Reduced Dnmt3a increases Gdf5 expression with suppressed satellite cell differentiation and impaired skeletal muscle regeneration

Abstract

DNA methylation is an epigenetic mechanism regulating gene expression. In this study, we observed that DNA methyltransferase 3a (Dnmt3a) expression is decreased after muscle atrophy. We made skeletal muscle-specific Dnmt3a-knockout (Dnmt3a-KO) mice. The regeneration capacity after muscle injury was markedly decreased in Dnmt3a-KO mice. Diminished mRNA and protein expression of Dnmt3a were observed in skeletal muscles as well as in satellite cells, which are important for muscle regeneration, in Dnmt3a-KO mice. Dnmt3a-KO satellite cell showed smaller in size (length/area), suggesting suppressed myotube differentiation. Microarray analysis of satellite cells showed that expression of growth differentiation factor 5 (Gdf5) mRNA was markedly increased in Dnmt3a-KO mice. The DNA methylation level of the Gdf5 promoter was markedly decreased in Dnmt3a-KO satellite cells. In addition, DNA methylation inhibitor azacytidine treatment increased Gdf5 expression in wild-type satellite cells, suggesting Gdf5 expression is regulated by DNA methylation. Also, we observed increased inhibitor of differentiation (a target of Gdf5) mRNA expression in Dnmt3a-KO satellite cells. Thus, Dnmt3a appears to regulate satellite cell differentiation via DNA methylation. This mechanism may play a role in the decreased regeneration capacity during atrophy such as in aged sarcopenia.-Hatazawa, Y., Ono, Y., Hirose, Y., Kanai, S., Fujii, N. L., Machida, S., Nishino, I., Shimizu, T., Okano, M., Kamei, Y., Ogawa, Y. Reduced Dnmt3a increases Gdf5 expression with suppressed satellite cell differentiation and impaired skeletal muscle regeneration.

Effect of COL5A1, GDF5, and PPARA Genes on a Movement Screen and Neuromuscular Performance in Adolescent Team Sport Athletes

“…Especially as PPARs and their coactivators are associated with improvements in training programs for weight reduction (29), aerobic performance (49,52,60,61) and resistance training load capabilities (1). In this manner, the ketogenesis and other metabolic factors determined by PPARA indicate an individual response to strength and power training (1), and satellite cell proliferation determined by GDF5 can indicate a potential to regenerate from a long term physical load (18). Regarding the fact that the interaction of these genes in terms of performance, injury prevention and fatigue factors was not analysed, the authors suggest that this analysis should be performed in future studies.…”

“…Peroxisome proliferator-activated receptor alpha (PPARA) and growth differentiation factor (GDF5) genes have been associated with power performance (48), stress fractures (66) and muscle regeneration (18). Specifically, it has been suggested that PPARA predicts anaerobic trainability (1), and aerobic trainability (43,47,49), while GDF5 regulates the response of the proliferation satellite cell (18) (crucial after resistance training), meniscus injury incidents, and knee joint function recovery (14).…”

Analysis of Genomic DNA Methylation and Gene Transcription Modulation Induced by DNMT3A Deficiency in HEK293 Cells

“…Several previous studies have focused on DNMT3A gene knockout in human or mouse-resourced cells, including human embryotic stem cells, human leukemia cells K562, mouse hematopoietic stem cells, as well as mouse somatic cells [2,14,27,28]. Compared to mouse cells, human cells are less tolerant to DNMT3A de ciency and it can cause lethality and genomic instability in the cells.…”

Analysis of genomic DNA methylation and gene transcription modulation induced byDNMT3Adeficiency in HEK293 cells

“…Several previous studies have focused on DNMT3A gene knockout in human or mouse-resourced cells, including human embryotic stem cells, human leukemia cells K562, mouse hematopoietic stem cells, as well as mouse somatic cells (Banaszak et al, 2018;Hatazawa et al, 2018;Jeong et al, 2018;Liao et al, 2015). Compared to mouse cells, human cells are less tolerant to DNMT3A deficiency and it can cause lethality and genomic instability in the cells.…”