Novel Missense CAPN3 Mutation Responsible for Adult-Onset Limb Girdle Muscular Dystrophy with Calves Hypertrophy
- 10 Downloads
CAPN3 gene encodes for calpain-3; this protein is a calcium-dependent intracellular protease. Deficiency of this enzyme leads to weakness of the proximal limb muscles and pelvic and shoulder girdles, the so-called limb-girdle muscular dystrophy type 2A (LGMD2A). Here, we reported the case of a Tunisian patient with LGMD2A associated with a novel missense mutation (c.T1681C/p.Y561H). A 61-year-old man, with consanguineous parents, was referred for gait difficulties and slowly progressive proximal weakness of the four limbs associated with moderate hypertrophy of the calves but his facial muscles were unaffected. Electromyography showed that the profile was myopathic pattern and creatine kinase (CK) level was high. Muscle biopsy processing included routine histological, immunohistochemical, and Western Blot reactions, using a panel of antibodies directed against dystrophin, dysferlin, calpain-3, sarcoglycan α, β, γ, and δ. For mutation analysis, we designed an NGS-based screening. Immunological analyses demonstrated a total deficiency in calpain-3 and δ-sarcoglycan, and a reduced expression of dysferlin. The genetic study yielded a homozygous missense mutation (c.T1681C) of the 13th exon of the CAPN3 gene. The mutation found in our patient (c.T1681C/p.Y561H) has not been previously reported. It is responsible for complete calpain-3 and δ-sarcoglycan deficiency and reduced dysferlin expression. The genetic study is mandatory in such cases with multiple-protein deficiency and ambiguous results of immune-histology and Western Blot studies.
KeywordsMuscular dystrophies Immunohistochemistry Western Blot CAPN3 protein DNA sequencing LGMD2A
We would like to thank the genomic platform members in IMRB (Institut Mondor de Recherche Biomédicale), all the staff of the Department of Neurology, as well as all the staff of the Laboratory of Pathology, Habib Bourguiba University Hospital in Sfax. We also thank Chokri Khalaf for proofreading the study.
This work was supported by the “Association Française contre les Myopathies” Translamuscle #19507.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- L Brenguier, I Richard, P Dinçer, C Roudaut, B Bady, J M Burgunder, R Chemaly, C A Garcia, G Halaby, C E Jackson, D M Kurnit, G Lefranc, C Legum, J Loiselet, L Merlini, A Nivelon-Chevallier, E Ollagnon-Roman, G Restagno, H Topaloglu, and J S Beckmann, (1997) Multiple independent molecular etiology for limb-girdle muscular dystrophy type 2A patients from various geographical origins. Am J Hum Genet. 60(5): 1128–1138.Google Scholar
- Chae J1, Minami N, Jin Y, Nakagawa M, Murayama K, Igarashi F, Nonaka I. (2001). Calpain 3 gene mutations: genetic and clinico-pathologic findings in limbgirdle muscular dystrophy. Neuromuscul Disord (6-7):547-55.Google Scholar
- Chrobáková T, Hermanová M, Kroupová I, Vondráček P, Mařı́ková T', Mazanec R, Zámečnı́k J, Staněk J, Havlová M, Fajkusová L (2004) Mutations in Czech LGMD2A patients revealed by analysis of calpain3 mRNA and their phenotypic outcome. Neuromuscul Disord 14:659–665. https://doi.org/10.1016/j.nmd.2004.05.005 CrossRefGoogle Scholar
- Dinçer P1, Leturcq F, Richard I, Piccolo F, Yalnizoglu D, de Toma C, Akçören Z, Broux O, Deburgrave N, Brenguier L, Roudaut C, Urtizberea JA, Jung D, Tan E, Jeanpierre M, Campbell KP, Kaplan JC, Beckmann JS, Topaloglu H. (1997). A biochemical, genetic, and clinical survey of autosomal recessive limb girdle muscular dystrophies in Turkey. Ann Neurol 42(2):222-9.Google Scholar
- Fadaee M, Kariminejad A, Fattahi Z, Nafissi S, Godarzi HR, Beheshtian M, Vazehan R, Akbari MR, Kahrizi K, Najmabadi H (2016) Report of limb-girdle muscular dystrophy type 2a in 6 Iranian patients, one with a novel deletion in CAPN3 gene. Neuromuscul Disord 26:277–282. https://doi.org/10.1016/j.nmd.2016.02.003 CrossRefGoogle Scholar
