Patient-specific iPSC-derived 2D myotubes and 3D musculoids harboring FLNC variants exhibited poor sarcomeric organization, protein aggregation, and ultrastructural abnormalities compared to controls.
The developed iPSC-derived 2D and 3D skeletal muscle models successfully recapitulate the pathological hallmarks of filaminopathies, providing a platform for studying disease mechanisms and potential therapeutic interventions.
Abstract Background Filaminopathies, caused by pathogenic FLNC variants, are rare neuromuscular disorders characterized by protein aggregation, z-disk pathology and lead to progressive muscle weakness and/or cardiomyopathies. Methods To address the lack of existing filaminopathy models in skeletal muscle, we developed a patient-specific cellular platform using induced pluripotent stem cells (iPSCs) harboring two truncating filamin C (FLNc) variants (p.Q1662X, p.Y2704X). Employing a developmental human skeletal muscle organoid hSMO model, we enrich for myogenic progenitor cells that are further differentiated into functional myotubes through 2D and 3D approaches (myotubes and musculoids). Results The 2D myotubes exhibited poor sarcomeric organization and hallmarks of filaminopathies, including protein aggregation and proteostatic dysfunction, marked by elevated aggresome formation and an increased basal autophagic flux. The 3D musculoids revealed ultrastructural abnormalities and enabled the identification of novel disease-associated proteins involved in ER stress and protein folding (e.g. DNAJC10) through proteomic analysis. Proteomic findings were additionally validated in 2D cultures and in corresponding patient-derived muscle biopsies enhancing the model’s translational value. Conclusions Our model is suitable to monitor aspects of filaminopathies’ pathogenesis and to investigate possible therapeutic interventions with quantitative readouts.
Daya et al. (Thu,) conducted a other in Filaminopathies (n=3). iPSC-derived 2D myotubes and 3D musculoids vs. Unaffected control iPSCs was evaluated. Patient-specific iPSC-derived 2D myotubes and 3D musculoids harboring FLNC variants exhibited poor sarcomeric organization, protein aggregation, and ultrastructural abnormalities compared to controls.