Three-dimensional tissue engineered skeletal muscle modelling facioscapulohumeral muscular dystrophy

Brain, vol. 148, pp. 1723–1739

Abstract

Facioscapulohumeral muscular dystrophy (FSHD) is caused by sporadic misexpression of the transcription factor double homeobox 4 (DUX4) in skeletal muscles. So far, monolayer cultures and animal models have been used to study the disease mechanism of FSHD and for development of FSHD therapy, but these models do not fully recapitulate the disease and there is a lack of knowledge on how DUX4 misexpression leads to skeletal muscle dysfunction. To overcome these barriers, we have developed a 3D tissue engineered skeletal muscle (3D-TESM) model by generating genetically matched myogenic progenitors from human induced pluripotent stem cells of three mosaic FSHD patients. 3D-TESMs derived from genetically affected myogenic progenitors recapitulated pathological features including DUX4 and DUX4 target gene expression, smaller myofibre diameters and reduced absolute forces upon electrical stimulation. RNA-sequencing data illustrated increased expression of DUX4 target genes in 3D-TESMs compared with 2D myotubes, and cellular differentiation was improved by 3D culture conditions. Treatment of 3D-TESMs with three different small molecules identified in drug development screens in 2D muscle cultures showed no improvements, and sometimes even declines, in contractile force and sarcomere organization. These results suggest that these compounds either have a detrimental effect on the formation of 3D-TESMs, an effect that might have been overlooked or was challenging to detect in 2D cultures and in vivo models, and/or that further development of the 3D-TESM model is needed. In conclusion, we have developed a 3D skeletal muscle model for FSHD that can be used for preclinical research focusing on DUX4 expression and downstream pathways of FSHD in relationship to contractile properties. In the future, we expect that this model can also be used for preclinical drug screening.

Authors 17

1. Marnix Franken Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

2. Erik van der Wal Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

3. Dongxu Zheng Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

4. Bianca den Hamer Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

5. Patrick J. van der Vliet Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

6. Richard J.L.F. Lemmers Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

7. Anita van den Heuvel Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

8. Alexandra L Dorn Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

9. Cas G A Duivenvoorden Aachen Department of Human Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

   Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

10. Stijn L.M. in ‘t Groen

    Erasmus MC · Erasmus University Rotterdam

    Affiliation as printed

    Center for Lysosomal and Metabolic Diseases, Erasmus University Medical Center , 3015 GE Rotterdam ,

    Department of Clinical Genetics, Erasmus University Medical Center , 3015 GD, Rotterdam ,

    Department of Pediatrics, Erasmus University Medical Center , 3015 GE Rotterdam ,

    Department of Clinical Genetics, Erasmus Medical Center, 3015 GD, Rotterdam, The Netherlands

11. Christian M.A.H. Freund Aachen Leiden hiPSC Centre Department of Anatomy and Embryology

    Leiden University Medical Center

    Affiliation as printed

    Leiden hiPSC Centre, Department of Anatomy and Embryology, Leiden University Medical Center , 2333 ZA, Leiden ,

    Leiden hiPSC Centre, Department of Anatomy and Embryology, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

12. Bert Eussen

    Erasmus MC · Erasmus University Rotterdam

    Affiliation as printed

    Department of Clinical Genetics, Erasmus University Medical Center , 3015 GD, Rotterdam ,

    Department of Clinical Genetics, Erasmus Medical Center, 3015 GD, Rotterdam, The Netherlands

13. Rabi Tawil

    University of Rochester Medicine

    Affiliation as printed

    Department of Neurology, University of Rochester Medical Center , Rochester, NY 14642 ,

    Department of Neurology, University of Rochester Medical Center, Rochester, NY 14642, USA

14. Baziel G.M. van Engelen

    Radboud University Nijmegen · Radboud University Medical Center · Donders Centre for Neuroscience

    Affiliation as printed

    Department of Neurology, Donders Centre of Neuroscience, Radboud University Medical Centre , 6525 GA, Nijmegen ,

    Department of Neurology, Donders Centre of Neuroscience, Radboud University medical Centre, 6525 GA, Nijmegen, The Netherlands

15. W.W.M. Pim Pijnappel

    Erasmus MC · Erasmus University Rotterdam

    Affiliation as printed

    Center for Lysosomal and Metabolic Diseases, Erasmus University Medical Center , 3015 GE Rotterdam ,

    Department of Clinical Genetics, Erasmus University Medical Center , 3015 GD, Rotterdam ,

    Department of Pediatrics, Erasmus University Medical Center , 3015 GE Rotterdam ,

    Department of Clinical Genetics, Erasmus Medical Center, 3015 GD, Rotterdam, The Netherlands

16. Silvère M. van der Maarel Aachen Department of Human Genetics

    Leiden University Medical Center

    Affiliation as printed

    Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

    Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands

17. Jessica C. de Greef corresponding Aachen Department of Human Genetics

    Leiden University Medical Center

    Affiliation as printed

    Department of Human Genetics, Leiden University Medical Center , 2333 ZA, Leiden ,

    Department of Human Genetics, Leiden University Medical Center, 2333 ZA, Leiden, The Netherlands