- Fanin M, Fulizio L, Nascimbeni AC, Spinazzi M, Piluso G, Ventriglia VM, Ruzza G, Siciliano G, Trevisan CP, Politano L, Nigro V, Angelini C (2004) Molecular diagnosis in LGMD2A: Mutation analysis or protein testing?. Human Mutation 24 (1):52-62.Google Scholar
- Fanin M, Nascimbeni AC, Angelini C (2006) Screening of calpain-3 autolytic activity in LGMD muscle: a functional map of CAPN3 gene mutations. Journal of Medical Genetics 44 (1):38-43.Google Scholar
- Fanin M, Nascimbeni AC, Tasca E, Angelini C (2009) How to tackle the diagnosis of limb-girdle muscular dystrophy 2A. European Journal of Human Genetics 17 (5):598-603.Google Scholar
- Hadj Salem I, Kamoun F, Louhichi N, Rouis S, Mziou M, Fendri‑Kriaa N, Makni‑Ayadi F, Triki C, Fakhfakh F (2011) Mutations in LAMA2 and CAPN3 genes associated with genetic and phenotypic heterogeneities within a single consanguineous family involving both congenital and progressive muscular dystrophies. Biosci Rep 31:125–135. https://doi.org/10.1042/BSR20100026 CrossRefGoogle Scholar
- Huang Y, Verheesen P, Roussis A, Frankhuizen W, Ginjaar I, Haldane F, Laval S, Anderson LVB, Verrips T, Frants RR, de Haard H, Bushby K, den Dunnen J, van der Maarel SM (2005) Protein studies in dysferlinopathy patients using llama-derived antibody fragments selected by phage display. Eur J Hum Genet 13:721–730. https://doi.org/10.1038/sj.ejhg.5201414 CrossRefGoogle Scholar
- I. Richard, C. Roudaut, A. Saenz, R. Pogue, J.E.M.A. Grimbergen, L.V.B. Anderson, C. Beley, A-M Cobo, C. de Diego, B. Eymard, P. Gallano, H.B. Ginjaar, A. Lasa, C. Pollitt, H. Topaloglu, J.A. Urtizberea, M. de Visser, A. van der Kooi, K. Bushby, E. Bakker, A. Lopez de Munain, M. Fardeau, J.S. Beckmann, (1999) Calpainopathy—A Survey of Mutations and Polymorphisms. The American Journal of Human Genetics 64 (6):1524-1540.Google Scholar
- Jackman RW et al (2004) The molecular basis of skeletal muscle atrophy. Am J Physiol Cell Physiol 4:834–4–843.Google Scholar
- Jennifer M. Thompson M, Moore SA, Liewluck T (2018) A novel CAPN3 mutation in late-onset limb-girdle muscular dystrophy with early respiratory insufficiency. Journal of Clinical Neuroscience 53:229-231.Google Scholar
- Nigro V, Savarese M (2014) Genetic basis of limb-girdle muscular dystrophies: the 2014 update. Acta Myol 33:1–12.Google Scholar
- Ono Y, Shimada H, Sorimachi H, Richard I, Saido TC, Beckmann JS, Ishiura S, Suzuki K (1998) Functional defects of a muscle-specific Calpain, p94, caused by mutations associated with limb-girdle muscular dystrophy type 2A. J Biol Chem 273:17073–17078. https://doi.org/10.1074/jbc.273.27.17073 CrossRefGoogle Scholar
- Piluso G, Politano L, Aurino S, Fanin M, Ricci E, Ventriglia VM, Belsito A, Totaro A, Saccone V, Topaloglu H, Nascimbeni AC, Fulizio L, Broccolini A, Canki-Klain N, Comi LI, Nigro G, Angelini C, Nigro V (2005) Extensive scanning of the calpain-3 gene broadens the spectrum of LGMD2A phenotypes. J Med Genet 42:686–693. https://doi.org/10.1136/jmg.2004.028738 CrossRefGoogle Scholar
- Rumiko Izumi, Tetsuya Niihori, Toshiaki Takahashi, Naoki Suzuki, Maki Tateyama, Chigusa Watanabe, Kazuma Sugie, Hirotaka Nakanishi, Gen Sobue, Masaaki Kato, Hitoshi Warita, Yoko Aoki, Masashi Aoki, (2015) Genetic profile for suspected dysferlinopathy identified by targeted next-generation sequencing. Neurology Genetics 1 (4):e36.Google Scholar
- Srujana et al (2012) Quality control in aminoacyl-tRNA synthesis. Advances in Protein Chemistry and Structural Biology. Adv Protein Chem Struct Bio:1–43. https://doi.org/10.1016/b978-0-12-386497-0.00001-3
- Straub V, Murphy A, Udd B, LGMD workshop study group (2018) 229th ENMC international workshop: limb-girdle muscular dystrophies - nomenclature and reformed classification Naarden, the Netherlands, 17-19 March 2017. Neuromuscul Disord 28:702–710. https://doi.org/10.1016/j.nmd.2018.05.007 CrossRefGoogle Scholar