Biomechanical characterization of a desminopathy in primary human myoblasts

Bonakdar, Navid; Łuczak, Justyna; Lautscham, Lena; Czonstke, Maja; Koch, Thorsten; Mainka, Astrid; Jungbauer, Tajana; Goldmann, Wolfgang H.; Schröder, Rolf; Fabry, Ben

doi:10.1016/j.bbrc.2012.02.083

Cited by 43 publications

(43 citation statements)

References 15 publications

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“…These studies demonstrated an increased stiffness in heterozygous myoblasts, indicating a reduced deformability compared to immortalized wt myoblasts. The latter findings mirror previous results in cultured primary myoblasts from a patient with a het R350P desmin mutation 27 . The analysis of hom immortalized R349P desmin-knock-in myoblasts, however, revealed lateral compliance levels that were in the same range as in wt cells.…”

Section: Discussionsupporting

confidence: 91%

“…The bead displacement (d) after a step increase of the force (F) followed a power law with time (t) as described by ref. 27. The cell’s lateral compliance J 0 , which is inversely proportional to its stiffness, was determined from the creep response J ( t ) of the cells by fitting the displacement with the typical power law response , where .…”

Section: Resultsmentioning

confidence: 99%

“…For compliance recordings of immortalized wt, het, and hom R349P cultured myoblasts, a magnetic tweezer device was used as described previously 27 . Details are given in the SI.…”

Section: Methodsmentioning

confidence: 99%

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Early signs of architectural and biomechanical failure in isolated myofibers and immortalized myoblasts from desmin-mutant knock-in mice

Diermeier

Iberl

Vetter

et al. 2017

Sci Rep

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In striated muscle, desmin intermediate filaments interlink the contractile myofibrillar apparatus with mitochondria, nuclei, and the sarcolemma. The desmin network’s pivotal role in myocytes is evident since mutations in the human desmin gene cause severe myopathies and cardiomyopathies. Here, we investigated skeletal muscle pathology in myofibers and myofibrils isolated from young hetero- and homozygous R349P desmin knock-in mice, which carry the orthologue of the most frequent human desmin missense mutation R350P. We demonstrate that mutant desmin alters myofibrillar cytoarchitecture, markedly disrupts the lateral sarcomere lattice and distorts myofibrillar angular axial orientation. Biomechanical assessment revealed a high predisposition to stretch-induced damage in fiber bundles of R349P mice. Notably, Ca2 +-sensitivity and passive myofibrillar tension were decreased in heterozygous fiber bundles, but increased in homozygous fiber bundles compared to wildtype mice. In a parallel approach, we generated and subsequently subjected immortalized heterozygous R349P desmin knock-in myoblasts to magnetic tweezer experiments that revealed a significantly increased sarcolemmal lateral stiffness. Our data suggest that mutated desmin already markedly impedes myocyte structure and function at pre-symptomatic stages of myofibrillar myopathies.

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Section: Discussionsupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

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Early signs of architectural and biomechanical failure in isolated myofibers and immortalized myoblasts from desmin-mutant knock-in mice

Diermeier

Iberl

Vetter

et al. 2017

Sci Rep

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“…Finally, to assess whether 4-PBA could improve the resilience of Plec -/-myotubes against mechanical strain, we exposed cells spread on flexible membranes to cyclic stretch and assessed their vulnerability by microscopically monitoring their substrate detachment rates (17,18). In this experiment, 51% of differentiated Plec -/-myotubes detached, compared with only 15% of their Plec +/+ counterparts.…”

Section: Phenotype Amelioration Through Chemical Chaperone 4-pba Treamentioning

confidence: 99%

Chemical chaperone ameliorates pathological protein aggregation in plectin-deficient muscle

Winter¹,

Staszewska²,

Mihailovska³

et al. 2014

J. Clin. Invest.

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The ubiquitously expressed multifunctional cytolinker protein plectin is essential for muscle fiber integrity and myofiber cytoarchitecture. Patients suffering from plectinopathy-associated epidermolysis bullosa simplex with muscular dystrophy (EBS-MD) and mice lacking plectin in skeletal muscle display pathological desmin-positive protein aggregation and misalignment of Z-disks, which are hallmarks of myofibrillar myopathies (MFMs). Here, we developed immortalized murine myoblast cell lines to examine the pathogenesis of plectinopathies at the molecular and single cell level. Plectin-deficient myotubes, derived from myoblasts, were fully functional and mirrored the pathological features of EBS-MD myofibers, including the presence of desmin-positive protein aggregates and a concurrent disarrangement of the myofibrillar apparatus. Using this cell model, we demonstrated that plectin deficiency leads to increased intermediate filament network and sarcomere dynamics, marked upregulation of HSPs, and reduced myotube resilience following mechanical stretch. Currently, no specific therapy or treatment is available to improve plectin-related or other forms of MFMs; therefore, we assessed the therapeutic potential of chemical chaperones to relieve plectinopathies. Treatment with 4-phenylbutyrate resulted in remarkable amelioration of the pathological phenotypes in plectin-deficient myotubes as well as in plectin-deficient mice. Together, these data demonstrate the biological relevance of the MFM cell model and suggest that this model has potential use for the development of therapeutic approaches for EBS-MD.

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“…Abnormal desmin filament elasticity has been postulated as a contributory mechanism of disease progression. A recent study performed in primary cultured myoblasts obtained from a patient revealed abnormal mechanical properties of affected muscle cells, which acquire increased stiffness and higher vulnerability to mechanical stretch [45■]. …”

Section: Progress In Studies Of the Previously Identified Myofibrillamentioning

confidence: 99%

Myofibrillar myopathies

Olivé

Kley

Goldfarb

2013

Current Opinion in Neurology

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Purpose of review Myofibrillar myopathies (MFMs) are a heterogeneous group of skeletal and cardiac muscle diseases. In this review, we highlight recent discoveries of new genes and disease mechanisms involved in this group of disorders. Recent findings The advent of next-generation sequencing technology, laser microdissection and mass spectrometry-based proteomics has facilitated the discovery of new MFM causative genes and pathomechanisms. New mutations have also been discovered in ‘older’ genes, helping to find a classification niche for MFM-linked disorders showing variant phenotypes. Cell transfection experiments using primary cultured myoblasts and newer animal models provide insights into the pathogenesis of MFMs. Summary An increasing number of genes are involved in the causation of variant subtypes of MFM. The application of modern technologies in combination with classical histopathological and ultrastructural studies is helping to establish the molecular diagnosis and reach a better understanding of the pathogenic mechanisms of each MFM subtype, thus putting an emphasis on the development of specific means for prevention and therapy of these incapacitating and frequently fatal diseases.

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Biomechanical characterization of a desminopathy in primary human myoblasts

Cited by 43 publications

References 15 publications

Early signs of architectural and biomechanical failure in isolated myofibers and immortalized myoblasts from desmin-mutant knock-in mice

Early signs of architectural and biomechanical failure in isolated myofibers and immortalized myoblasts from desmin-mutant knock-in mice

Chemical chaperone ameliorates pathological protein aggregation in plectin-deficient muscle

Myofibrillar myopathies

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