Polytherapy with a combination of three repurposed drugs (PXT3003) down-regulates Pmp22 over-expression and improves myelination, axonal and functional parameters in models of CMT1A neuropathy

Orphanet Journal of Rare Diseases

Daniel Cohen¹  Serguei Nabirotchkin¹  Rodolphe Hajj¹  Mickaël Guedj¹  Emmanuel Vial¹  Michael W Sereda²  Klaus-Armin Nave³  Viviane Bertrand¹  Fabrice Glibert¹  Julie Foucquier¹  Julien Laffaire¹  Jonas Mandel¹  Esther Graudens¹  Yannick Pereira¹  Aurélie Boucard¹  Gwenaël Primas¹  Nathalie Cholet¹  Aude Milet¹  Ilya Chumakov¹ 

[1] Pharnext, 11, rue des Peupliers, Issy-Les-Moulineaux, 92130, France;Department of Neurophysiology, University Medical Center, Robert Koch Str. 40, Göttingen, 37075, Germany;Department of Neurogenetics, Max Planck Institute of Experimental Medicine, Hermann-Rein-Strasse 3, Göttingen, 37075, Germany

关键词: Low dose;    CMT1A;    Synergy;    Sorbitol;    Naltrexone;    Baclofen;    Combination therapy;    Repurposing;    Systems Biology;   

Others  :  1138699
DOI  :  10.1186/s13023-014-0201-x

 received in 2014-09-09, accepted in 2014-11-24,  发布年份 2014

【 摘 要 】

Charcot-Marie-Tooth disease type 1A (CMT1A) is the most common inherited sensory and motor peripheral neuropathy. It is caused by PMP22 overexpression which leads to defects of peripheral myelination, loss of long axons, and progressive impairment then disability. There is no treatment available despite observations that monotherapeutic interventions slow progression in rodent models. We thus hypothesized that a polytherapeutic approach using several drugs, previously approved for other diseases, could be beneficial by simultaneously targeting PMP22 and pathways important for myelination and axonal integrity. A combination of drugs for CMT1A polytherapy was chosen from a group of authorised drugs for unrelated diseases using a systems biology approach, followed by pharmacological safety considerations. Testing and proof of synergism of these drugs were performed in a co-culture model of DRG neurons and Schwann cells derived from a Pmp22 transgenic rat model of CMT1A. Their ability to lower Pmp22 mRNA in Schwann cells relative to house-keeping genes or to a second myelin transcript (Mpz) was assessed in a clonal cell line expressing these genes. Finally in vivo efficacy of the combination was tested in two models: CMT1A transgenic rats, and mice that recover from a nerve crush injury, a model to assess neuroprotection and regeneration. Combination of (RS)-baclofen, naltrexone hydrochloride and D-sorbitol, termed PXT3003, improved myelination in the Pmp22 transgenic co-culture cellular model, and moderately down-regulated Pmp22 mRNA expression in Schwannoma cells. In both in vitro systems, the combination of drugs was revealed to possess synergistic effects, which provided the rationale for in vivo clinical testing of rodent models. In Pmp22 transgenic CMT1A rats, PXT3003 down-regulated the Pmp22 to Mpz mRNA ratio, improved myelination of small fibres, increased nerve conduction and ameliorated the clinical phenotype. PXT3003 also improved axonal regeneration and remyelination in the murine nerve crush model. Based on these observations in preclinical models, a clinical trial of PTX3003 in CMT1A, a neglected orphan disease, is warranted. If the efficacy of PTX3003 is confirmed, rational polytherapy based on novel combinations of existing non-toxic drugs with pleiotropic effects may represent a promising approach for rapid drug development.

