CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, P. R. China.
CAS Key Laboratory of Computational Biology, CAS-MPG Partner Institute for Computational Biology, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, P. R. China.
Stem Cell Reports. 2018 Jul 10;11(1):22-31. doi: 10.1016/j.stemcr.2018.05.001. Epub 2018 May 31.
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 comprise a heterogeneous population. Thus, a better understanding of HLC differentiation will improve the likelihood of future application. Here, by taking advantage of CRISPR-Cas9-based genome-wide screening technology and a high-throughput hPSC screening platform with a reporter readout, we identified several potential genetic regulators of HLC differentiation. By using a chemical screening approach within 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. Mechanistically, 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 optimizing hPSC differentiation.
人多能干细胞（hPSC）来源的肝细胞样细胞（HLC）为疾病建模和移植提供了有前途的细胞资源。然而，分化的 HLC 表现出不成熟的表型，并包含异质群体。因此，更好地了解 HLC 分化将提高未来应用的可能性。在这里，我们利用基于 CRISPR-Cas9 的全基因组筛选技术和具有报告基因读出的高通量 hPSC 筛选平台，鉴定了几个潜在的 HLC 分化的遗传调控因子。通过在我们的平台内进行化学筛选，我们还鉴定出了可以进一步促进 HLC 分化并保留体外培养原代肝细胞特征的化合物。值得注意的是，这两种筛选都将组蛋白去乙酰化酶 3（HDAC3）鉴定为肝分化中的关键调控因子。在机制上，HDAC3 与肝转录因子（如 HNF4）形成复合物，并在肝分化过程中共同调控转录程序。这项研究强调了一种广泛适用于研究和优化 hPSC 分化的方法。
