TGF-β Induction of miR-143/145 Is Associated to Exercise Response by Influencing Differentiation and Insulin Signaling Molecules in Human Skeletal Muscle

Physical training improves insulin sensitivity and can prevent type 2 diabetes (T2D). However, approximately 20% of individuals lack a beneficial outcome in glycemic control. TGF-beta, identified as a possible upstream regulator involved in this low response, is also a potent regulator of microRNAs (miRNAs). The aim of this study was to elucidate the potential impact of TGF-beta-driven miRNAs on individual exercise response. Non-targeted long and sncRNA sequencing analyses of TGF-beta 1-treated human skeletal muscle cells corroborated the effects of TGF-beta 1 on muscle cell differentiation, the induction of extracellular matrix components, and identified several TGF-beta 1-regulated miRNAs. qPCR validated a potent upregulation of miR-143-3p/145-5p and miR-181a2-5p by TGF-beta 1 in both human myoblasts and differentiated myotubes. Healthy subjects who were overweight or obese participated in a supervised 8-week endurance training intervention (n = 40) and were categorized as responder or low responder in glycemic control based on fold change ISIMats (& GE;+1.1 or <+1.1, respectively). In skeletal muscle biopsies of low responders, TGF-beta signaling and miR-143/145 cluster levels were induced by training at much higher rates than among responders. Target-mining revealed HDACs, MYHs, and insulin signaling components INSR and IRS1 as potential miR-143/145 cluster targets. All these targets were down-regulated in TGF-beta 1-treated myotubes. Transfection of miR-143-3p/145-5p mimics in differentiated myotubes validated MYH1, MYH4, and IRS1 as miR-143/145 cluster targets. Elevated TGF-beta signaling and miR-143/145 cluster induction in skeletal muscle of low responders might obstruct improvements in insulin sensitivity by training in two ways: by a negative impact of miR-143-3p on muscle cell fusion and myofiber functionality and by directly impairing insulin signaling via a reduction in INSR by TGF-beta and finetuned IRS1 suppression by miR-143-3p.

Automated summary

This study investigated the impact of TGF-beta on individual exercise response, revealing its role in regulating miRNA and observing significant induction of TGF-beta signaling and the miR-143/145 cluster in non-responders. The findings suggested that elevated TGF-beta signaling and miR-143/145 cluster induction in skeletal muscle of non-responders may hinder improvements in insulin sensitivity by affecting muscle cell fusion, myofiber functionality, and insulin signaling components.