Activation of phosphodiesterase type 2 to treat heart failure Marta Lindner To cite this version: Marta Lindner. Activation of phosphodiesterase type 2 to treat heart failure. Tissues and Organs [q-bio.TO]. Université Paris Saclay (COmUE), 2016. English. NNT : 2016SACLS336. tel-01877874 HAL Id: tel-01877874 https://tel.archives-ouvertes.fr/tel-01877874 Submitted on 20 Sep 2018 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. NNT : 2016SACL336 THESE DE DOCTORAT DE L’UNIVERSITE PARIS-SACLAY PREPAREE A UNIVERSITE PARIS SUD ECOLE DOCTORALE N° 569 ITFA Innovation thérapeutique: du fondamental à l’appliqué Spécialité de doctorat: Physiologie, physiopathologie Par Marta Lindner Activation de la phosphodiestérase de type 2 pour traiter l'insuffisance cardiaque Thèse présentée et soutenue à Châtenay-Malabry, le 12 Octobre 2016 Composition du Jury : M. Christian POÜS Professeur, UPSud Président Mme Claire LUGNIER DR Emérite CNRS Rapporteur Mme Catherine PAVOINE CR INSERM Rapporteur Mme Liliana CASTRO Maître de Conférence, UPMC Examinateur M. Ali EL-ARMOUCHE Professeur, Univ. Dresden Examinateur M. Grégoire VANDECASTEELE DR INSERM Examinateur M. Rodolphe FISCHMEISTER DR INSERM Directeur de thèse Table of contents Sommaire Introduction ______________________________________________________________ 13 I. The heart _______________________________________________________________ 14 I.1 Anatomy of the heart ________________________________________________________ 14 I.2 Cardiac excitation-contraction coupling __________________________________________ 15 I.2.1 Structures involved in cardiac ECC _____________________________________________________ 15 I.2.1.1 Sarcomere and T-tubules _______________________________________________________ 15 I.2.1.2 Sarcoplasmic reticulum ________________________________________________________ 17 I.2.1.3 Myofilaments ________________________________________________________________ 17 I.2.1.4 Mitochondria ________________________________________________________________ 18 I.2.2 Excitation _________________________________________________________________________ 19 I.2.3 Contraction _______________________________________________________________________ 21 I.2.4 Relaxation ________________________________________________________________________ 22 I.3 Regulation of ECC ____________________________________________________________ 22 I.3.1 Frank-Starling law __________________________________________________________________ 22 I.3.2 Neurohormonal regulation of heart function ____________________________________________ 23 II. Cyclic AMP signaling in the heart ___________________________________________ 24 II.1 G-protein coupled receptors (GPCRs) ___________________________________________ 24 II.2 β-adrenergic receptors _______________________________________________________ 25 II.2.1 β1-adrenergic receptor ______________________________________________________________ 27 II.2.2 β2-adrenergic receptors _____________________________________________________________ 28 II.2.3 β3-adrenergic receptors _____________________________________________________________ 29 II.2.4 Desensitization of β-adrenergic receptors ______________________________________________ 29 II.3 Adenylyl cyclases ___________________________________________________________ 31 II.4 Effectors of cAMP ___________________________________________________________ 34 II.4.1 Protein kinase A ___________________________________________________________________ 34 II.4.1.1 Structure and regulation _______________________________________________________ 34 II.4.1.2 Targets of PKA in the heart _____________________________________________________ 35 L-type Ca2+ channels (LTTCs) ____________________________________________________ 35 Ryanodine receptor ___________________________________________________________ 37 Phospholamban ______________________________________________________________ 39 Contractile proteins___________________________________________________________ 39 II.4.1.3 Subcellular targeting of PKA ____________________________________________________ 40 II.4.2 Other effectors of cAMP ____________________________________________________________ 40 II.4.2.1 Exchange factor Epac _________________________________________________________ 40 II.4.2.2 CNG and HCN channels ________________________________________________________ 42 II.4.2.3 Popeye domain containing protein family _________________________________________ 43 II.5 cAMP degradation __________________________________________________________ 43 II.5.1 Structure, localization and regulation of phosphodiesterases _______________________________ 45 II.5.1.1 Phosphodiesterase type 1 ______________________________________________________ 45 II.5.1.2 Phosphodiesterase type 2 ______________________________________________________ 47 II.5.1.3 Phosphodiesterase type 3 ______________________________________________________ 48 1 II.5.1.4 Phosphodiesterase type 4 ______________________________________________________ 49 II.5.2 Pharmacological inhibition of phosphodiesterases _______________________________________ 50 II.5.3 cAMP compartmentation ____________________________________________________________ 51 III. Cyclic GMP signaling in the heart ___________________________________________ 54 III.1 Cyclic GMP production _______________________________________________________ 54 III.1.1 Soluble guanylyl cyclase ____________________________________________________________ 55 III.1.2 Particulate guanylyl cyclase _________________________________________________________ 56 III.2 cGMP targets in cardiac myocytes _____________________________________________ 57 III.2.1 Protein kinase G __________________________________________________________________ 57 III.2.1.1 LTCCs ______________________________________________________________________ 59 III.2.1.2 Troponin I __________________________________________________________________ 59 III.2.1.3 Phospholamban _____________________________________________________________ 60 III.2.2 Phosphodiesterase ________________________________________________________________ 60 III.3 cGMP signaling in cardiac hypertrophy__________________________________________ 62 IV. cAMP and cGMP signaling crosstalk ________________________________________ 63 V. Heart failure ____________________________________________________________ 66 V.1 Pathophysiology of heart failure _______________________________________________ 66 V.2 Treatment of heart failure ____________________________________________________ 67 V.2.1 β-blockers ________________________________________________________________________ 67 V.2.2 Inhibitors of phosphodiesterases _____________________________________________________ 68 V.2.3 Vasodilators ______________________________________________________________________ 69 V.2.4 Ca2+ channel blockers _______________________________________________________________ 69 V.2.5 ACE inhibitors and angiotensin receptor blockers ________________________________________ 70 V.2.6 LCZ 696 __________________________________________________________________________ 70 VI. Phosphodiesterase type 2 _________________________________________________ 71 VI.1 Structure, function and subcellular localization of PDE2 ____________________________ 71 VI.2 Regulation of expression and activity of PDE2 ____________________________________ 73 VI.3 PDE2 as potential target in heart failure ________________________________________ 74 Objectives ________________________________________________________________ 76 Materials and methods _____________________________________________________ 79 I. Animal model ____________________________________________________________ 80 I.1 Transgenic mice with PDE2 overexpression _______________________________________ 80 I.2 Experimental model of hypertrophy _____________________________________________ 80 II. Exercise capacity test _____________________________________________________ 81 III. Langendorff perfused heart _______________________________________________ 82 III.1 Principle __________________________________________________________________ 82 III.2 Experimental set-up _________________________________________________________ 82 III.3 Experimental protocol _______________________________________________________ 83 IV. Isolation of adult mouse ventricular cardiomyocytes ___________________________ 84 2 V. Sarcomere shortening and Ca2+ transient measurements ________________________ 85 V.1 Principle __________________________________________________________________ 85 V.2 Fluorescent Ca2+ indicator Fura-2-AM ___________________________________________ 85 V.3 Experimental set-up _________________________________________________________ 86 V.4 Experimental protocols ______________________________________________________ 87 V.5 Data processing _____________________________________________________________ 88 V.6 Data analysis _______________________________________________________________ 88 VI. Fluorescence resonance energy transfer (FRET) imaging ________________________ 89