BACKGROUND: Charcot-Marie-Tooth disease type 1A (CMT1A) is the most common inherited neuropathy, a debilitating disease without known cure. Among patients with CMT1A, disease manifestation, progression and severity are strikingly variable, which poses major challenges for the development of new therapies. Hence, there is a strong need for sensitive outcome measures such as disease and progression biomarkers, which would add powerful tools to monitor therapeutic effects in CMT1A. METHODS: We established a pan-European and American consortium comprising nine clinical centres including 311 patients with CMT1A in total. From all patients, the CMT neuropathy score and secondary outcome measures were obtained and a skin biopsy collected. In order to assess and validate disease severity and progression biomarkers, we performed qPCR on a set of 16 animal model-derived potential biomarkers in skin biopsy mRNA extracts. RESULTS: In 266 patients with CMT1A, a cluster of eight cutaneous transcripts differentiates disease severity with a sensitivity and specificity of 90% and 76.1%, respectively. In an additional cohort of 45 patients with CMT1A, from whom a second skin biopsy was taken after 2-3 years, the cutaneous mRNA expression of GSTT2, CTSA, PPARG, CDA, ENPP1 and NRG1-Iis changing over time and correlates with disease progression. CONCLUSIONS: In summary, we provide evidence that cutaneous transcripts in patients with CMT1A serve as disease severity and progression biomarkers and, if implemented into clinical trials, they could markedly accelerate the development of a therapy for CMT1A.

Center for Molecular Neurology VIB Antwerp Belgium

CMT TRIAAL Milan Italy

Department of Child Neurology Charles University 2nd Medical School University Hospital Motol Prague Czech Republic

Department of Clinical Neurophysiology University Medical Center Göttingen Göttingen Germany

Department of Clinical Neurosciences Unit of Neurological Rare Diseases of Adulthood IRCCS Foundation C Besta Neurological Institute Milan Italy

Department of Medical Statistics University Medical Center Göttingen Göttingen Germany

Department of Neurogenetics Max Planck Institute of Experimental Medicine Göttingen Germany

Department of Neurology Antwerp University Hospital Antwerpen Belgium

Department of Neurology Carver College of Medicine University of Iowa Iowa USA

Department of Neurology Ophthalmology and Genetics University of Genoa Genoa Italy

Department of Sleep Medicine and Neuromuscular Diseases University of Münster Münster Germany

Friedrich Baur Institut Department of Neurology Ludwig Maximilians University of Munich Munich Germany

Institute Born Bunge University of Antwerp Antwerpen Belgium

Institute of Clinical Pharmacology University Medical Center Göttingen Göttingen Germany

Institute of Myology GH Pitié Salpêtrière Paris France

John Walton Muscular Dystrophy Research Centre Institute of Genetic Medicine Newcastle University UK

Service of Neurology University Hospital Marqués de Valdecilla Santander Spain

Unit of Neuroepidemiology IRCCS Foundation C Besta Neurological Institute Milan Italy

Zobrazit více v PubMed

Skre H.Genetic and clinical aspects of Charcot-Marie-Tooth’s disease. Clin Genet [Internet]. 1974. January [cited 2013 Oct 1];6(2):98–118. Available from: http://www.ncbi.nlm.nih.gov/pubmed/4430158 PubMed

Braathen GJ, Sand JC, Lobato a, Høyer H, Russell MB. Genetic epidemiology of Charcot-Marie-Tooth in the general population. Eur J Neurol Off J Eur Fed Neurol Soc [Internet]. 2011. January [cited 2013 Mar 8];18(1):39–48. Available from: http://www.ncbi.nlm.nih.gov/pubmed/20482598 PubMed

Timmerman V, Strickland A V, Züchner S. Genetics of Charcot-Marie-Tooth (CMT) Disease within the Frame of the Human Genome Project Success. 2014;13–32. PubMed PMC

Lupski JR, De Oca-Luna RM, Slaugenhaupt S, Pentao L, Guzzetta V, Trask BJ, et al. DNA duplication associated with Charcot-Marie-Tooth disease type 1A. Cell [Internet]. 1991;66(2):219–32. Available from: http://www.ncbi.nlm.nih.gov/pubmed/1677316 PubMed

