In vivo CRISPR screen reveals interplay of epigenetic regulators underlies kinetics of hepatobiliary reprogramming [RNA-Seq]

Under chronic liver injury, a small fraction of hepatocytes undergoes reprogramming to biliary epithelial cells (BECs) through a multistep process of chromatin and transcriptional remodeling in which the hepatocyte phenotype is repressed and the biliary phenotype is activated. However, the epigenetic basis underlying this phenomenon is largely unknown. Here, we profiled cells after knockout of Nsd1 or Kdm2a at two reprogramming stages to assess the transcriptional differences imposed by epigenetic dysregulation. Mice were challenged with DDC for 8 weeks, then reprogrammed cells and biliary cells were harvested. After DDC-induced cholestatic injury, GFP+CD24- cells (DDC-injured hepatocytes/early reprogrammed cells) and GFP+CD24+ cells were isolated, including the intermediate and late stages of reprogrammed hepatocytes, from Kdm2a-KO, Nsd1-KO, or empty vector (EV) control mice.