The identification of adenylyl cyclase modulators as potential receptors for 6-nitrodopamine in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and their relevance in heart inotropism

6-Nitrodopamine (6-ND) has potent positive chronotropic and inotropic effects. At a very low dose, i.e., 10 fM, it causes potentiation of the positive chronotropic effects induced by catecholamines in the rat atria, indicating a distinct mechanism of action. Cyclase-associated proteins (CAP-1 and CA...

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Main Authors: Irene Cipollone, Vittoria Monaco, José Britto-Júnior, Antonio Tiago Lima, Edson Antunes, Andre Sampaio Pupo, Ilaria Iacobucci, Flora Cozzolino, Maria Monti, Silvia Parisi, Giuseppina Divisato, Emanuela Cascone, Alfonso De Simone, Angela Corvino, Ferdinando Fiorino, Francesco Frecentese, Vincenzo Santagada, Beatrice Severino, Rosa Sparaco, Pierfrancesco Cinque, Stefania Vertuccio, Giuseppe Caliendo, Gilberto De Nucci
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Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Pharmacology
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Online Access:https://www.frontiersin.org/articles/10.3389/fphar.2025.1597035/full
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author Irene Cipollone
Irene Cipollone
Vittoria Monaco
Vittoria Monaco
José Britto-Júnior
Antonio Tiago Lima
Edson Antunes
Andre Sampaio Pupo
Ilaria Iacobucci
Ilaria Iacobucci
Flora Cozzolino
Flora Cozzolino
Maria Monti
Maria Monti
Silvia Parisi
Giuseppina Divisato
Emanuela Cascone
Alfonso De Simone
Angela Corvino
Ferdinando Fiorino
Francesco Frecentese
Vincenzo Santagada
Beatrice Severino
Rosa Sparaco
Pierfrancesco Cinque
Stefania Vertuccio
Giuseppe Caliendo
Gilberto De Nucci
Gilberto De Nucci
Gilberto De Nucci
author_facet Irene Cipollone
Irene Cipollone
Vittoria Monaco
Vittoria Monaco
José Britto-Júnior
Antonio Tiago Lima
Edson Antunes
Andre Sampaio Pupo
Ilaria Iacobucci
Ilaria Iacobucci
Flora Cozzolino
Flora Cozzolino
Maria Monti
Maria Monti
Silvia Parisi
Giuseppina Divisato
Emanuela Cascone
Alfonso De Simone
Angela Corvino
Ferdinando Fiorino
Francesco Frecentese
Vincenzo Santagada
Beatrice Severino
Rosa Sparaco
Pierfrancesco Cinque
Stefania Vertuccio
Giuseppe Caliendo
Gilberto De Nucci
Gilberto De Nucci
Gilberto De Nucci
author_sort Irene Cipollone
collection DOAJ
description 6-Nitrodopamine (6-ND) has potent positive chronotropic and inotropic effects. At a very low dose, i.e., 10 fM, it causes potentiation of the positive chronotropic effects induced by catecholamines in the rat atria, indicating a distinct mechanism of action. Cyclase-associated proteins (CAP-1 and CAP-2) are potential receptors for 6-ND in human cardiomyocytes. Since cyclic 3′,5′-cyclic adenosine monophosphate (cAMP) plays a fundamental role in the positive inotropic effects of classical catecholamines, it was further investigated whether 6-ND potentiates the positive inotropic effects induced by classical catecholamines in the rat isolated perfused heart. Human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes were harvested and lysed, and following appropriate separation procedures, membrane proteins were incubated with 6-ND-derivatized agarose, centrifuged, and the proteins retained in the agarose eluted with 6-ND (1 mM). The proteins isolated from the chemical pulldown assay were fractionated by SDS-PAGE, the bands were cut and hydrolyzed in situ with trypsin, and then separated and sequenced. A total of 817 proteins were generated, and following screening using UniProt “Retrieve/ID Mapping” function and Gene Ontology cellular component category, 124 proteins were identified as membrane proteins. These experiments identified three proteins that modulate adenylyl cyclase (AC) activity (CAP-1, CAP-2, and STIM1), which are compatible with the pharmacological findings reported for 6-ND in the rat heart. As expected, 6-ND strongly potentiates the inotropic effect induced by noradrenaline in Langendorff’s preparation. In conclusion, 6-ND-induced potentiation of catecholamine-induced chronotropic and inotropic effects is due to the modulation of adenylyl cyclase activity, probably via direct interactions with CAP-1 and CAP-2.
