title: Paracrine nitric oxide induces expression of cardiac sarcomeric proteins in adult progenitor cells through soluble guanylyl cyclase/cyclic-guanosine monophosphate and Wnt/β-catenin inhibition.
abstract: Cardiac progenitor cells (CPC) from adult hearts can differentiate to several cell types composing the myocardium but the underlying molecular pathways are poorly characterized. We examined the role of paracrine nitric oxide (NO) in the specification of CPC to the cardiac lineage, particularly through its inhibition of the canonical Wnt/β-catenin pathway, a critical step preceding cardiac differentiation.
  
  
  
  Sca1 + CPC from adult mouse hearts were isolated by magnetic-activated cell sorting and clonally expanded. Pharmacologic NO donors increased their expression of cardiac myocyte-specific sarcomeric proteins in a concentration and time-dependent manner. The optimal time window for NO efficacy coincided with up-regulation of CPC expression of Gucy1a3 (coding the alpha1 subunit of guanylyl cyclase). The effect of paracrine NO was reproduced in vitro upon co-culture of CPC with cardiac myocytes expressing a transgenic NOS3 (endothelial nitric oxide synthase) and in vivo upon injection of CPC in infarcted hearts from cardiac-specific NOS3 transgenic mice. In mono- and co-cultures, this effect was abrogated upon inhibition of soluble guanylyl cyclase or nitric oxide synthase, and was lost in CPC genetically deficient in Gucy1a3. Mechanistically, NO inhibits the constitutive activity of the canonical Wnt/β-catenin in CPC and in cell reporter assays in a guanylyl cyclase-dependent fashion. This was paralleled with decreased expression of β-catenin and down-regulation of Wnt target genes in CPC and abrogated in CPC with a stabilized, non-inhibitable β-catenin.
  
  
  
  Exogenous or paracrine sources of NO promote the specification towards the myocyte lineage and expression of cardiac sarcomeric proteins of adult CPC. This is contingent upon the expression and activity of the alpha1 subunit of guanylyl cyclase in CPC that is necessary for NO-mediated inhibition of the canonical Wnt/β-catenin pathway.
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Aurelia
authors: A
authors: De Pauw
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Paul
authors: P
authors: Massion
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Belaid
authors: B
authors: Sekkali
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Emilie
authors: E
authors: Andre
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Caroline
authors: C
authors: Dubroca
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Jana
authors: J
authors: Kmecova
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Caroline
authors: C
authors: Bouzin
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Ann
authors: A
authors: Friart
authors: Department of Human Genetics, Cliniques Saint-Luc, Université Catholique de Louvain, 10 avenue Hippocrate, 1200 Brussels, Belgium.
authors: Catherine
authors: C
authors: Sibille
authors: Pole of Endocrinology, Diabetes and Nutrition (EDIN), Institut de Recherche Expérimentale et Clinique (IREC), Université Catholique de Louvain, B1.55.06, 55 avenue Hippocrate, 1200 Brussels, Belgium.
authors: Patrick
authors: P
authors: Gilon
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Delphine
authors: D
authors: De Mulder
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Hrag
authors: H
authors: Esfahani
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Adrien
authors: A
authors: Strapart
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Ruben
authors: R
authors: Martherus
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Valéry
authors: V
authors: Payen
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium.
authors: Pierre
authors: P
authors: Sonveaux
authors: Department of Biomedical Molecular Biology, Universiteit Gent, Technologiepark 927, 9052 Gent, Belgium.
authors: Peter
authors: P
authors: Brouckaert
authors: Department of Cardiology, Katholieke Universiteit Leuven, Herestraat 49, 3000 Leuven, Belgium.
authors: Stefan
authors: S
authors: Janssens
authors: Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Departement de Medecine Interne et Cliniques Saint-Luc, Université Catholique de Louvain, B1.53.09, 52 Ave. Mounier, 1200 Brussels, Belgium jl.balligand@uclouvain.be.
authors: Jean-Luc
authors: JL
authors: Balligand
mesh_terms: D053687:Adult Stem Cells / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D000950:Antigens, Ly / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage; D002478:Cells, Cultured; D018920:Coculture Techniques; D006152:Cyclic GMP / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D018189:Immunomagnetic Separation; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000201:enzymology*; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D019899:Paracrine Communication / Q000187:drug effects; D012518:Sarcomeres / Q000187:drug effects / Q000201:enzymology*; D015398:Signal Transduction; D000071756:Soluble Guanylyl Cyclase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D060449:Wnt Signaling Pathway / Q000187:drug effects; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism*
chemical_list: D000950:Antigens, Ly; C495267:CTNNB1 protein, mouse; C000605668:Gucy1a3 protein, mouse; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D020030:Nitric Oxide Donors; D051176:beta Catenin; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D000071756:Soluble Guanylyl Cyclase; D006152:Cyclic GMP
keywords: Cardiac progenitor cells; Cell–cell communication; Guanylyl cyclase; Nitric oxide; Wnt pathway