Liver‐enriched transcription factor expression relates to chronic hepatic failure in humans

Guzman‐Lepe, Jorge; Cervantes-Álvarez, Eduardo; l’Hortet, Alexandra Collin de; Wang, Yan; Mars, Wendy M.; Oda, Yoshinao; Bekki, Yuki; Shimokawa, Masahiro; Wang, Huanlin; Yoshizumi, Tomoharu; Maehara, Yoshihiko; Bell, Aaron; Fox, Ira J.; Takeishi, Kazuki; Soto-Gutiérrez, Alejandro

doi:10.1002/hep4.1172

Cited by 31 publications

(48 citation statements)

References 52 publications

(112 reference statements)

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“…This result was not surprising, as the ability to discern a difference in HNF4α expression in livers from patients with degenerative disease and controls based on degree of functional decompensation required the study of a large cohort of patients. ( 5 ) However, in this study, a statistically significant difference was observed based on HNF4α location. HNF4α was detected at high levels in the cytoplasm ( P = 0.023; Fig.…”

Section: Resultscontrasting

confidence: 65%

“…( 12 ) To assess whether this observation applies to humans, we studied liver‐enriched transcription factor expression in the livers of a large cohort of patients with decompensated liver function. ( 5 ) We found that HNF4α mRNA levels were down‐regulated and correlated with the extent of liver dysfunction based on Child‐Pugh classification. Interestingly, in those studies, nuclear localization of HNF4α was not uniform.…”

Section: Discussionmentioning

confidence: 81%

“…There are numerous causes of chronic liver disease, including chronic infection by hepatitis viruses, alcohol‐mediated cirrhosis, and nonalcoholic steatohepatitis (NASH), ( 4 ) and each can produce hepatocellular failure. ( 5,6 ) The mechanisms responsible for deterioration of hepatocyte function and ultimately hepatic failure in man are poorly understood. Chronic hepatic damage produces oxidative stress ( 7 ) and endoplasmic stress, ( 8 ) which can induce cell death ( 8‐10 ) and reduce the proliferative capacity of the hepatocytes.…”

mentioning

confidence: 99%

“…( 12 ) We have also shown in a large cohort of patients with advanced liver disease that the level of HNF4α messenger RNA (mRNA) expression in the diseased liver correlated with extent of hepatic dysfunction (Childs‐Pugh classification) and that HNF4α expression was not localized in the nucleus, as was the case in our rat studies. ( 5 )…”

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confidence: 99%

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Cellular Location of HNF4α is Linked With Terminal Liver Failure in Humans

et al. 2020

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Hepatocyte nuclear factor 4 alpha (HNF4α) is a transcription factor that plays a critical role in hepatocyte function, and HNF4α-based reprogramming corrects terminal liver failure in rats with chronic liver disease. In the livers of patients with advanced cirrhosis, HNF4α RNA expression levels decrease as hepatic function deteriorates, and protein expression is found in the cytoplasm. These findings could explain impaired hepatic function in patients with degenerative liver disease. In this study, we analyzed HNF4α localization and the pathways involved in post-translational modification of HNF4α in human hepatocytes from patients with decompensated liver function. RNA-sequencing analysis revealed that AKT-related pathways, specifically phospho-AKT, is down-regulated in cirrhotic hepatocytes from patients with terminal failure, in whom nuclear levels of HNF4α were significantly reduced, and cytoplasmic expression of HNF4α was increased. cMET was also significantly reduced in failing hepatocytes. Moreover, metabolic profiling showed a glycolytic phenotype in failing human hepatocytes. The contribution of cMET and phospho-AKT to nuclear localization of HNF4α was confirmed using Spearman's rank correlation test and pathway analysis, and further correlated with hepatic dysfunction by principal component analysis. HNF4α acetylation, a posttranslational modification important for nuclear retention, was also significantly reduced in failing human hepatocytes when compared with normal controls. Conclusion: These results suggest that the alterations in the cMET-AKT pathway directly correlate with HNF4α localization and level of hepatocyte dysfunction. This study suggests that manipulation of HNF4α and pathways involved in HNF4α posttranslational modification may restore hepatocyte function in patients with terminal liver failure. (Hepatology Communications 2020;4:859-875).

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Section: Resultscontrasting

confidence: 65%

Section: Discussionmentioning

confidence: 81%

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confidence: 99%

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Cellular Location of HNF4α is Linked With Terminal Liver Failure in Humans

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“…Top 50 transcription factors revealed major changes in liver-specific transcription factors (HNF4α, C/ EBP, EGR1, and FOXA1) ( Fig. 1d; Supplemental Table 2) (Guerquin et al 2013;Guzman-lepe et al 2018). For the top five TFs, we observed large down-regulated regulons for E2F4 and ETS1 (mainly involved in cell cycle) (Hsu and Sage 2016;Fry and Inoue 2018) with suppressed activity over time, and large up-regulated regulons for HNF4α, C/EBP, and STAT1 (mainly involved in differentiation) with increased activation over time ( Fig.…”

