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Droplet Digital PCR Is a Novel Screening Method Identifying Potential Cardiac G-Protein-Coupled Receptors as Candidate Pharmacological Targets in a Rat Model of Pressure-Overload-Induced Cardiac Dysfunction
The identification of novel drug targets is needed to improve the outcomes of heart failure (HF). G-protein-coupled receptors (GPCRs) represent the largest family of targets for already approved drugs, thus providing an opportunity for drug repurposing. Here, we aimed (i) to investigate the differen...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10531061/ https://www.ncbi.nlm.nih.gov/pubmed/37762130 http://dx.doi.org/10.3390/ijms241813826 |
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author | Sayour, Nabil V. Tóth, Viktória É. Nagy, Regina N. Vörös, Imre Gergely, Tamás G. Onódi, Zsófia Nagy, Noémi Bödör, Csaba Váradi, Barnabás Ruppert, Mihály Radovits, Tamás Bleckwedel, Federico Zelarayán, Laura C. Pacher, Pal Ágg, Bence Görbe, Anikó Ferdinandy, Péter Varga, Zoltán V. |
author_facet | Sayour, Nabil V. Tóth, Viktória É. Nagy, Regina N. Vörös, Imre Gergely, Tamás G. Onódi, Zsófia Nagy, Noémi Bödör, Csaba Váradi, Barnabás Ruppert, Mihály Radovits, Tamás Bleckwedel, Federico Zelarayán, Laura C. Pacher, Pal Ágg, Bence Görbe, Anikó Ferdinandy, Péter Varga, Zoltán V. |
author_sort | Sayour, Nabil V. |
collection | PubMed |
description | The identification of novel drug targets is needed to improve the outcomes of heart failure (HF). G-protein-coupled receptors (GPCRs) represent the largest family of targets for already approved drugs, thus providing an opportunity for drug repurposing. Here, we aimed (i) to investigate the differential expressions of 288 cardiac GPCRs via droplet digital PCR (ddPCR) and bulk RNA sequencing (RNAseq) in a rat model of left ventricular pressure-overload; (ii) to compare RNAseq findings with those of ddPCR; and (iii) to screen and test for novel, translatable GPCR drug targets in HF. Male Wistar rats subjected to transverse aortic constriction (TAC, n = 5) showed significant systolic dysfunction vs. sham operated animals (SHAM, n = 5) via echocardiography. In TAC vs. SHAM hearts, RNAseq identified 69, and ddPCR identified 27 significantly differentially expressed GPCR mRNAs, 8 of which were identified using both methods, thus showing a correlation between the two methods. Of these, Prostaglandin-F2α-receptor (Ptgfr) was further investigated and localized on cardiomyocytes and fibroblasts in murine hearts via RNA-Scope. Antagonizing Ptgfr via AL-8810 reverted angiotensin-II-induced cardiomyocyte hypertrophy in vitro. In conclusion, using ddPCR as a novel screening method, we were able to identify GPCR targets in HF. We also show that the antagonism of Ptgfr could be a novel target in HF by alleviating cardiomyocyte hypertrophy. |
format | Online Article Text |
id | pubmed-10531061 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-105310612023-09-28 Droplet Digital PCR Is a Novel Screening Method Identifying Potential Cardiac G-Protein-Coupled Receptors as Candidate Pharmacological Targets in a Rat Model of Pressure-Overload-Induced Cardiac Dysfunction Sayour, Nabil V. Tóth, Viktória É. Nagy, Regina N. Vörös, Imre Gergely, Tamás G. Onódi, Zsófia Nagy, Noémi Bödör, Csaba Váradi, Barnabás Ruppert, Mihály Radovits, Tamás Bleckwedel, Federico Zelarayán, Laura C. Pacher, Pal Ágg, Bence Görbe, Anikó Ferdinandy, Péter Varga, Zoltán V. Int J Mol Sci Article The identification of novel drug targets is needed to improve the outcomes of heart failure (HF). G-protein-coupled receptors (GPCRs) represent the largest family of targets for already approved drugs, thus providing an opportunity for drug repurposing. Here, we aimed (i) to investigate the differential expressions of 288 cardiac GPCRs via droplet digital PCR (ddPCR) and bulk RNA sequencing (RNAseq) in a rat model of left ventricular pressure-overload; (ii) to compare RNAseq findings with those of ddPCR; and (iii) to screen and test for novel, translatable GPCR drug targets in HF. Male Wistar rats subjected to transverse aortic constriction (TAC, n = 5) showed significant systolic dysfunction vs. sham operated animals (SHAM, n = 5) via echocardiography. In TAC vs. SHAM hearts, RNAseq identified 69, and ddPCR identified 27 significantly differentially expressed GPCR mRNAs, 8 of which were identified using both methods, thus showing a correlation between the two methods. Of these, Prostaglandin-F2α-receptor (Ptgfr) was further investigated and localized on cardiomyocytes and fibroblasts in murine hearts via RNA-Scope. Antagonizing Ptgfr via AL-8810 reverted angiotensin-II-induced cardiomyocyte hypertrophy in vitro. In conclusion, using ddPCR as a novel screening method, we were able to identify GPCR targets in HF. We also show that the antagonism of Ptgfr could be a novel target in HF by alleviating cardiomyocyte hypertrophy. MDPI 2023-09-07 /pmc/articles/PMC10531061/ /pubmed/37762130 http://dx.doi.org/10.3390/ijms241813826 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 Sayour, Nabil V. Tóth, Viktória É. Nagy, Regina N. Vörös, Imre Gergely, Tamás G. Onódi, Zsófia Nagy, Noémi Bödör, Csaba Váradi, Barnabás Ruppert, Mihály Radovits, Tamás Bleckwedel, Federico Zelarayán, Laura C. Pacher, Pal Ágg, Bence Görbe, Anikó Ferdinandy, Péter Varga, Zoltán V. Droplet Digital PCR Is a Novel Screening Method Identifying Potential Cardiac G-Protein-Coupled Receptors as Candidate Pharmacological Targets in a Rat Model of Pressure-Overload-Induced Cardiac Dysfunction |
title | Droplet Digital PCR Is a Novel Screening Method Identifying Potential Cardiac G-Protein-Coupled Receptors as Candidate Pharmacological Targets in a Rat Model of Pressure-Overload-Induced Cardiac Dysfunction |
title_full | Droplet Digital PCR Is a Novel Screening Method Identifying Potential Cardiac G-Protein-Coupled Receptors as Candidate Pharmacological Targets in a Rat Model of Pressure-Overload-Induced Cardiac Dysfunction |
title_fullStr | Droplet Digital PCR Is a Novel Screening Method Identifying Potential Cardiac G-Protein-Coupled Receptors as Candidate Pharmacological Targets in a Rat Model of Pressure-Overload-Induced Cardiac Dysfunction |
title_full_unstemmed | Droplet Digital PCR Is a Novel Screening Method Identifying Potential Cardiac G-Protein-Coupled Receptors as Candidate Pharmacological Targets in a Rat Model of Pressure-Overload-Induced Cardiac Dysfunction |
title_short | Droplet Digital PCR Is a Novel Screening Method Identifying Potential Cardiac G-Protein-Coupled Receptors as Candidate Pharmacological Targets in a Rat Model of Pressure-Overload-Induced Cardiac Dysfunction |
title_sort | droplet digital pcr is a novel screening method identifying potential cardiac g-protein-coupled receptors as candidate pharmacological targets in a rat model of pressure-overload-induced cardiac dysfunction |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10531061/ https://www.ncbi.nlm.nih.gov/pubmed/37762130 http://dx.doi.org/10.3390/ijms241813826 |
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