title: Clobetasol and Halcinonide Act as Smoothened Agonists to Promote Myelin Gene Expression and RxRγ Receptor Activation.
abstract: One of the causes of permanent disability in chronic multiple sclerosis patients is the inability of oligodendrocyte progenitor cells (OPCs) to terminate their maturation program at lesions. To identify key regulators of myelin gene expression acting at the last stages of OPC maturation we developed a drug repositioning strategy based on the mouse immortalized oligodendrocyte (OL) cell line Oli-neu brought to the premyelination stage by stably expressing a key factor regulating the last stages of OL maturation. The Prestwick Chemical Library of 1,200 FDA-approved compound(s) was repositioned at three dosages based on the induction of Myelin Basic Protein (MBP) expression. Drug hits were further validated using dosage-dependent reproducibility tests and biochemical assays. The glucocorticoid class of compounds was the most highly represented and we found that they can be divided in three groups according to their efficacy on MBP up-regulation. Since target identification is crucial before bringing compounds to the clinic, we searched for common targets of the primary screen hits based on their known chemical-target interactomes, and the pathways predicted by top ranking compounds were validated using specific inhibitors. Two of the top ranking compounds, Halcinonide and Clobetasol, act as Smoothened (Smo) agonists to up-regulate myelin gene expression in the Oli-neuM cell line. Further, RxRγ activation is required for MBP expression upon Halcinonide and Clobetasol treatment. These data indicate Clobetasol and Halcinonide as potential promyelinating drugs and also provide a mechanistic understanding of their mode of action in the pathway leading to myelination in OPCs. Furthermore, our classification of glucocorticoids with respect to MBP expression provides important novel insights into their effects in the CNS and a rational criteria for their choice in combinatorial therapies in de-myelinating diseases.
authors.affiliation: Department of Biology, University of Rome "Tor Vergata", Rome, Italy.|Department of Translational Pharmacology, Fondazione Mario Negri Sud, S. Maria Imbaro (CH), Italy.
authors.forename: Giampiero
authors.initials: G
authors.lastname: Porcu
authors.affiliation: Department of Translational Pharmacology, Fondazione Mario Negri Sud, S. Maria Imbaro (CH), Italy.|Department of Life, Health and Environmental Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Eliseo
authors.initials: E
authors.lastname: Serone
authors.affiliation: Department of Translational Pharmacology, Fondazione Mario Negri Sud, S. Maria Imbaro (CH), Italy.
authors.forename: Velia
authors.initials: V
authors.lastname: De Nardis
authors.affiliation: Department of Translational Pharmacology, Fondazione Mario Negri Sud, S. Maria Imbaro (CH), Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Di Giandomenico
authors.affiliation: Department of Translational Pharmacology, Fondazione Mario Negri Sud, S. Maria Imbaro (CH), Italy.|Center for Outcomes Research and Clinical Epidemiology, Pescara, Italy.|Dipartimento di Scienze Mediche di Base, Neuroscienze ed Organi di Senso, Università di Bari Aldo Moro, Bari, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Lucisano
authors.affiliation: Department of Translational Pharmacology, Fondazione Mario Negri Sud, S. Maria Imbaro (CH), Italy.|Center for Outcomes Research and Clinical Epidemiology, Pescara, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Scardapane
authors.affiliation: Department of Life, Health and Environmental Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Poma
authors.affiliation: Department of Biology, University of Rome "Tor Vergata", Rome, Italy.|Department of Translational Pharmacology, Fondazione Mario Negri Sud, S. Maria Imbaro (CH), Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Ragnini-Wilson
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002460:Cell Line; D002990:Clobetasol / Q000494:pharmacology*; D003598:Cytoskeletal Proteins / Q000819:agonists / Q000378:metabolism*; D058492:Drug Repositioning; D015870:Gene Expression / Q000187:drug effects; D006206:Halcinonide / Q000494:pharmacology*; D015151:Immunoblotting; D051379:Mice; D008856:Microscopy, Fluorescence; D009124:Muscle Proteins / Q000819:agonists / Q000378:metabolism*; D004676:Myelin Basic Protein / Q000378:metabolism; D009186:Myelin Sheath / Q000378:metabolism*; D009836:Oligodendroglia / Q000187:drug effects / Q000378:metabolism; D047491:Retinoid X Receptor gamma / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics
chemical_list: D000893:Anti-Inflammatory Agents; D003598:Cytoskeletal Proteins; D009124:Muscle Proteins; D004676:Myelin Basic Protein; D047491:Retinoid X Receptor gamma; C491050:Smtn protein, mouse; D014157:Transcription Factors; C571125:myelin gene regulatory factor, mouse; D002990:Clobetasol; D006206:Halcinonide