【 授权许可】

   
2014 Chumakov et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Patzkó A, Shy ME: Update on Charcot-Marie-Tooth disease. Curr Neurol Neurosci Rep 2011, 11:78-88.
• [2]Rossor AM, Polke JM, Houlden H, Reilly MM: Clinical implications of genetic advances in Charcot-Marie-Tooth disease. Nat Rev Neurol 2013, 9:562-571.
• [3]Fledrich R, Stassart RM, Klink A, Rasch LM, Prukop T, Haag L, Czesnik D, Kungl T, Abdelaal TAM, Keric N, Stadelmann C, Brück W, Nave K-A, Sereda MW: Soluble neuregulin-1 modulates disease pathogenesis in rodent models of Charcot-Marie-Tooth disease 1A. Nat Med 2014, 20:1055-1061.
• [4]Nave K-A, Trapp BD: Axon-glial signaling and the glial support of axon function. Annu Rev Neurosci 2008, 31:535-561.
• [5]Sereda M, Griffiths I, Pühlhofer A, Stewart H, Rossner MJ, Zimmerman F, Magyar JP, Schneider A, Hund E, Meinck HM, Suter U, Nave KA: A transgenic rat model of Charcot-Marie-Tooth disease. Neuron 1996, 16:1049-1060.
• [6]Cosgaya JM, Chan JR, Shooter EM: The neurotrophin receptor p75NTR as a positive modulator of myelination. Science 2002, 298:1245-1248.
• [7]Rangaraju S, Madorsky I, Pileggi JG, Kamal A, Notterpek L: Pharmacological induction of the heat shock response improves myelination in a neuropathic model. Neurobiol Dis 2008, 32:105-115.
• [8]Callizot N, Combes M, Steinschneider R, Poindron P: A new long term in vitro model of myelination. Exp Cell Res 2011, 317:2374-2383.
• [9]Sereda MW, Meyer zu Hörste G, Suter U, Uzma N, Nave K-A: Therapeutic administration of progesterone antagonist in a model of Charcot-Marie-Tooth disease (CMT-1A). Nat Med 2003, 9:1533-1537.
• [10]Tashiro H, Fukuda Y, Kimura A, Hoshino S, Ito H, Dohi K: Assessment of microchimerism in rat liver transplantation by polymerase chain reaction. Hepatology 1996, 23:828-834.
• [11]Hirst JA, Howick J, Aronson JK, Roberts N, Perera R, Koshiaris C, Heneghan C: The need for randomization in animal trials: an overview of systematic reviews. PLoS One 2014, 9:e98856.
• [12]Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman DG: Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. J Pharmacol Pharmacother 2010, 1:94-99.
• [13]Meyer Zu Horste G, Prukop T, Liebetanz D, Mobius W, Nave K-A, Sereda MW: Antiprogesterone therapy uncouples axonal loss from demyelination in a transgenic rat model of CMT1A neuropathy. Ann Neurol 2007, 61:61-72.
• [14]Rivlin AS, Tator CH: Objective clinical assessment of motor function after experimental spinal cord injury in the rat. J Neurosurg 1977, 47:577-581.
• [15]Beyreuther BK, Callizot N, Brot MD, Feldman R, Bain SC, Stöhr T: Antinociceptive efficacy of lacosamide in rat models for tumor- and chemotherapy-induced cancer pain. Eur J Pharmacol 2007, 565:98-104.
• [16]Tétreault P, Dansereau M-A, Doré-Savard L, Beaudet N, Sarret P: Weight bearing evaluation in inflammatory, neuropathic and cancer chronic pain in freely moving rats. Physiol Behav 2011, 104:495-502.
• [17]Bordet T, Buisson B, Michaud M, Drouot C, Gale P, Delaage P, Akentieva NP, Evers AS, Covey DF, Ostuni MA, Lacape J, Massaad C, Schumacher M, Steidl E, Maux D, Delaage M, Henderson CE, Pruss RM: Identification and characterization of cholest-4-en-3-one, oxime (TRO19622), a novel drug candidate for amyotrophic lateral sclerosis. J Pharmacol Exp Ther 2007, 322:709-720.
