title: Impaired nuclear functions lead to increased senescence and inefficient differentiation in human myoblasts with a dominant p.R545C mutation in the LMNA gene.
abstract: We have studied myoblasts from a patient with a severe autosomal dominant Emery-Dreifuss muscular dystrophy (AD-EDMD) caused by an arginine 545 to cystein point mutation (p.R545C) in the carboxy-terminal domain of the lamin A/C gene. This mutation has pleiotropic cellular effects on these myoblasts as demonstrated by nuclear structural defects, exhibiting lobulations which increase with cell passages in culture. The organization of both lamin A/C and its inner nuclear membrane partner emerin are altered, eventually showing a honeycomb pattern upon immunofluorescence microscopy. In addition, the distribution of histone H3 trimethylated at lysine 27 and of phosphorylated RNA polymerase II, markers of inactive and active chromatin domains, respectively, are altered suggesting an impact on gene expression. Patient myoblasts also presented a high index of senescence in ex vivo culture. Moreover, our data show for the first time in an AD-EDMD context that the 20S core particle of the proteasome was inactivated. With cell passages, the 20S core protein progressively accumulated into discrete nuclear foci that largely colocalized with promyelocytic leukemia (PML) bodies while p21 accumulated throughout the nuclear compartment. Proteasome inactivation has been linked to normal cellular ageing. Our data indicate that it may also contribute to premature senescence in AD-EDMD patient myoblasts. Finally, when transferred to low-serum medium, patient myoblasts were deficient in ex vivo differentiation, as assessed by the absence of myotube formation and myogenin induction. Altogether, these data suggest that the LMNA mutation p.R545C impairs both proliferation and differentiation capacities of myoblasts as part of the pathogenesis of AD-EDMD.
authors: Division of Electron Microscopy, Biocenter, University of Würzburg, Am Hubland, D-97074 Würzburg, Germany.
authors: Sebastian
authors: S
authors: Kandert
authors: Manfred
authors: M
authors: Wehnert
authors: Clemens R
authors: CR
authors: Müller
authors: Brigitte
authors: B
authors: Buendia
authors: Marie-Christine
authors: MC
authors: Dabauvalle
mesh_terms: D000375:Aging / Q000235:genetics*; D019943:Amino Acid Substitution; D000906:Antibodies / Q000378:metabolism; D006690:Bisbenzimidazole / Q000378:metabolism; D002232:Carbocyanines / Q000378:metabolism; D016022:Case-Control Studies; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics*; D002467:Cell Nucleus / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D002478:Cells, Cultured; D003545:Cysteine / Q000378:metabolism; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D034904:Lamin Type A / Q000235:genetics*; D008297:Male; D020389:Muscular Dystrophy, Emery-Dreifuss / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D020125:Mutation, Missense; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D017354:Point Mutation; D014966:Xanthenes / Q000378:metabolism
chemical_list: D000906:Antibodies; D002232:Carbocyanines; D005456:Fluorescent Dyes; D034904:Lamin Type A; D014966:Xanthenes; C089187:cyanine dye 3; C034657:Texas red; D016650:Fluorescein-5-isothiocyanate; D003545:Cysteine; D006690:Bisbenzimidazole