Fig 2: CKI-based analysis of hepatic reprogramming.a Experimental design of the trans-omics-based analysis of HDFs undergoing hepatic reprogramming by overexpression of FHH. b Heatmap of 15 potentially central PKs predicted by CKI using a threshold of =5 of 9 pairwise comparisons; the minus-log transformed p values were calculated for the indicated comparisons. c Expression levels of hepatic functional genes in HDFs overexpressing FHH and individual candidate central PKs for 5 days as quantified by qRT-PCR (n = 3). Overexpression of PIM1 (ALB p < 0.0001, AAT p < 0.0001, ARG p = 0.0020), PIM2 (ALB p = 0.0043, AAT p = 0.0071, ARG p = 0.0133), PIM3 (AAT p = 0.0022, ARG p < 0.0001), TRIB2 (ALB p = 0.0017), TSSK2 (ALB p = 0.0163, AAT p = 0.0027), TSSK3 (ALB p = 0.0077, AAT p = 0.0048), PSKH1 (AAT p = 0.0199), or CAMKV (AAT p = 0.0081) showed significantly changed transcript levels of hepatic functional genes. d Representative image of ALBUMIN immunofluorescence staining in HDFs overexpressing the indicated genes after infection with FHH for 12 days. Scale bars = 100 µm. n = 2 biological replicates. e Expression levels of hepatic functional genes in HDFs overexpressing FHH and shRNAs of candidate central PKs for 5 days as quantified by qRT-PCR (n = 3). Knockdown of PLK1 (ALB p = 0.0011, AAT p = 0.0104, ARG p = 0.0227), PLK2 (ALB p = 0.0038, AAT p = 0.0084, ARG p = 0.0010), PLK4 (ALB p = 0.0001, AAT p = 0.0070), PIM1 (ALB p = 0.0006, AAT p = 0.0113, ARG p = 0.0002), PIM2 (ALB p = 0.0398, AAT p = 0.0010, ARG p = 0.0222), TRIB2 (ALB p = 0.0392), ROCK2 (ALB p = 0.0125, ARG p = 0.0010), or TSSK3 (AAT p = 0.0459, ARG p = 0.0101) showed significantly changed transcript levels of hepatic functional genes. Data are shown as the mean + SD. *p < 0.05, **p < 0.01, ***p < 0.001 (unpaired two-sided Student’s t-test). Source data are provided as a Source Data file.

Fig 3: Proposed signaling pathway that inhibits endothelial cell angiogenesis under high glucose and low serum conditions. miR, microRNA; PIM1, pim-1 proto-oncogene, serine/threonine kinase; p, phosphorylated; VEGF, vascular endothelial growth factor.

Fig 4: PIM1 suppresses hepatic reprogramming-induced ferroptosis.a Representative image of FHH-induced cell death of HDFs on day 5. Scale bars = 100 µm. n = 4 biological replicates. b FHH-induced cell death was analyzed by Annexin V/7-AAD staining on day 5. Proportions of 7-AAD/Annexin V positive cells were quantified (p < 0.0001, n = 5). c Expression levels of ferroptosis-related genes were increased on day 5 of hepatic transdifferentiation (SLC7A11 p = 0.0009, PTGS2 p = 0.0007, ACSL4 p = 0.0013, HMOX1 p < 0.0001, n = 3). d Knockdown of PIM1 increased the transcript levels of ferroptosis genes on day 5 of hepatic transdifferentiation (SLC7A11 p = 0.0015, PTGS2 p = 0.0002, ACSL4 p = 0.0121, HMOX1 p = 0.0002, n = 3). e Overexpression of PIM1 decreased the transcript levels of ferroptosis genes on day 5 of hepatic transdifferentiation (SLC7A11 p = 0.0062, PTGS2 p = 0.0011, ACSL4 p = 0.0025, HMOX1 p = 0.0080, n = 3). The samples were the same with those used in Fig. 4e, f. f Propidium iodide (PI) staining assay showing that treatment with the ferroptosis inhibitors ferrostatin-1 and liproxstatin-1 inhibited FHH-induced cell death in HDFs. Scale bars = 100 µm. n = 2 biological replicates. g Treatment of NAC, Fer-1, or Lip-1 increased the transcript levels of liver-specific genes on day 5 of hepatic transdifferentiation (NAC treatment: ALB p = 0.0041, CYP3A4 p = 0.0004; Fer-1 treatment: ALB p = 0.0008, CYP3A4 p = 0.0001; Lip-1 treatment: ALB p = 0.0198, CYP3A4 p = 0.0404; n = 3). h Cellular NADP/NADPH levels on day 3 of hepatic transdifferentiation with GFP or PIM1 overexpression (NADPH p = 0.0015, NADPH/NADP+ p = 0.0041, n = 3). i Cellular GSH levels on day 5 of hepatic transdifferentiation with GFP or PIM1 overexpression (p = 0.0002, n = 3). Data are shown as the mean + SD. *p < 0.05, **p < 0.01, ***p < 0.001 (unpaired two-sided Student’s t-test). Source data are provided as a Source Data file.

Fig 5: Network analysis of the major molecular landscape in hepatic reprogramming.a An integrative TPCW based on the findings of this study and existing knowledge illustrates the regulatory relationships among FHH, 24 FHH-regulated TFs, two screened central PKs, and 60 curated genes related to hepatic lineage, ferroptosis, and cell cycle progression. b PIM1-centered subnetwork during hepatic lineage reprogramming. c Working model showing potential roles of PIM1 in hepatic lineage reprogramming.