GEOTranscriptomicsMus musculusExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE213423GEOGSEfalseRole of TAp63 in myogenesis [RNA-Seq]Myogenesis is an ordered process driven by myogenic regulatory factors (MyoD, MyoG, MRF4) culminating in the fusion of myoblasts in myotubes. p53 family members are involved in myoblasts differentiation, although the role of the specific isoforms has not been investigated. Previous experiments indicated that TAp63g isoform is the only member of the p53 family up-regulated during in vitro myogenesis, suggesting an important function for this specific isoform during this process. As experimental models we used stable modified murine myoblasts cell lines C2C7 and C2C12 (shp63 C2C7 and doxycyclin-inducible TAp63g C2C12). Microarray analysis of shp63 C2C7 cells compared to control revealed changes in the expression of genes which codify for proteins involved in cellular proliferation and metabolic processes. We confirmed that shp63 clones had an increase in their proliferative potential, evaluated by clonogenicity assay and EdU incorporation assay, and a delay in differentiation, evaluated by WB analysis of specific differentiation markers. The same assays showed that TAp63g overexpression is able to decrease cell proliferation. Moreover, we observed an increase in mitochondrial ROS production evaluated by citofluorimetric techniques and a decrease in oxygen consumption, revealed by Seahorse analyser. RNA sequencing and ChIP sequencing for histone modification H3K27ac, performed during C2C7 differentiation, followed by microarray analysis carried out in shp63 cells revealed also several genes (coding and non coding) modulated during in vitro myoblasts differentiation. Among them, we found Igf2. Igf2 is involved in muscle development and differentiation, both in vivo and in vitro. We found that p63 binds its enhancer regions, indicating its involvement in regulating Ifg2 expression during differentiation. Interestingly, we found that the long non coding RNA, Airn, is down-regulated in absence of p63 during differentiation and its silencing led to a decrease in either mRNA and protein level of myogenic differentiation markers (MyoD and MyoG). Moreover, by luciferase activity assay, we found that p63 regulates Airn expression directly, binding specific enhancers. In summary, our data reveal a novel role for TAp63 in controlling myoblasts proliferation, energy metabolism and also differentiation through the expression of specific target protein coding genes and long non coding genes. Preliminary observations, using publicity available datasets, confirmed that TAp63 isoform is among the p53-family members the isoform more expressed in adult human normal skeletal muscle. Interestingly, a genome-wide gene expression profiling of skeletal muscle from early Duchenne muscular dystrophy (DMD) patients showed a significant reduction of the TAp63 isoform expression in the affected tissues. The latter observation indicate new possible scenario for studying TAp63 contribution to muscle degenerative pathologies.2026/06/03GSE213423GSM6586000GSM6586001GSM6586002GSM6586003GSM6586004GSM658600519057213423Mus musculus[42210377]