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spelling doaj-art-64cddf11cb2c49138d0cd80c4a0c26db2025-08-20T04:02:27ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-08-011610.3389/fphar.2025.15970351597035The identification of adenylyl cyclase modulators as potential receptors for 6-nitrodopamine in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and their relevance in heart inotropismIrene Cipollone0Irene Cipollone1Vittoria Monaco2Vittoria Monaco3José Britto-Júnior4Antonio Tiago Lima5Edson Antunes6Andre Sampaio Pupo7Ilaria Iacobucci8Ilaria Iacobucci9Flora Cozzolino10Flora Cozzolino11Maria Monti12Maria Monti13Silvia Parisi14Giuseppina Divisato15Emanuela Cascone16Alfonso De Simone17Angela Corvino18Ferdinando Fiorino19Francesco Frecentese20Vincenzo Santagada21Beatrice Severino22Rosa Sparaco23Pierfrancesco Cinque24Stefania Vertuccio25Giuseppe Caliendo26Gilberto De Nucci27Gilberto De Nucci28Gilberto De Nucci29Department of Chemical Sciences, University of Naples Federico II, Naples, ItalyCEINGE Biotecnologie Avanzate “Franco Salvatore”, Naples, ItalyDepartment of Chemical Sciences, University of Naples Federico II, Naples, ItalyCEINGE Biotecnologie Avanzate “Franco Salvatore”, Naples, ItalyDepartment of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), São Paulo, BrazilDepartment of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), São Paulo, BrazilDepartment of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), São Paulo, BrazilDepartment of Biophysics and Pharmacology, Institute of Biosciences of Botucatu, São Paulo State University (UNESP), Botucatu, BrazilDepartment of Chemical Sciences, University of Naples Federico II, Naples, ItalyCEINGE Biotecnologie Avanzate “Franco Salvatore”, Naples, ItalyDepartment of Chemical Sciences, University of Naples Federico II, Naples, ItalyCEINGE Biotecnologie Avanzate “Franco Salvatore”, Naples, ItalyDepartment of Chemical Sciences, University of Naples Federico II, Naples, ItalyCEINGE Biotecnologie Avanzate “Franco Salvatore”, Naples, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Biophysics and Pharmacology, Institute of Biosciences of Botucatu, São Paulo State University (UNESP), Botucatu, BrazilDepartment of Pharmacy, School of Medicine, University of Naples Federico II, Naples, ItalyDepartment of Pharmacology, Institute of Biomedical Sciences, University of São Paulo (ICB-USP), São Paulo, Brazil6-Nitrodopamine (6-ND) has potent positive chronotropic and inotropic effects. At a very low dose, i.e., 10 fM, it causes potentiation of the positive chronotropic effects induced by catecholamines in the rat atria, indicating a distinct mechanism of action. Cyclase-associated proteins (CAP-1 and CAP-2) are potential receptors for 6-ND in human cardiomyocytes. Since cyclic 3′,5′-cyclic adenosine monophosphate (cAMP) plays a fundamental role in the positive inotropic effects of classical catecholamines, it was further investigated whether 6-ND potentiates the positive inotropic effects induced by classical catecholamines in the rat isolated perfused heart. Human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes were harvested and lysed, and following appropriate separation procedures, membrane proteins were incubated with 6-ND-derivatized agarose, centrifuged, and the proteins retained in the agarose eluted with 6-ND (1 mM). The proteins isolated from the chemical pulldown assay were fractionated by SDS-PAGE, the bands were cut and hydrolyzed in situ with trypsin, and then separated and sequenced. A total of 817 proteins were generated, and following screening using UniProt “Retrieve/ID Mapping” function and Gene Ontology cellular component category, 124 proteins were identified as membrane proteins. These experiments identified three proteins that modulate adenylyl cyclase (AC) activity (CAP-1, CAP-2, and STIM1), which are compatible with the pharmacological findings reported for 6-ND in the rat heart. As expected, 6-ND strongly potentiates the inotropic effect induced by noradrenaline in Langendorff’s preparation. In conclusion, 6-ND-induced potentiation of catecholamine-induced chronotropic and inotropic effects is due to the modulation of adenylyl cyclase activity, probably via direct interactions with CAP-1 and CAP-2.https://www.frontiersin.org/articles/10.3389/fphar.2025.1597035/fulldopamineadrenalinenitro-catecholaminestromal interaction molecule 1cyclase-associated proteins
spellingShingle Irene Cipollone
Irene Cipollone
Vittoria Monaco
Vittoria Monaco
José Britto-Júnior
Antonio Tiago Lima
Edson Antunes
Andre Sampaio Pupo
Ilaria Iacobucci
Ilaria Iacobucci
Flora Cozzolino
Flora Cozzolino
Maria Monti
Maria Monti
Silvia Parisi
Giuseppina Divisato
Emanuela Cascone
Alfonso De Simone
Angela Corvino
Ferdinando Fiorino
Francesco Frecentese
Vincenzo Santagada
Beatrice Severino
Rosa Sparaco
Pierfrancesco Cinque
Stefania Vertuccio
Giuseppe Caliendo
Gilberto De Nucci
Gilberto De Nucci
Gilberto De Nucci
The identification of adenylyl cyclase modulators as potential receptors for 6-nitrodopamine in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and their relevance in heart inotropism
Frontiers in Pharmacology
dopamine
adrenaline
nitro-catecholamine
stromal interaction molecule 1
cyclase-associated proteins
title The identification of adenylyl cyclase modulators as potential receptors for 6-nitrodopamine in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and their relevance in heart inotropism
title_full The identification of adenylyl cyclase modulators as potential receptors for 6-nitrodopamine in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and their relevance in heart inotropism
title_fullStr The identification of adenylyl cyclase modulators as potential receptors for 6-nitrodopamine in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and their relevance in heart inotropism
title_full_unstemmed The identification of adenylyl cyclase modulators as potential receptors for 6-nitrodopamine in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and their relevance in heart inotropism
title_short The identification of adenylyl cyclase modulators as potential receptors for 6-nitrodopamine in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and their relevance in heart inotropism
title_sort identification of adenylyl cyclase modulators as potential receptors for 6 nitrodopamine in human induced pluripotent stem cell hipsc derived cardiomyocytes and their relevance in heart inotropism
topic dopamine
adrenaline
nitro-catecholamine
stromal interaction molecule 1
cyclase-associated proteins
url https://www.frontiersin.org/articles/10.3389/fphar.2025.1597035/full
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