A Human Pluripotent Stem Cell Model of Facioscapulohumeral Muscular Dystrophy-Affected Skeletal Muscles

Abstract

Facioscapulohumeral muscular dystrophy (FSHD) represents a major unmet clinical need arising from the progressive weakness and atrophy of skeletal muscles. The dearth of adequate experimental models has severely hampered our understanding of the disease. To date, no treatment is available for FSHD. Human embryonic stem cells (hESCs) potentially represent a renewable source of skeletal muscle cells (SkMCs) and provide an alternative to invasive patient biopsies. We developed a scalable monolayer system to differentiate hESCs into mature SkMCs within 26 days, without cell sorting or genetic manipulation. Here we show that SkMCs derived from FSHD1-affected hESC lines exclusively express the FSHD pathogenic marker double homeobox 4 and exhibit some of the defects reported in FSHD. FSHD1 myotubes are thinner when compared with unaffected and Becker muscular dystrophy myotubes, and differentially regulate genes involved in cell cycle control, oxidative stress response, and cell adhesion. This cellular model will be a powerful tool for studying FSHD and will ultimately assist in the development of effective treatments for muscular dystrophies. STEM CELLS TRANSLATIONAL MEDICINE 2016;5:1145-1161 SIGNIFICANCEThis work describes an efficient and highly scalable monolayer system to differentiate human pluripotent stem cells (hPSCs) into skeletal muscle cells (SkMCs) and demonstrates disease-specific phenotypes in SkMCs derived from both embryonic and induced hPSCs affected with facioscapulohumeral muscular dystrophy. This study represents the first human stem cell-based cellular model for a muscular dystrophy that is suitable for high-throughput screening and drug development.

GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells

“…These cells were generated using a modified protocol that used DAPT instead of FGF2 followed by FACS with NCAM + /HNK − markers for purification. Caron et al 36 . pre-plated cells 10 days after starting the differentiation protocol.…”

A Myogenic Double-Reporter Human Pluripotent Stem Cell Line Allows Prospective Isolation of Skeletal Muscle Progenitors

“…Later on, transgene-free approaches were developed using directed differentiation toward mesodermal-paraxial lineages. These efforts employed by tuning major signaling pathways governing stem cell differentiation toward mesoderm and muscle, such as WNT, BMP, and transforming growth factor β (TGF-β) (Barberi et al, 2007; Borchin et al, 2013; Caron et al, 2016; Chal et al, 2015; Hicks et al, 2018; Shelton et al, 2014; Xi et al, 2017; Xu et al, 2013). Although these methods were successful in inducing skeletal myogenesis, parallel generation of unwanted cell types hindered the purity of the myogenic cells, as recently reported by our collaborators (Kim et al, 2017).…”

Differentiation and sarcomere formation in skeletal myocytes directly prepared from human induced pluripotent stem cells using a sphere-based culture

“…While many protocols require cell sorting and/or rely on exogenous expression of myogenic genes such as PAX3, PAX7, and MYOD (Abujarour et al, 2014; Darabi et al, 2012; Maffioletti et al, 2015; Skoglund et al, 2014; Tanaka et al, 2013), more recent advances have been made with the application of small molecules and growth factors to directly promote myogenic differentiation from human iPSCs (Barberi et al, 2007; Borchin et al, 2013; Caron et al, 2016; Chal et al, 2016; Chal et al, 2015; Choi et al, 2016; Hosoyama et al, 2014; Hwang et al, 2013; Shelton et al, 2014; Xu et al, 2013). Our group recently reported a unique method for the derivation of myogenic progenitors from human pluripotent cells using free-floating spherical culture (Hosoyama et al, 2014).…”