MicroRNA control of the myogenic cell transcriptome and proteome: the role of miR-16

MicroRNAs (miRs) control stem cell biology and fate. Ubiquitously expressed and conserved miR-16 was the first miR implicated in tumorigenesis. miR-16 is low in muscle during developmental hypertrophy and regeneration. It is enriched in proliferating myogenic progenitor cells but is repressed during differentiation. The induction of miR-16 blocks myoblast differentiation and myotube formation, whereas knockdown enhances these processes. Despite a central role for miR-16 in myogenic cell biology, how it mediates its potent effects is incompletely defined. In this investigation, global transcriptomic and proteomic analyses after miR-16 knockdown in proliferating C2C12 myoblasts revealed how miR-16 influences myogenic cell fate. Eighteen hours after miR-16 inhibition, ribosomal protein gene expression levels were higher relative to control myoblasts and p53 pathway-related gene abundance was lower. At the protein level at this same time point, miR-16 knockdown globally upregulated tricarboxylic acid (TCA) cycle proteins while downregulating RNA metabolism-related proteins. miR-16 inhibition induced specific proteins associated with myogenic differentiation such as ACTA2, EEF1A2, and OPA1. We extend prior work in hypertrophic muscle tissue and show that miR-16 is lower in mechanically overloaded muscle in vivo. Our data collectively point to how miR-16 is implicated in aspects of myogenic cell differentiation. A deeper understanding of the role of miR-16 in myogenic cells has consequences for muscle developmental growth, exercise-induced hypertrophy, and regenerative repair after injury, all of which involve myogenic progenitors.

Automated summary

MicroRNA-16 (miR-16) plays a crucial role in stem cell biology and fate. It is lowly expressed in muscle during developmental hypertrophy and regeneration, and its inhibition enhances myoblast differentiation and myotube formation. Global transcriptomic and proteomic analyses revealed that miR-16 knockdown affects ribosomal protein gene expression, p53 pathway-related gene abundance, and regulation of tricarboxylic acid (TCA) cycle proteins and RNA metabolism-related proteins. miR-16 inhibition also induces specific proteins associated with myogenic differentiation. Overall, miR-16 is involved in myogenic cell differentiation.