Targeted modulation of FGFR1 enhances myogenic differentiation for cultured meat production

Abstract

The development of cultured meat offers a sustainable and ethical alternative to conventional protein sources, yet its commercial scalability is hindered by the inefficient induction of terminal myogenesis. Porcine muscle stem cells (PMSCs) are a promising cell source, but their effective differentiation into mature muscle fibers remains a significant challenge. The fibroblast growth factor receptor 1 (FGFR1) plays a pivotal role in maintaining the proliferative state of these cells, making its inhibition a compelling strategy to promote differentiation. In this study, we employed a multi-faceted approach, combining in silico and in vitro methods, to identify and validate novel small-molecule FGFR1 modulators. A rigorous virtual screening of 872 compounds against the FGFR1 tyrosine kinase domain, with subsequent filtering against the off-target p38α MAPK, identified methylergometrine as a highly promising candidate alongside the known inhibitor dovitinib. Molecular dynamics simulations confirmed that both compounds form stable complexes with FGFR1, a finding corroborated by saturation transfer difference-nuclear magnetic resonance, which provided direct evidence of their distinct but complementary binding modes. Subsequent in vitro functional assays in C2C12 mouse myogenic cells demonstrated that both compounds enhance myogenic differentiation. Specifically, treatment with dovitinib and methylergometrine led to a robust, dose- and phase-dependent upregulation of key myogenic markers, including myosin heavy chain and myogenin. These findings confirm that dovitinib and methylergometrine effectively promote myotube formation. Similar trends were also observed in PMSCs. Our study introduces a novel strategy for stimulating terminal myogenesis through targeted FGFR1 modulation, with significant implications for improving the efficiency and yield of cell-based cultured meat production.