HDAC8 regulates neural differentiation through embryoid body formation in P19 cells

Highlights: • Selective HDAC8 inhibitor causes reduced embryoid body (EB) formation in P19 cells. • HDAC8-inhibited EBs show decreased neural differentiation activity. • HDAC8 inhibition leads to G2/M phase accumulation. • HDAC8 inhibition causes downregulation of cyclin A2 and cyclin B1 gene expression. • CRISPR-Cas9-mediated HDAC8 knockout cells show smaller EBs. Histone acetylation and deacetylation correlate with diverse biological phenomena through gene transcription. Histone deacetylases (HDACs) regulate deacetylation of histones and other proteins. However, as a member of the HDAC family, HDAC8 function during neurodevelopment is currently unknown. Therefore, we investigated HDAC8 function during neurodevelopment by examining embryoid body (EB) formation in P19 cells. HDAC8-selective inhibitor (NCC-149) (HDAC8i)-treated cells showed smaller EBs than non-treated cells, as well as reduced expression levels of the neuronal marker, NeuN. Additionally, HDAC8i treatment led to inhibition of cellular proliferation by G2/M phase accumulation and downregulated cyclin A2 and cyclin B1 gene expression. Furthermore, two independent HDAC8 knockout cell lines were established by CRISPR-Cas9, which resulted in smaller EBs, similar to HDAC8i-treated cells. These results suggest that HDAC8 regulates neural differentiation by exerting control of EB formation.