Transcriptomics of mouse hepatocyte maturation and human hepatic reprogramming

Here, we chemically induced liver progenitor cells (CLiPs) from mouse hepatocytes using a previously established protocol. To identify central protein kinases (PKs) that potentially promote the maturation of lpc-Hep cells, we profiled the transcriptomes and phosphoproteomes of freshly isolated primary mouse hepatocytes (MHs) and ALBUMIN+ hepatocytes (CLiP-Heps) cells. Based on the algorithm developed by us and experiment verification, we validated three PKs that promote mouse hepatocyte maturation. Then, we use RNA sequencing technology to identify potential regulators during hepatic reprogramming with samples from HDFs 2.25 days (FHH-2.25d) and 5 days (FHH-5d) after FOXA3, HNF1A, and HNF4A (FHH) transduction, as well as HDFs transduced with GFP for 2.25 days (GFP) as the control group. After combined analysis with phosphoproteomic data with the algorithm and experiment verification, we identified 2 PKs that may be involved in hepatic reprogramming regulation. One of the five potential regulator candidates is PIM1, which can promote the progress remarkably with overexpression. Therefore, we next perform another RNA-seq to drive the molecular mechanisms under undergoing the regulation by PIM1. Finally, we revealed that cell cycle and ferroptosis pathways are involved in the regulation via PIM1. Overall design: For human hepatic reprogramming, mRNA profiles of HDFs 2.25 days (FHH-2.25d) and 5 days (FHH-5d) after FHH transduction and GFP cells, as well as HDFs undergoing hepatic conversion with PIM1 (PIM1-FHH) or GFP (GFP-FHH) overexpression. For mouse hepatocyte maturation, mRNA profiles of freshly isolated mouse primary hepatocytes (MH) and CLiP-Hep cells.