HNF4a nuclear localization correlates with the clinical progression of terminal hepatic failure in humans

HNF4a is a transcription factor that plays a critical role in terminal hepatocyte failure. HNF4a-based reprogramming therapy can correct terminal liver failure in rats and humans. As liver disease progresses, HNF4a expression decreases in the nuclei of hepatocytes, leading to impaired regulation and hepatic function. Post-translational modifications (PTMs) are a fundamental regulatory mechanism of protein function and localization. In this study, we analyzed HNF4a localization and pathways involved in HNF4a PTMs in human hepatocytes at different stages of decompensated liver function upon Child-Pugh classification. RNA-seq analysis revealed that HNF4a and the PTMs-pathway related to AKT are down-regulation in cirrhotic hepatocytes with terminal failure. These findings were confirmed by protein expression, where the HNF4a nuclear levels are significantly reduced in Child-Pugh B and C hepatocytes, whereas cytoplasmic expression of HNF4a was increased. Moreover, cMET and phospho-AKT were significantly reduced in Child-Pugh B and C hepatocytes. The association and statistical contribution of cMET and phospho-AKT to the nuclear localization of HNF4a were confirmed by Spearman's rank correlation test and pathway analysis. Principal component analysis was used to characterize the protein profiles related to the degree of liver dysfunction. Additionally, HNF4a acetylation was significantly reduced in failing human hepatocytes when compared to normal controls, demonstrating a significant correlation to the degree of hepatic function. Conclusion: These results suggest that the alterations in the cMET-AKT pathway directly correlate to HNF4a localization and the level of hepatic dysfunction. In conclusion, this study has therapeutic implications and suggests that manipulation of these pathways may restore hepatocyte function in terminal liver failure. Overall design: RNA-seq analysis of hepatocytes from normal patients, NASH patients, and alcohol-mediated Laennec's cirrhotic patients.