• [18]Markowski C, Markowski E: Conditions for the effectiveness of a preliminary test of variance. Am Stat 1990, 44:322-326.
• [19]Sawilowsky SS, Blair RC: A more realistic look at the robustness and Type II error properties of the t test to departures from population normality. Psychol Bull 1992, 111:352-360.
• [20]Grabovsky Y, Tallarida RJ: Isobolographic analysis for combinations of a full and partial agonist: curved isoboles. J Pharmacol Exp Ther 2004, 310:981-986.
• [21]Chou T-C: Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 2006, 58:621-681.
• [22]Geary N: Understanding synergy. Am J Physiol Metab 2013, 304:E237-E253.
• [23]Tallarida RJ: An overview of drug combination analysis with isobolograms. J Pharmacol Exp Ther 2006, 319:1-7.
• [24]Suter U, Snipes GJ, Schoener-Scott R, Welcher AA, Pareek S, Lupski JR, Murphy RA, Shooter EM, Patel PI: Regulation of tissue-specific expression of alternative peripheral myelin protein-22 (PMP22) gene transcripts by two promoters. J Biol Chem 1994, 269:25795-25808.
• [25]Sabéran-Djoneidi D, Sanguedolce V, Assouline Z, Lévy N, Passage E, Fontés M: Molecular dissection of the Schwann cell specific promoter of the PMP22 gene. Gene 2000, 248:223-231.
• [26]Ogata T, Iijima S, Hoshikawa S, Miura T, Yamamoto S, Oda H, Nakamura K, Tanaka S: Opposing extracellular signal-regulated kinase and Akt pathways control Schwann cell myelination. J Neurosci 2004, 24:6724-6732.
• [27]Arthur-Farraj P, Wanek K, Hantke J, Davis CM, Jayakar A, Parkinson DB, Mirsky R, Jessen KR: Mouse schwann cells need both NRG1 and cyclic AMP to myelinate. Glia 2011, 59:720-733.
• [28]Pereira JA, Lebrun-Julien F, Suter U: Molecular mechanisms regulating myelination in the peripheral nervous system. Trends Neurosci 2012, 35:123-134.
• [29]Faroni A, Magnaghi V: The neurosteroid allopregnanolone modulates specific functions in central and peripheral glial cells. Front Endocrinol (Lausanne) 2011, 2:103.
• [30]Procacci P, Ballabio M, Castelnovo LF, Mantovani C, Magnaghi V: GABA-B receptors in the PNS have a role in Schwann cells differentiation? Front Cell Neurosci 2012, 6:68.
• [31]Glenn TD, Talbot WS: Signals regulating myelination in peripheral nerves and the Schwann cell response to injury. Curr Opin Neurobiol 2013, 23:1041-1048.
• [32]Towers S, Princivalle A, Billinton A, Edmunds M, Bettler B, Urban L, Bowery NG: GABA B receptor protein and mRNA distribution in rat spinal cord and dorsal root ganglia. Eur J Neurosci 2000, 12:3201-3210.
• [33]Wang H-Y, Frankfurt M, Burns LH: High-affinity naloxone binding to filamin a prevents mu opioid receptor-Gs coupling underlying opioid tolerance and dependence. PLoS One 2008, 3:e1554.
• [34]Hytrek SD, McLaughlin PJ, Lang CM, Zagon IS: Inhibition of human colon cancer by intermittent opioid receptor blockade with naltrexone. Cancer Lett 1996, 101:159-164.
• [35]Zhu M, Li RC: Receptor binding activities of Schefflera triterpenoids and oligosaccharides. Planta Med 1999, 65:99-103.
• [36]Singer MA, Lindquist S: Multiple effects of trehalose on protein folding in vitro and in vivo. Mol Cell 1998, 1:639-648.
• [37]Kumar R: Role of naturally occurring osmolytes in protein folding and stability. Arch Biochem Biophys 2009, 491:1-6.
