Histone Deacetylase 9 Activates γ-Globin Gene Expression in Primary Erythroid Cells

Strategies to induce fetal hemoglobin (HbF) synthesis for the treatment of beta-hemoglobinopathies probably involve protein modifications by histone deacetylases (HDACs) that mediate gamma-globin gene regulation. However, the role of individual HDACs in globin gene expression is not very well understood; thus, the focus of our study was to identify HDACs involved in gamma-globin activation. K562 erythroleukemia cells treated with the HbF inducers hemin, trichostatin A, and sodium butyrate had significantly reduced mRNA levels of HDAC9 and its splice variant histone deacetylase-related protein. Subsequently, HDAC9 gene knockdown produced dose-dependent gamma-globin gene silencing over an 80-320 nM range. Enforced expression with the pTarget-HDAC9 vector produced a dose-dependent 2.5-fold increase in gamma-globin mRNA (p < 0.05). Furthermore, ChIP assays showed HDAC9 binding in vivo in the upstream G gamma-globin gene promoter region. To determine the physiological relevance of these findings, human primary erythroid progenitors were treated with HDAC9 siRNA; we observed 40 and 60% gamma-globin gene silencing in day 11 (early) and day 28 (late) progenitors. Moreover, enforced HDAC9 expression increased gamma-globin mRNA levels by 2.5-fold with a simultaneous 7-fold increase in HbF. Collectively, these data support a positive role for HDAC9 in gamma-globin gene regulation.