Genetic and chemical screenings identify HDAC3 as a key regulator in hepatic differentiation of human pluripotent stem cells

Hepatocyte-like cells (HLCs) derived from human pluripotent stem cells (hPSCs) offer a promising cell resource for disease modeling and transplantation. However, differentiated HLCs exhibit an immature phenotype and remain a heterogeneous population. To improve the quality of HLCs, we took advantage of CRISPR-Cas9-based genome-wide screening technology and developed a high-throughput hPSC screening platform with a reporter readout. We have identified several potential genetic regulators of the differentiation process. When we applied a chemical screening approach to our platform, we also identified compounds that can further promote HLC differentiation and preserve the characteristics of in vitro cultured primary hepatocytes. Remarkably, both screenings identified histone deacetylase 3 (HDAC3) as a key regulator in hepatic differentiation. Further studies indicated that HDAC3 formed a complex with liver transcriptional factors, e.g., HNF4, and co-regulated the transcriptional program during hepatic differentiation. This study highlights a broadly useful approach for studying and the optimization of hPSC differentiation.