cunha Isabel1ORCID

Affiliation:

1. CIIMAR/CIMAR-Interdisciplinary Centre of Marine and Environmental Research, University of Porto, 4450-208 Matosinhos, Portugal

2. ICBAS-Instituto de Ciências Biomédicas Abel Salazar, University of Porto, 4050-313 Porto, Portugal

3. FCUP-Faculty of Sciences, Department of Biology, University of Porto, 4169-007 Porto, Portugal

Abstract

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.

Funder

FCT—Foundation for Science and Technology

CY-SENSORS

COMPETE 2020

Portugal 2020

European Union

ERDF

Publisher

MDPI AG

Subject

Electrical and Electronic Engineering,Biochemistry,Instrumentation,Atomic and Molecular Physics, and Optics,Analytical Chemistry

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