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Calpain 3 deficiency is associated with myonuclear apoptosis and profound perturbation of the IκBα/NF-κB pathway in limb-girdle muscular dystrophy type 2A

Nature Medicine volume 5, pages 503–511 (1999)Cite this article

An Erratum to this article was published on 01 July 1999

Limb-girdle muscular dystrophy type 2A results from calpain 3 deficiency. In deltoid muscle biopsies of limb-girdle muscular dystrophy type 2A, myonuclear apoptosis was correlated here with altered subcellular distribution of IκBα and NF-κB, resulting in sarcoplasmic sequestration of NF-κB. Calpain 3-dependent IκBα degradation was shown to take place in a co-expression cell system. IκBα overexpression prevented nuclear accumulation of NF-κB and led to apoptosis in primary cultures of human myogenic satellite cells. The data indicate that calpain 3, present in healthy muscle as sarcoplasmic and nuclear forms, controls IκBα turnover and indirectly regulates NF-κB-dependent expression of survival genes.

Limb girdle muscular dystrophies (LGMDs) are characterized by progressive symmetrical atrophy and weakness of the proximal limb, scapular pelvic girdle and trunk muscles, without affecting facial muscles1. The diagnosis for autosomal recessive LGMDs remained, until recently, one of the most challenging and difficult to establish2,3. The identification of the skeletal muscle-specific CAPN3 gene, which encodes calpain 3, a calcium-activated cystein protease, now permits genetic or molecular diagnosis4,5,6. This is the first case, to our knowledge, of an autosomal muscular dystrophy caused by a defect in a gene encoding a protein with enzymatic rather than structural properties. The substrate(s) of calpain 3 and the pathways involved in the development of LGMD2A are still unknown. Calpain 3 mRNA in skeletal muscle is 1,000% more abundant than mRNAs of ubiquitous calpains 1 and 2 (ref. 7).

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Figure 1: Calpain 3 localizes in myofibrils and myonuclei in healthy skeletal muscle.
Figure 2: Apoptosis in LGMD2A deltoid muscular biopsies.
Figure 3: TUNEL-positive nuclei coexist with TUNEL-negative nuclei within the same myofiber.
Figure 4: LGMD2A TUNEL-positive nuclei are IκBα-positive and p65NF-κB-negative.
Figure 5: p65NF-κB accumulates in LGMD2A patients.
Figure 6: IκBα is degraded in Sf9 cells that express a recombinant form of calpain 3.
Figure 7: Transient overexpression of IκBα prevents NF-κB nuclear accumulation and induces apoptosis in primary culture of human muscle satellite cells.

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Acknowledgements

We are particularly indebted to the L5988 and L7111 families members and A21 and A13 for their continued and invaluable cooperation. We also thank P. Farah, A. Megarbane, J. Ducret, H. Aydenian, J. Maarrawi, G. Abadjian, N. Salem and the clinicians of the North Lebanon. We also acknowledge J. Demaille, I. Touitou, B. Vallet, N. Romero, G. Restagno, A.M. Cobo, I. Penisson, A. Vila and X. Ferrer as well as A. Sahuquet, H. Lelouard and S. Bonavaud. We are also indebted to J.L. Vayssière and B. Mignotte for discussions and S. Artero for statistical analysis. This work was supported by grants from the Saint-Joseph and Montpellier II Universities, the Lebanese and French CNRS (International Programs of Scientific Cooperation), the Association Française contre les Myopathies (AFM), the INSERM and the Ambassade de France.

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Authors and Affiliations

  1. Laboratoire de Dynamique Moléculaire des Interactions Membranaires, CNRS-UMR 5539, et Laboratoire de Motilité Cellulaire (EPHE), Université Montpellier II, case 107, Place Eugène Bataillon, Montpellier, 34095 cedex 5, France

    Stephen Baghdiguian, Marianne Martin, Catherine Astier & Paul Mangeat

  2. Généthon and URA CNRS 1992, 1 rue de l'Internationale, BP 60, Evry, 91002, France

    Isabelle Richard, Nathalie Bourg & Jacques S. Beckmann

  3. INSERM-U 300, Faculté de Pharmacie, Av. Ch. Flahault, 34060 cedex 1, Montpellier, France

    Françoise Pons

  4. School of Biological and Medical Sciences, University of St. Andrews, Fife, KY16 9AL, Scotland

    Ronald T. Hay

  5. Hôtel-Dieu et Faculté de Médecine, Université, Saint Joseph, Beyrouth, Liban

    Raymond Chemaly, Georges Halaby & Jacques Loiselet

  6. Neurobiology Department, University Medical School Framlington place, NewCastle Upon Tyne, NE2 4HH, UK

    Louise V. B. Anderson

  7. Hospital Ntra SRA., Aranzazu, Dept. of Neurology, Apdo correos 477, San Sebastian, 20080, Spain

    Adolfo Lopez de Munain

  8. INSERM-U153/CNRS-UA614, 17 rue du Fer à moulin, Paris, 75005, France

    Michel Fardeau

  9. Laboratoire d'ImmunoGénétique Moléculaire, Institut de Génétique Humaine CNRS UPR 1142 and Université Montpellier II, Place Eugène Bataillon, 34095 5, Montpellier cedex, France

    Gérard Lefranc

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Baghdiguian, S., Martin, M., Richard, I. et al. Calpain 3 deficiency is associated with myonuclear apoptosis and profound perturbation of the IκBα/NF-κB pathway in limb-girdle muscular dystrophy type 2A. Nat Med 5, 503–511 (1999). https://doi.org/10.1038/8385

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