Section: Transcriptome Analysis Of 3d Spheroid Hepg2 Differentiationmentioning

confidence: 99%

High-throughput confocal imaging of differentiated 3D liver-like spheroid cellular stress response reporters for identification of drug-induced liver injury liability

et al. 2019

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Adaptive stress response pathways play a key role in the switch between adaptation and adversity, and are important in druginduced liver injury. Previously, we have established an HepG2 fluorescent protein reporter platform to monitor adaptive stress response activation following drug treatment. HepG2 cells are often used in high-throughput primary toxicity screening, but metabolizing capacity in these cells is low and repeated dose toxicity testing inherently difficult. Here, we applied our bacterial artificial chromosome-based GFP reporter cell lines representing Nrf2 activation (Srxn1-GFP and NQO1-GFP), unfolded protein response (BiP-GFP and Chop-GFP), and DNA damage response (p21-GFP and Btg2-GFP) as long-term differentiated 3D liver-like spheroid cultures. All HepG2 GFP reporter lines differentiated into 3D spheroids similar to wild-type HepG2 cells. We systematically optimized the automated imaging and quantification of GFP reporter activity in individual spheroids using high-throughput confocal microscopy with a reference set of DILI compounds that activate these three stress response pathways at the transcriptional level in primary human hepatocytes. A panel of 33 compounds with established DILI liability was further tested in these six 3D GFP reporters in single 48 h treatment or 6 day daily repeated treatment. Strongest stress response activation was observed after 6-day repeated treatment, with the BiP and Srxn1-GFP reporters being most responsive and identified particular severe-DILI-onset compounds. Compounds that showed no GFP reporter activation in two-dimensional (2D) monolayer demonstrated GFP reporter stress response activation in 3D spheroids. Our data indicate that the application of BAC-GFP HepG2 cellular stress reporters in differentiated 3D spheroids is a promising strategy for mechanism-based identification of compounds with liability for DILI.

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Hepatocyte Nuclear Factor 4 alpha 2 Messenger RNA Reprograms Liver‐Enriched Transcription Factors and Functional Proteins in End‐Stage Cirrhotic Human Hepatocytes

et al. 2021

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The only definitive therapy for end-stage liver disease is whole-organ transplantation. The success of this intervention is severely limited by the complexity of the surgery, the cost of patient care, the need for long-term immunosuppression, and the shortage of donor organs. In rodents and humans, end-stage degeneration of hepatocyte function is associated with disruption of the liver-specific transcriptional network and a nearly complete loss of promoter P1-driven hepatocyte nuclear factor 4-alpha (P1-HNF4α) activity. Re-expression of HNF4α2, the predominant P1-HNF4α, reinstates the transcriptional network, normalizes the genes important for hepatocyte function, and reverses liver failure in rodents. In this study, we tested the effectiveness of supplementary expression of human HNF4α2 messenger RNA (mRNA) in primary human hepatocytes isolated from explanted livers of patients who underwent transplant for end-stage irreversibly decompensated liver failure (Child-Pugh B, C) resulting from alcohol-mediated cirrhosis and nonalcoholic steatohepatitis. Re-expression of HNF4α2 in decompensated cirrhotic human hepatocytes corrects the disrupted transcriptional network and normalizes the expression of genes important for hepatocyte function, improving liver-specific protein expression. End-stage liver disease in humans is associated with both loss of P1-HNF4α expression and failure of its localization to the nucleus. We found that while HNF4α2 re-expression increased the amount of P1-HNF4α protein in hepatocytes, it did not alter the ability of hepatocytes to localize P1-HNF4α to their nuclei. Conclusion: Re-expression ofHNF4α2 mRNA in livers of patients with end-stage disease may be an effective therapy for terminal liver failure that would circumvent the need for organ transplantation. The efficacy of this strategy may be enhanced by discovering the cause for loss of nuclear P1-HNF4α localization in end-stage cirrhosis, a process not found in rodent studies. (Hepatology Communications 2021;0:1-16).O ver 1 million deaths are estimated per year worldwide due to complications of endstage liver disease. (1) The most common causes include chronic infection by hepatitis viruses, alcohol-mediated cirrhosis, and nonalcoholic steatohepatitis (NASH); the final step in decompensated disease is the development of portal hypertension and hepatocellular failure. (2) Pathogenesis involves

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Liver‐enriched transcription factor expression relates to chronic hepatic failure in humans

Cited by 31 publications

References 52 publications

Cellular Location of HNF4α is Linked With Terminal Liver Failure in Humans

Cellular Location of HNF4α is Linked With Terminal Liver Failure in Humans

High-throughput confocal imaging of differentiated 3D liver-like spheroid cellular stress response reporters for identification of drug-induced liver injury liability

Hepatocyte Nuclear Factor 4 alpha 2 Messenger RNA Reprograms Liver‐Enriched Transcription Factors and Functional Proteins in End‐Stage Cirrhotic Human Hepatocytes

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