Transcriptome Analysis of Potential miRNA Involved in Adipogenic Differentiation of C2C12 Myoblasts

Abstract

>Emerging evidence indicates that microRNA (miRNA) plays an important role in adipogenesis and disease pathogenesis. To investigate the miRNA involved in regulating different periods of adipogenesis, we performed a comprehensive study on microRNAome during the stimulation of adipogenesis by an adipogenic differentiation cocktail in C2C12 myoblasts at 0, 2, 4, and 7 days using the Solexa sequencing technology. In this study, we identified 52 differentially expressed (DE) miRNA. Functional annotation indicated that the target genes of DE miRNA were mostly enriched in adipogenic transdifferentiation and fat metabolism-related pathways, including Wnt, mitogen-activated protein kinase (MAPK), and insulin signaling. The insulin signaling pathway was further analyzed for its close connections to Wnt and MAPK signaling pathways as well as for the containing of thymoma viral proto-oncogene-3 (Akt3) and glycogen synthase kinase-3 beta (Gsk3b), which were both target genes predicted by most DE miRNA. CLIP-seq (crosslinking-immunprecipitation and high-throughput sequencing) data showed that Akt3 was targeted by seven DE miRNA, including five upregulated (miR-203-3p, miR-181c-5p, miR-322-5p, miR-351-5p, and miR-181a-5p) and two downregulated (miR-15b-5p and miR-17-5p) ones. Likewise, Gsk3b was targeted by six DE miRNA, including four upregulated (miR-199a-3p, miR-126-5p, miR-26a-5p, and miR-101a-3p) and two downregulated (miR-150-5p and miR-140-3p) ones. Moreover, Akt3 could regulate the key transcription factor (TF) Foxo1, targeted by two downregulated miRNA (miR-96-5p and miR-183-5p). The expression levels of Akt3 and Gsk3b were downregulated, and TF Foxo1, which worked on the transcription axis of Pgc1a-Pparα-Rxrg-Pparγ to regulate adipogenesis, was upregulated. In conclusion, DE miRNA stimulated the adipogenesis of C2C12 myoblasts through the targeted insulin signaling pathway involving the genes Akt3, Gsk3b, and TF Foxo1.

Chromatin accessibility is associated with the changed expression of miRNAs that target members of the Hippo pathway during myoblast differentiation

“…Accumulating studies have shown that miRNAs are involved in many biological processes 8,40,41 , including skeletal myogenesis 10,16 . In this study, we performed a nonbiased analysis to assess miRNA and mRNA expression as well as ATAC-seq and ChIP-seq to investigate chromatin accessibility and TF occupancies, respectively.…”

“…Accumulating evidence has shown that muscle development is regulated by microRNAs (miR-NAs) in addition to the regulators described above. miRNAs have been reported to be involved in many biological processes, including skeletal myogenesis 6,7 and adipogenesis 8 . Muscle-specific miRNAs, including miR-1 9 , miR-133 10 , miR-206 11 , miR-208 12 , and miR-499 13 , have been considered key regulators with different targets during muscle development.…”

Genomic loci mispositioning inTmem120aknockout mice yields latent lipodystrophy

“…Of particular note, knockdown of miR-27b, miR-92a, miR-155, and miR-150 have all been shown to enhance browning/ beiging 60–63 , one of the Ad-Tmem120a -/- pathologies. Also, miR-322, miR-15b, miR-342-5p, and miR-16 are all pro-adipogenic 64–66 ; so their reduction is consistent with the pathology. Finally, miR-151-3p, miR-466f, miR-139-5p, miR-219, miR-29b, miR-25, and miR-370 are all linked to diabetes 67–73 , consistent with the inability to clear glucose, insulin resistance, and elevated insulin in Ad-Tmem120a -/- mice.…”

Potential Association of Circulating MicroRNA-181c and MicroRNA-484 Levels with Cardiorespiratory Fitness after Myocardial Infarction: A Pilot Study

“…35) miR-181c targeted the Akt-3 gene in cultured C2C12 myoblasts. 36) Akt-3 is a pro-survival signalling molecule in myoblasts. 37) Therefore, an intracellular increase of miR-181c in a myoblast may induce skeletal muscle dysfunction.…”