Raeymaekers P, Timmerman V, Nelis E, De Jonghe P, Hoogendijk JE, Baas F, et al. Duplication in chromosome 17p11.2 in Charcot-Marie-Tooth neuropathy type 1a (CMT 1a). The HMSN Collaborative Research Group. Neuromuscul Disord [Internet]. 1991. January [cited 2013 Oct 1];1(2):93–7. Available from: http://www.ncbi.nlm.nih.gov/pubmed/1822787 PubMed

Matsunami N, Smith B, Ballard L, Lensch MW, Robertson M, Albertsen H, et al. Peripheral myelin protein-22 gene maps in the duplication in chromosome 17p11.2 associated with Charcot-Marie-Tooth 1A. Nat Genet [Internet]. 1992. June [cited 2014 Feb 12];1(3):176–9. Available from: http://www.ncbi.nlm.nih.gov/pubmed/1303231 PubMed

Patel PI, Roa BB, Welcher AA, Schoener-Scott R, Trask BJ, Pentao L, et al. The gene for the peripheral myelin protein PMP-22 is a candidate for Charcot-Marie-Tooth disease type 1A. Nat Genet. 1992;1(3):159–65. PubMed

Timmerman V, Nelis E, Van Hul W, Nieuwenhuijsen BW, Chen KL, Wang S, et al. The peripheral myelin protein gene PMP-22 is contained within the Charcot-Marie-Tooth disease type 1A duplication. Nat Genet [Internet]. 1992. June [cited 2014 Feb 12];1(3):171–5. Available from: http://www.ncbi.nlm.nih.gov/pubmed/1303230 PubMed

Valentijn LJ, Baas F, Wolterman RA, Hoogendijk JE, van den Bosch NH, Zorn I, et al. Identical point mutations of PMP-22 in Trembler-J mouse and Charcot-Marie-Tooth disease type 1A. Nat Genet [Internet]. 1992. December [cited 2013 Oct 1];2(4):288–91. Available from: http://www.ncbi.nlm.nih.gov/pubmed/1303281 PubMed

Dyck PJ, Thomas PK. Peripheral Neuropathy, 4th Edition | P. K. Thomas, Peter Dyck; | ISBN 9780721694917 [Internet]. Book. 2005. [cited 2013 Sep 16]. Available from: http://store.elsevier.com/Peripheral-Neuropathy/P_-K_-Thomas/isbn-9780721694917/

Harding A, Thomas P. The clinical features of hereditary motor and sensory neuropathy types I and II. Brain a J Neurol [Internet]. 1980. [cited 2013 Oct 1];17(2):259–80. Available from: http://www.ncbi.nlm.nih.gov/pubmed/7397478 PubMed

Yiu EM, Burns J, Ryan MM, Ouvrier RA. Neurophysiologic abnormalities in children with Charcot-Marie-Tooth disease type 1A. J Peripher Nerv Syst [Internet]. 2008. September [cited 2014 Feb 13];13(3):236–41. Available from: http://www.ncbi.nlm.nih.gov/pubmed/18844790 PubMed

Kaku D, Parry G, Malamut R, Lupski J, Garcia C. Uniform slowing of conduction velocities in Charcot‐Marie‐Tooth polyneuropathy type 1. Neurology [Internet]. 1993. [cited 2013 Oct 1]; Available from: http://www.neurology.org/content/43/12/2664.short PubMed

Garcia CA, Malamut RE, England JD, Parry GS, Liu P, Lupski JR. Clinical variability in two pairs of identical twins with the Charcot-Marie-Tooth disease type 1A duplication. Neurology [Internet]. 1995. November [cited 2013 Oct 1];45(11):2090–3. Available from: http://www.ncbi.nlm.nih.gov/pubmed/7501164 PubMed

Reilly MM, Shy ME, Muntoni F, Pareyson D. 168th ENMC International workshop: Outcome measures and clinical trials in Charcot-Marie-Tooth disease (CMT). Neuromuscul Disord [Internet]. 2010;20(12):839–46. Available from: 10.1016/j.nmd.2010.08.001 PubMed DOI

Fledrich R, Schlotter-Weigel B, Schnizer TJ, Wichert SP, Stassart RM, Zu Hörste GM, et al. A rat model of Charcot-Marie-Tooth disease 1A recapitulates disease variability and supplies biomarkers of axonal loss in patients. Brain A J Neurol [Internet]. 2012;135(Pt 1):72–87. Available from: http://www.ncbi.nlm.nih.gov/pubmed/22189569 PubMed

