The inhibitory effects of Remodelin on murine myoblasts differentiation

Abstract

Myoblasts differentiation is a highly regulated and complex process leading to the formation of fused and aligned mature myotubes. Growing interest in the role of epigenetics in muscle differentiation has highlighted epi-modulators as crucial regulators of this process. Our in vitro study aimed to explore the potential effects of the inhibition of the acetyltransferase Nat10 on myoblasts differentiation, by using Remodelin, a Nat10 selective inhibitor. We cultivated and differentiated murine C2C12 myoblasts on ultra-compliant gelatin substrates for up to 16 days and treated them with Remodelin. A combination of morphological analyses, confocal microscopy, transcriptomic profiling (RNA-seq), quantitative proteomics and metabolomics analyses was employed to assess the impact of Nat10 inhibition on myotube formation and maturation. To evaluate the reproducibility of Remodelin effects across myogenic systems and species, L6 rat myoblasts were included as a secondary comparative model. Remodelin treatment impaired myotube organization, alignment, and structural maturation in both C2C12 and L6 cells compared to untreated controls. In C2C12 cultures, Remodelin also abolished spontaneous myotube contractility. Intersection of transcriptomics and proteomics analyses confirmed that Remodelin effectively slowed myotube formation. Overall, these results indicate that Remodelin broadly affects the regulatory networks involved in skeletal muscle differentiation.