title: Low doses of arginine butyrate derivatives improve dystrophic phenotype and restore membrane integrity in DMD models.
abstract: A new approach to treating Duchenne muscular dystrophy was investigated by using the ester or amide covalent association of arginine [nitric oxide (NO) pathway] and butyrate [histone deacetylase (HDAC) inhibition] in mdx mice and patient myotubes. Two prodrugs were synthesized, and the beneficial effects on dystrophic phenotype were studied. Nerve excitability abnormalities detected in saline-treated mice were almost totally rescued in animals treated at low doses (50-100 mg/kg/d). Force and fatigue resistance were improved ≈60% and 3.5-fold, respectively, and the percentage of necrosis in heart sections was reduced ≈90% in the treated mice. A decrease of >50% in serum creatine kinase indicated an overall improvement in the muscles. Restoration of membrane integrity was studied directly by measuring the reduction (≈74%) of Evans blue incorporation in the limb muscles of the treated animals, the increase in utrophin level, and the normalization of lipid composition of the heart. In cultures of human myotubes (primary cells and cell line), both prodrugs and HDAC inhibitors increased by 2- to 4-fold the utrophin level, which was correctly localized at the membrane. β-Dystroglycan and embryonic myosin protein levels were also increased. Finally, a 50% reduction in the number of spontaneous Ca(2+) spikes was observed after treatment with NO synthase substrate and HDAC inhibitors. Overall, the beneficial effects were obtained with doses 10 (in vivo) and 5 (in vitro) times lower than those of the salt formulation. Altogether, these data constitute proof of principle of the beneficial effects of low doses of arginine butyrate derivatives on muscular dystrophy, enhancing the NO pathway and inhibiting HDAC.
authors.affiliation: Neurobiologie and Développement, Institut de Neurobiologie Alfred Fessard-Fédération pour la Recherche sur le Cerveau (FRC) 2118, Unité Propres de Recherche (UPR) 3294, and.
authors.forename: Sara
authors.initials: S
authors.lastname: Vianello
authors.affiliation: Neurobiologie and Développement, Institut de Neurobiologie Alfred Fessard-Fédération pour la Recherche sur le Cerveau (FRC) 2118, Unité Propres de Recherche (UPR) 3294, and.
authors.forename: Francesca
authors.initials: F
authors.lastname: Consolaro
authors.affiliation: Institut de Chimie des Substances Naturelles, Centre de Recherche de Gif, UPR 2301, Centre National de la Recherche Scientifique (CNRS), Gif sur Yvette, France; and.
authors.forename: Claudia
authors.initials: C
authors.lastname: Bich
authors.affiliation: Centre de Neurosciences Paris-Sud, Université Paris Sud, Unité Mixte de Recherche (UMR) 8195, CNRS, Orsay, France.
authors.forename: José-Manuel
authors.initials: JM
authors.lastname: Cancela
authors.affiliation: Neurobiologie and Développement, Institut de Neurobiologie Alfred Fessard-Fédération pour la Recherche sur le Cerveau (FRC) 2118, Unité Propres de Recherche (UPR) 3294, and.
authors.forename: Morgane
authors.initials: M
authors.lastname: Roulot
authors.affiliation: Centre de Neurosciences Paris-Sud, Université Paris Sud, Unité Mixte de Recherche (UMR) 8195, CNRS, Orsay, France.
authors.forename: Erwan
authors.initials: E
authors.lastname: Lanchec
authors.affiliation: Institut de Chimie des Substances Naturelles, Centre de Recherche de Gif, UPR 2301, Centre National de la Recherche Scientifique (CNRS), Gif sur Yvette, France; and.
authors.forename: David
authors.initials: D
authors.lastname: Touboul
authors.affiliation: Institut de Chimie des Substances Naturelles, Centre de Recherche de Gif, UPR 2301, Centre National de la Recherche Scientifique (CNRS), Gif sur Yvette, France; and.
authors.forename: Alain
authors.initials: A
authors.lastname: Brunelle
authors.affiliation: Neurobiologie and Développement, Institut de Neurobiologie Alfred Fessard-Fédération pour la Recherche sur le Cerveau (FRC) 2118, Unité Propres de Recherche (UPR) 3294, and.
authors.forename: Maurice
authors.initials: M
authors.lastname: Israël
authors.affiliation: Neurobiologie and Développement, Institut de Neurobiologie Alfred Fessard-Fédération pour la Recherche sur le Cerveau (FRC) 2118, Unité Propres de Recherche (UPR) 3294, and.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Benoit
authors.affiliation: Neurobiologie and Développement, Institut de Neurobiologie Alfred Fessard-Fédération pour la Recherche sur le Cerveau (FRC) 2118, Unité Propres de Recherche (UPR) 3294, and sabine.delaporte@inaf.cnrs-gif.fr.
authors.forename: Sabine
authors.initials: S
authors.lastname: de la Porte
mesh_terms: D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000627:therapeutic use; D002087:Butyrates / Q000627:therapeutic use*; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured; D004195:Disease Models, Animal; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006801:Humans; D008297:Male; D051379:Mice; D018101:Mice, Inbred mdx; D018485:Muscle Fibers, Skeletal / Q000187:drug effects; D053580:Muscle Strength / Q000187:drug effects; D009132:Muscles / Q000187:drug effects / Q000502:physiology; D009137:Muscular Dystrophy, Animal / Q000188:drug therapy*; D020388:Muscular Dystrophy, Duchenne / Q000188:drug therapy*; D049411:Utrophin / Q000378:metabolism
chemical_list: D002087:Butyrates; D056572:Histone Deacetylase Inhibitors; D049411:Utrophin; D001120:Arginine; C033018:arginine butyrate
keywords: calcium homeostasis; mass spectrometry imaging; mdx; neuromuscular excitability