• [38]Notterpek L, Ryan MC, Tobler AR, Shooter EM: PMP22 accumulation in aggresomes: implications for CMT1A pathology. Neurobiol Dis 1999, 6:450-460.
• [39]Schlebach JP, Peng D, Kroncke BM, Mittendorf KF, Narayan M, Carter BD, Sanders CR: Reversible folding of human peripheral myelin protein 22, a tetraspan membrane protein. Biochemistry 2013, 52:3229-3241.
• [40]Nobbio L, Mancardi G, Grandis M, Levi G, Suter U, Nave KA, Windebank AJ, Abbruzzese M, Schenone A: PMP22 transgenic dorsal root ganglia cultures show myelin abnormalities similar to those of human CMT1A. Ann Neurol 2001, 50:47-55.
• [41]Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002, 3:RESEARCH0034. BioMed Central Full Text
• [42]Fledrich R, Schlotter-Weigel B, Schnizer TJ, Wichert SP, Stassart RM, Meyer zu Hörste G, Klink A, Weiss BG, Haag U, Walter MC, Rautenstrauss B, Paulus W, Rossner MJ, Sereda MW: A rat model of Charcot-Marie-Tooth disease 1A recapitulates disease variability and supplies biomarkers of axonal loss in patients. Brain 2012, 135:72-87.
• [43]Shy ME, Chen L, Swan ER, Taube R, Krajewski KM, Herrmann D, Lewis RA, McDermott MP: Neuropathy progression in Charcot-Marie-Tooth disease type 1A. Neurology 2008, 70:378-383.
• [44]Pareyson D, Scaioli V, Laurà M: Clinical and electrophysiological aspects of Charcot-Marie-Tooth disease. Neuromolecular Med 2006, 8:3-22.
• [45]Martini R, Klein D, Groh J: Similarities between inherited demyelinating neuropathies and Wallerian degeneration: an old repair program may cause myelin and axon perturbation under nonlesion conditions. Am J Pathol 2013, 183:655-660.
• [46]Castorina A, Scuderi S, D’Amico AG, Drago F, D’Agata V: PACAP and VIP increase the expression of myelin-related proteins in rat schwannoma cells: involvement of PAC1/VPAC2 receptor-mediated activation of PI3K/Akt signaling pathways. Exp Cell Res 2014, 322:108-121.
• [47]Hai M, Muja N, DeVries GH, Quarles RH, Patel PI: Comparative analysis of Schwann cell lines as model systems for myelin gene transcription studies. J Neurosci Res 2002, 69:497-508.
• [48]Nobbio L, Visigalli D, Radice D, Fiorina E, Solari A, Lauria G, Reilly MM, Santoro L, Schenone A, Pareyson D: PMP22 messenger RNA levels in skin biopsies: testing the effectiveness of a Charcot-Marie-Tooth 1A biomarker. Brain 2014, 137:1614-1620.
• [49]Katona I, Wu X, Feely SME, Sottile S, Siskind CE, Miller LJ, Shy ME, Li J: PMP22 expression in dermal nerve myelin from patients with CMT1A. Brain 2009, 132:1734-1740.
• [50]Hay M, Thomas DW, Craighead JL, Economides C, Rosenthal J: Clinical development success rates for investigational drugs. Nat Biotechnol 2014, 32:40-51.
• [51]Attarian S, Vallat J-M, Magy L, Funalot B, Gonnaud P-M, Lacour A, Péréon Y, Dubourg O, Pouget J, Micallef J, Franques J, Lefèvre M-N, Ghorab K, Al-Moussawi M, Tiffreau V, Magot A, Leclair-Visonneau L, Stojkovic T, Bossi L, Lehert P, Walter G, Bertrand V, Mandel J, Milet A, Hajj R, Boudiaf L, Scart-Grès C, Nabirotchkin S, Guedj M, Chumakov I, et al: An Exploratory randomised double-blind and placebo-controlled phase 2 study of a combination of baclofen, naltrexone and sorbitol (PXT3003) in patients with Charcot-Marie-Tooth disease type 1A.Orphanet J Rare Dis 2014, 9:199.