Young P, De Jonghe P, Stögbauer F, Butterfass-Bahloul T. Treatment for Charcot-Marie-Tooth disease. Cochrane database Syst Rev [Internet]. 2008. January [cited 2013 Oct 1];(1):CD006052. Available from: http://www.ncbi.nlm.nih.gov/pubmed/18254090 PubMed PMC

Burns J, Ouvrier RA, Yiu EM, Joseph PD, Kornberg AJ, Fahey MC, et al. Ascorbic acid for Charcot-Marie-Tooth disease type 1A in children: a randomised, double-blind, placebo-controlled, safety and efficacy trial. Lancet Neurol [Internet]. 2009. June [cited 2013 Oct 1];8(6):537–44. Available from: http://www.ncbi.nlm.nih.gov/pubmed/19427269 PubMed

Micallef J, Attarian S, Dubourg O, Gonnaud P-M, Hogrel J-Y, Stojkovic T, et al. Effect of ascorbic acid in patients with Charcot-Marie-Tooth disease type 1A: a multicentre, randomised, double-blind, placebo-controlled trial. Lancet Neurol [Internet]. 2009. December [cited 2013 Oct 1];8(12):1103–10. Available from: http://www.ncbi.nlm.nih.gov/pubmed/19818690 PubMed

Verhamme C, de Haan RJ, Vermeulen M, Baas F, de Visser M, van Schaik IN. Oral high dose ascorbic acid treatment for one year in young CMT1A patients: a randomised, double-blind, placebo-controlled phase II trial. BMC Med [Internet]. 2009. January [cited 2013 Oct 1];7:70. Available from: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2784478&tool=pmcentrez&rendertype=abstract PubMed PMC

Pareyson D, Reilly MM, Schenone A, Fabrizi GM, Cavallaro T, Santoro L, et al. Ascorbic acid in Charcot–Marie–Tooth disease type 1A (CMT-TRIAAL and CMT-TRAUK): a double-blind randomised trial. Lancet [Internet]. 2011. April [cited 2013 Mar 8];10(4):320–8. Available from: http://discovery.ucl.ac.uk/1304687/ PubMed PMC

de Visser M, Verhamme C. Ascorbic acid for treatment in CMT1A: What’s next? Lancet Neurol [Internet]. 2011;10(4):291–3. Available from: 10.1016/S1474-4422(11)70042-4 PubMed DOI

Murphy SM, Herrmann DN, McDermott MP, Scherer SS, Shy ME, Reilly MM, et al. Reliability of the CMT neuropathy score (second version) in Charcot-Marie-Tooth disease. J Peripher Nerv Syst. 2011;16(3):191–8. PubMed PMC

Mannil M, Solari A, Leha A, Pelayo-Negro AL, Berciano J, Schlotter-Weigel B, et al. Selected items from the Charcot-Marie-Tooth (CMT) Neuropathy Score and secondary clinical outcome measures serve as sensitive clinical markers of disease severity in CMT1A patients. Neuromuscul Disord. 2014;24(11):1003–17. PubMed

Shy ME, Blake J, Krajewski K, Fuerst DR, Laura M, Hahn a F, et al. Reliability and validity of the CMT neuropathy score as a measure of disability. Neurology [Internet]. 2005. April 12 [cited 2013 Mar 8];66(7):614–615-615. Available from: http://discovery.ucl.ac.uk/157383/ PubMed

Sadjadi R, Reilly MM, Shy ME, Pareyson D, Laura M, Murphy S, et al. Psychometrics evaluation of Charcot-Marie-Tooth Neuropathy Score (CMTNSv2 ) second version, using Rasch analysis. 2014;196:192–6. PubMed PMC

Fledrich R, Stassart RM, Sereda MW. Murine therapeutic models for Charcot-Marie-Tooth (CMT) disease. Br Med Bull [Internet]. 2012. June [cited 2013 Mar 8];102:89–113. Available from: http://www.ncbi.nlm.nih.gov/pubmed/22551516 PubMed

