Neurobiology of Aging 35 (2014) 1213.e1e1213.e2
Contents lists available at ScienceDirect
Neurobiology of Aging journal homepage: www.elsevier.com/locate/neuaging
Negative results
C9ORF72 repeat expansion not detected in patients with multiple sclerosis Chiara Fenoglio, Milena De Riz, Chiara Villa, Maria Serpente, Elisa Ridolfi, Rossana Bonsi, Sara M.G. Cioffi, Cinzia Barone, Anna Pietroboni, Alberto Calvi, Elio Scarpini, Daniela Galimberti* Neurology Unit, Department of Pathophysiology and Transplantation, University of Milan, “Dino Ferrari” Center, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, Milan, Italy
a r t i c l e i n f o
a b s t r a c t
Article history: Received 25 October 2013 Accepted 28 October 2013 Available online 1 November 2013
A hexanucleotide repeat expansion in the chromosome 9 Open Reading Frame 72 gene (C9ORF72) has recently been reported to be cause of familial amyotrophic lateral sclerosis and frontotemporal lobar degeneration. Nevertheless, in the last few years this mutation has been found to be associated with heterogeneous phenotypes, including multiple sclerosis (MS) in concurrence with amyotrophic lateral sclerosis. In this study, we sought to evaluate the presence of the C9ORF72 repeat expansion in a cohort consisting of 314 patients with MS and 222 control subjects. No pathogenic expansion was found in MS and control populations, suggesting that C9ORF72 does not play a major role in MS pathogenesis. Ó 2014 Elsevier Inc. All rights reserved.
Keywords: Multiple sclerosis C9ORF72 repeat expansion Neurodegeneration
1. Introduction
2. Methods
A hexanucleotide repeat expansion in chromosome 9 Open Reading Frame 72 gene (C9ORF72) has recently been identified in familial amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (DeJesus-Hernandez et al., 2011; Renton et al., 2011). Nevertheless, the mutation is associated with a wide variety of clinical phenotypes (Galimberti et al., 2013; Xi et al., 2012). Ismail et al. identified the mutation in 5 patients with multiple sclerosis (MS)-ALS. These patients had a more rapidly progressive disease than pure C9ORF72-ALS (Ismail et al., 2012). Multiple sclerosis may be associated with cognitive impairment (Lovera and Kovner, 2012) and besides typical white matter involvement, gray matter damage have been recently described (Geurts et al., 2013). In light of these findings, we screened a cohort of 314 patients with MS, and 222 control subjects, for the presence of the C9ORF72 repeat expansion, to determine a possible role of this mutation in MS pathogenesis.
Patients (96 males and 218 females) were consecutively recruited at the Multiple Sclerosis Center of the Fondazione Cà Granda IRCCS Ospedale Maggiore Policlinico. Diagnosis was done in accordance with current criteria (Polman et al., 2005). In addition, we analyzed 222 healthy age-matched control subjects (92 males and 130 females). Characteristics of the patients and control subjects are summarized in the Supplementary Table 1. High-molecular weight DNA was isolated from whole blood using the Flexigene Kit (Qiagen, Hildren, Germany) and C9ORF72 genotyping carried out by repeat-primed polymerase chain reaction and sequencing, as previously described (DeJesus-Hernandez et al., 2011). A characteristic stutter amplification pattern (>30 repeats) on the electropherogram was considered evidence of a pathogenic repeat expansion (Dobson-Stone et al., 2012).
* Corresponding author at: Neurology Unit, Department of Pathophysiology and Transplantation, University of Milan, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, via F. Sforza, 35, 20122, Milan, Italy. Tel.: þ39 02 55033847; fax: þ39 02 55036580. E-mail address: [email protected] (D. Galimberti).
4. Discussion
0197-4580/$ e see front matter Ó 2014 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.neurobiolaging.2013.10.096
3. Results The pathogenic C9ORF72 expansion was not present in the DNA of any patient or control subject. Maximum length detected in both patients and control subjects was 15 repeats.
Herein, we showed that the C9ORF72 mutation is not associated with MS, suggesting that C9ORF72 repeat expansion do not
1213.e2
C. Fenoglio et al. / Neurobiology of Aging 35 (2014) 1213.e1e1213.e2
play a major role in the pathogenesis of the disease. Previous data (Ismail et al., 2012) demonstrated the presence of the mutation in patients with MS and ALS, but not MS alone. Taken together, these findings suggest that the expansion could be causative of ALS independent of the co-occurrence of MS.
Acknowledgements This work was supported by grant Italian Ministry of Health (Ricerca Corrente) and Fondazione Monzino. Appendix A. Supplementary data
Disclosure statement The authors have no actual or potential conflicts of interest.
Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j.neurobiolaging. 2013.10.096.
Contents lists available at ScienceDirect
Neurobiology of Aging journal homepage: www.elsevier.com/locate/neuaging
Negative results
C9ORF72 repeat expansion not detected in patients with multiple sclerosis Chiara Fenoglio, Milena De Riz, Chiara Villa, Maria Serpente, Elisa Ridolfi, Rossana Bonsi, Sara M.G. Cioffi, Cinzia Barone, Anna Pietroboni, Alberto Calvi, Elio Scarpini, Daniela Galimberti* Neurology Unit, Department of Pathophysiology and Transplantation, University of Milan, “Dino Ferrari” Center, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, Milan, Italy
a r t i c l e i n f o
a b s t r a c t
Article history: Received 25 October 2013 Accepted 28 October 2013 Available online 1 November 2013
A hexanucleotide repeat expansion in the chromosome 9 Open Reading Frame 72 gene (C9ORF72) has recently been reported to be cause of familial amyotrophic lateral sclerosis and frontotemporal lobar degeneration. Nevertheless, in the last few years this mutation has been found to be associated with heterogeneous phenotypes, including multiple sclerosis (MS) in concurrence with amyotrophic lateral sclerosis. In this study, we sought to evaluate the presence of the C9ORF72 repeat expansion in a cohort consisting of 314 patients with MS and 222 control subjects. No pathogenic expansion was found in MS and control populations, suggesting that C9ORF72 does not play a major role in MS pathogenesis. Ó 2014 Elsevier Inc. All rights reserved.
Keywords: Multiple sclerosis C9ORF72 repeat expansion Neurodegeneration
1. Introduction
2. Methods
A hexanucleotide repeat expansion in chromosome 9 Open Reading Frame 72 gene (C9ORF72) has recently been identified in familial amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (DeJesus-Hernandez et al., 2011; Renton et al., 2011). Nevertheless, the mutation is associated with a wide variety of clinical phenotypes (Galimberti et al., 2013; Xi et al., 2012). Ismail et al. identified the mutation in 5 patients with multiple sclerosis (MS)-ALS. These patients had a more rapidly progressive disease than pure C9ORF72-ALS (Ismail et al., 2012). Multiple sclerosis may be associated with cognitive impairment (Lovera and Kovner, 2012) and besides typical white matter involvement, gray matter damage have been recently described (Geurts et al., 2013). In light of these findings, we screened a cohort of 314 patients with MS, and 222 control subjects, for the presence of the C9ORF72 repeat expansion, to determine a possible role of this mutation in MS pathogenesis.
Patients (96 males and 218 females) were consecutively recruited at the Multiple Sclerosis Center of the Fondazione Cà Granda IRCCS Ospedale Maggiore Policlinico. Diagnosis was done in accordance with current criteria (Polman et al., 2005). In addition, we analyzed 222 healthy age-matched control subjects (92 males and 130 females). Characteristics of the patients and control subjects are summarized in the Supplementary Table 1. High-molecular weight DNA was isolated from whole blood using the Flexigene Kit (Qiagen, Hildren, Germany) and C9ORF72 genotyping carried out by repeat-primed polymerase chain reaction and sequencing, as previously described (DeJesus-Hernandez et al., 2011). A characteristic stutter amplification pattern (>30 repeats) on the electropherogram was considered evidence of a pathogenic repeat expansion (Dobson-Stone et al., 2012).
* Corresponding author at: Neurology Unit, Department of Pathophysiology and Transplantation, University of Milan, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, via F. Sforza, 35, 20122, Milan, Italy. Tel.: þ39 02 55033847; fax: þ39 02 55036580. E-mail address: [email protected] (D. Galimberti).
4. Discussion
0197-4580/$ e see front matter Ó 2014 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.neurobiolaging.2013.10.096
3. Results The pathogenic C9ORF72 expansion was not present in the DNA of any patient or control subject. Maximum length detected in both patients and control subjects was 15 repeats.
Herein, we showed that the C9ORF72 mutation is not associated with MS, suggesting that C9ORF72 repeat expansion do not
1213.e2
C. Fenoglio et al. / Neurobiology of Aging 35 (2014) 1213.e1e1213.e2
play a major role in the pathogenesis of the disease. Previous data (Ismail et al., 2012) demonstrated the presence of the mutation in patients with MS and ALS, but not MS alone. Taken together, these findings suggest that the expansion could be causative of ALS independent of the co-occurrence of MS.
Acknowledgements This work was supported by grant Italian Ministry of Health (Ricerca Corrente) and Fondazione Monzino. Appendix A. Supplementary data
Disclosure statement The authors have no actual or potential conflicts of interest.
Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j.neurobiolaging. 2013.10.096.
