Identification of Terminal Differentiation Enhancers in Human Embryonic Stem Cell Derived Skeletal Muscle Cells

Abstract

Skeletal myogenesis is a tightly coordinated process resulting from the temporal expression of signalling cascades that specify myogenic cell fate. Identification of signalling pathways and small molecules that can modulate this developmental process, continues to be an active area of research. Utilising the pluripotent nature of human embryonic stem cells (hESC) and combined with next generation sequencing, we demonstrate our in vitro skeletal muscle differentiation system accurately recapitulate major skeletal muscle developmental process. We show myotubes formation can be further enhanced using a combination of anabolic factors and myokines. Multi-omics analysis revealed oxidative phosphorylation as the major up-regulated pathway, suggesting energy metabolism is coupled to enhanced skeletal muscle differentiation. Finally, to identify novel drug candidates that could reinforce muscle strength, we performed a high throughput drug screening of over 1000 drugs in hESC derived skeletal muscle cells (hESC-SkMC) and identified several candidate compounds that significantly increased the muscle marker Myosin Heavy Chain (MyHC) expression level. We further demonstrate this enhanced muscle differentiation is also closely associated with an up-regulation of energy metabolism. Together, this work presents a genetic dissection of hESC-SkMC development in vitro, which may assist in the identification and development of more effective drug therapies targeting skeletal muscle development or diseases.