Willcocks RJ, Rooney WD, Triplett WT, Forbes SC, Lott DJ, Senesac CR, et al. Multicenter Prospective Longitudinal Study of Magnetic Resonance Biomarkers in a Large Duchenne Muscular Dystrophy Cohort. 2016; PubMed PMC

Filler AG, Maravilla KR, Tsuruda JS. MR neurography and muscle MR imaging for image diagnosis of disorders affecting the peripheral nerves and musculature. 2004;22:8619. PubMed

Morrow JM, Sinclair CDJ, Fischmann A, Machado PM, Reilly MM, Yousry TA, et al. MRI biomarker assessment of neuromuscular disease progression : a prospective observational cohort study. Lancet Neurol [Internet]. 2016;15(1):65–77. Available from: 10.1016/S1474-4422(15)00242-2 PubMed DOI PMC

Dortch RD, Dethrage LM, Gore JC, Smith SA. Proximal nerve magnetization transfer MRI relates to disability in Charcot-Marie-Tooth diseases. 2014;1545–54. PubMed PMC

Gramont A de, Watson S, Ellis LM, Rodón J, Tabernero J, Gramont A de, et al. Pragmatic issues in biomarker evaluation for targeted therapies in cancer. Nat Rev Clin Oncol. 2015;12:197–2012. PubMed

Lee JW, Devanarayan V, Barrett YC, Weiner R, Allinson J, Fountain S, et al. Fit-for-Purpose Method Development and Validation for Successful Biomarker Measurement. 2006;23(2):312–28. PubMed

Sereda M, Griffiths I, Pühlhofer a, Stewart H, Rossner MJ, Zimmerman F, et al. A transgenic rat model of Charcot-Marie-Tooth disease. Neuron [Internet]. 1996;16(5):1049–60. Available from: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8630243 PubMed

Kuleshov M V, Jones MR, Rouillard AD, Fernandez NF, Duan Q, Wang Z, et al. Enrichr: a comprehensive gene set enrichment analysis web server 2016 update. Nucleic Acids Res [Internet]. 2016;44(W1):W90–7. Available from: http://www.ncbi.nlm.nih.gov/pubmed/27141961 PubMed PMC

Shy ME, Chen L, Swan ER, Taube R, Krajewski KM, Herrmann D, et al. Neuropathy progression in Charcot-Marie-Tooth disease type 1A. Cancer [Internet]. 2008. January 29 [cited 2013 Mar 8];77(7):1356–62. Available from: http://www.ncbi.nlm.nih.gov/pubmed/18227419 PubMed

Berciano J, Gallardo E, Garcia A, Ramón C, Infante J, Combarros O. Clinical progression in Charcot – Marie-Tooth disease type 1A duplication : clinico-electrophysiological and MRI longitudinal study of a family. J Neurol. 2010;257:1633–41. PubMed

Piscosquito G, Reilly MM, Schenone A, Fabrizi GM, Cavallaro T, Santoro L, et al. Responsiveness of clinical outcome measures in Charcot À Marie À Tooth disease. 2015;1556–63. PubMed

Manganelli F, Stancanelli A, Caporaso G, Shy ME. Charcot-Marie-Tooth disease. 2015; PubMed PMC

Fledrich R, Schlotter-Weigel B, Schnizer TJ, Wichert SP, Stassart RM, Zu Hörste GM, et al. A rat model of Charcot-Marie-Tooth disease 1A recapitulates disease variability and supplies biomarkers of axonal loss in patients. Brain A J Neurol [Internet]. 2012. January [cited 2013 Mar 8];135(Pt 1):72–87. Available from: http://www.ncbi.nlm.nih.gov/pubmed/22189569 PubMed

Dacci P, Dina G, Cerri F, Previtali SC, Lopez ID, Lauria G, et al. Foot Pad Skin Biopsy in Mouse Models of Hereditary Neuropathy. Glia [Internet]. 2010. December [cited 2013 Mar 8];58(16):2005–16. Available from: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3034192&tool=pmcentrez&rendertype=abstract PubMed PMC

Fortun J, Go JC, Li J, Amici S a, Dunn W a, Notterpek L. Alterations in degradative pathways and protein aggregation in a neuropathy model based on PMP22 overexpression. Neurobiol Dis [Internet]. 2006. April [cited 2013 Mar 8];22(1):153–64. Available from: http://www.ncbi.nlm.nih.gov/pubmed/16326107 PubMed