Heidi Zhuge (Bochum / DE), Dr Britta Eggers (Bochum / DE), Nassam Daya (Bochum / DE), Dr. med. Anne-Katrin Güttsches (Bochum / DE), Dr. Andreas Unger (Münster / DE), Dr. Peter van der Ven (Bonn / DE), Dr Dieter Fürst (Bonn / DE), Dr Anne Bigot (Paris / FR), Dr Vincent Mouly (Paris / FR), Prof Dr Katrin Marcus (Bochum / DE), PD Dr.rer nat Andreas Roos (Bochum / DE), Prof. Dr. Matthias Vorgerd (Bochum / DE)
Abstract-Text (inkl. Referenzen)
Background: The p.(W2710X)_FLNc nonsense variant was the first dominant mutation linked to filaminopathies as a subgroup of myofibrillar myopathies. Filaminopathies are associated with muscle weakness, sarcomere lesions and protein aggregation from the Z-disk (varying depending on the mutation). As the pathophysiology is still incompletely understood and no treatment is in place, establishment of an in vitro human model is crucial.
Methods: Differentiation of p.(W2710X)_FLNc and wildtype myoblasts to striated myotubes was achieved by applying electrical pulse stimulation (EPS). Immunological, ultrastructural and proteomic studies were performed in addition to proteasome - and MTT-assays towards phenotyping of the in vitro model. Treatment with Arimoclomol was tested to observe the effect on the defined phenotype.
Results: Mutant myotubes showed more lesions similar to early-stage pathology in patients compared to control cells. Proteomic data revealed dysregulation of a variety of proteins also affecting metabolic pathways accompanied by altered proteasome function and viability in mutant cells. Arimoclomol was able to significantly improve p.(W2710X) cell vitality and reduce cell toxicity.
Conclusion: Due to a robust differentiation protocol, the immortalized p.(W2710X) cell model constitutes a reliable and valid cell model for early-stage pathogenesis and therapeutic targets without the limitation of having to rely on patient biopsy material.