Pharmacologic augmentation of γ-
globin expression sufficient to reduce
anemia and clinical severity in patients with diverse
hemoglobinopathies has been challenging. In studies here, representative molecules from four chemical classes, representing several distinct primary mechanisms of action, were investigated for effects on γ-
globin transcriptional repressors, including components of the
NuRD complex (LSD1 and HDACs 2-3), and the downstream repressor BCL11A, in erythroid progenitors from
hemoglobinopathy patients. Two
HDAC inhibitors (
MS-275 and SB939), a
short-chain fatty acid derivative (
sodium dimethylbutyrate [
SDMB]), and an agent identified in high-throughput screening,
Benserazide, were studied. These
therapeutics induced γ-
globin mRNA in progenitors above same subject controls up to 20-fold, and increased F-reticulocytes up to 20%. Cellular
protein levels of BCL11A, LSD-1, and KLF1 were suppressed by the compounds.
Chromatin immunoprecipitation assays demonstrated a 3.6-fold reduction in LSD1 and HDAC3 occupancy in the γ-
globin gene promoter with
Benserazide exposure, 3-fold reduction in LSD-1 and HDAC2 occupancy in the γ-
globin gene promoter with
SDMB exposure, while markers of gene activation (
histone H3K9 acetylation and H3K4 demethylation), were enriched 5.7-fold. These findings identify clinical-stage oral
therapeutics which inhibit or displace major
co-repressors of γ-
globin gene transcription and may suggest a rationale for combination
therapy to produce enhanced efficacy.