RNA-seq transcriptomic analysis of 4-octyl itaconate repressing myogenic differentiation

Area of Science:

• Molecular Biology
• Cell Biology
• Biochemistry

Background:

• 4-octyl itaconate (OI) is a cell-permeable itaconate derivative with known anti-fibrotic effects.
• OI has been shown to influence osteoclast differentiation.
• The impact of OI on myoblast differentiation requires further elucidation.

Purpose of the Study:

• To investigate the molecular mechanisms by which 4-octyl itaconate (OI) affects myoblast differentiation.
• To utilize RNA-sequencing (RNA-seq) analysis to explore these underlying mechanisms.

Main Methods:

• C2C12 myoblasts were treated with varying concentrations of OI (2.5–100 μmol/L).
• Myoblast proliferation, differentiation markers (myosin heavy chain, myogenin), and muscle regulatory factors were assessed.
• RNA-sequencing, gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene set enrichment analysis (GSEA) were employed.

Main Results:

• OI did not affect myoblast proliferation but inhibited differentiation in a dose-dependent manner.
• OI suppressed the expression of myosin heavy chain (MHC) and myogenin.
• OI inhibited myogenic differentiation by affecting MyoD-regulated activity via AKT1 phosphorylation inhibition, impacting PI3K-Akt, calcium, and PPAR signaling pathways.

Conclusions:

• This study elucidates the molecular mechanisms of OI's inhibition of myogenic differentiation.
• OI targets multiple molecules and pathways, including PI3K-Akt signaling, to exert its effects.
• Findings provide novel insights into the multifaceted actions of OI in myogenesis.