Convergence of Wnt Signaling on the HNF4α-Driven Transcription in Controlling Liver Zonation

Alice Conigliaro, Alice Conigliaro, Laura Santangelo, Emiliano Pasquini, Marta Colletti, Laura Amicone, Carla Cicchini, Tonino Alonzi, Marco Tripodi

Risultato della ricerca: Article

90 Citazioni (Scopus)

Abstract

Background & Aims: In each hepatocyte, the specific repertoire of gene expression is influenced by its exact location along the portocentrovenular axis of the hepatic lobule and provides a reason for the liver functions compartmentalization defined "metabolic zonation." So far, few molecular players controlling genetic programs of periportal (PP) and perivenular (PV) hepatocytes have been identified; the elucidation of zonation mechanisms remains a challenge for experimental hepatology. Recently, a key role in induction and maintenance of the hepatocyte heterogeneity has been ascribed to Wnt/β-catenin pathway. We sought to clarify how this wide-ranging stimulus integrates with hepatocyte specificity. Methods: Reverse transcriptase polymerase chain reaction (RT-PCR) allowed the transcriptional profiling of hepatocytes derived from in vitro differentiation of liver stem cells. The GSK3β inhibitor 6-bromoindirubin-3â²-oxime (BIO) was used for β-catenin stabilization. Co-immunoprecipitations were used to study biochemical protein interactions while ChIP assays allowed the in vivo inspection of PV and PP genes regulatory regions. Results: We found that spontaneous differentiation of liver stem cells gives rise to PP hepatocytes that, after Wnt pathway activation, switch into PV hepatocytes. Next, we showed that the Wnt downstream player LEF1 interacts with the liver-enriched transcriptional factor HNF4α. Finally, we unveiled that the BIO induced activation of PV genes correlates with LEF1 binding to both its own and HNF4α consensus, and the repression of PP genes correlates with HNF4α displacement from its own consensus. Conclusion: Our data show a direct and hitherto unknown convergence of the canonical Wnt signaling on the HNF4α-driven transcription providing evidences of a mechanism controlling liver zonated gene expression. © 2009 AGA Institute.
Lingua originaleEnglish
pagine (da-a)660-672
Numero di pagine13
RivistaGastroenterology
Volume137
Stato di pubblicazionePublished - 2009

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Hepatocytes
Liver
Catenins
Wnt Signaling Pathway
Stem Cells
Gene Expression
Nucleic Acid Regulatory Sequences
Gastroenterology
Reverse Transcriptase Polymerase Chain Reaction
Immunoprecipitation
Transcriptional Activation
Genes
Maintenance
Proteins

All Science Journal Classification (ASJC) codes

  • Hepatology
  • Gastroenterology

Cita questo

Conigliaro, A., Conigliaro, A., Santangelo, L., Pasquini, E., Colletti, M., Amicone, L., ... Tripodi, M. (2009). Convergence of Wnt Signaling on the HNF4α-Driven Transcription in Controlling Liver Zonation. Gastroenterology, 137, 660-672.

Convergence of Wnt Signaling on the HNF4α-Driven Transcription in Controlling Liver Zonation. / Conigliaro, Alice; Conigliaro, Alice; Santangelo, Laura; Pasquini, Emiliano; Colletti, Marta; Amicone, Laura; Cicchini, Carla; Alonzi, Tonino; Tripodi, Marco.

In: Gastroenterology, Vol. 137, 2009, pag. 660-672.

Risultato della ricerca: Article

Conigliaro, A, Conigliaro, A, Santangelo, L, Pasquini, E, Colletti, M, Amicone, L, Cicchini, C, Alonzi, T & Tripodi, M 2009, 'Convergence of Wnt Signaling on the HNF4α-Driven Transcription in Controlling Liver Zonation', Gastroenterology, vol. 137, pagg. 660-672.
Conigliaro A, Conigliaro A, Santangelo L, Pasquini E, Colletti M, Amicone L e altri. Convergence of Wnt Signaling on the HNF4α-Driven Transcription in Controlling Liver Zonation. Gastroenterology. 2009;137:660-672.
Conigliaro, Alice ; Conigliaro, Alice ; Santangelo, Laura ; Pasquini, Emiliano ; Colletti, Marta ; Amicone, Laura ; Cicchini, Carla ; Alonzi, Tonino ; Tripodi, Marco. / Convergence of Wnt Signaling on the HNF4α-Driven Transcription in Controlling Liver Zonation. In: Gastroenterology. 2009 ; Vol. 137. pagg. 660-672.
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title = "Convergence of Wnt Signaling on the HNF4{\^I}±-Driven Transcription in Controlling Liver Zonation",
abstract = "Background & Aims: In each hepatocyte, the specific repertoire of gene expression is influenced by its exact location along the portocentrovenular axis of the hepatic lobule and provides a reason for the liver functions compartmentalization defined {"}metabolic zonation.{"} So far, few molecular players controlling genetic programs of periportal (PP) and perivenular (PV) hepatocytes have been identified; the elucidation of zonation mechanisms remains a challenge for experimental hepatology. Recently, a key role in induction and maintenance of the hepatocyte heterogeneity has been ascribed to Wnt/{\^I}²-catenin pathway. We sought to clarify how this wide-ranging stimulus integrates with hepatocyte specificity. Methods: Reverse transcriptase polymerase chain reaction (RT-PCR) allowed the transcriptional profiling of hepatocytes derived from in vitro differentiation of liver stem cells. The GSK3{\^I}² inhibitor 6-bromoindirubin-3{\^a}²-oxime (BIO) was used for {\^I}²-catenin stabilization. Co-immunoprecipitations were used to study biochemical protein interactions while ChIP assays allowed the in vivo inspection of PV and PP genes regulatory regions. Results: We found that spontaneous differentiation of liver stem cells gives rise to PP hepatocytes that, after Wnt pathway activation, switch into PV hepatocytes. Next, we showed that the Wnt downstream player LEF1 interacts with the liver-enriched transcriptional factor HNF4{\^I}±. Finally, we unveiled that the BIO induced activation of PV genes correlates with LEF1 binding to both its own and HNF4{\^I}± consensus, and the repression of PP genes correlates with HNF4{\^I}± displacement from its own consensus. Conclusion: Our data show a direct and hitherto unknown convergence of the canonical Wnt signaling on the HNF4{\^I}±-driven transcription providing evidences of a mechanism controlling liver zonated gene expression. {\^A}{\circledC} 2009 AGA Institute.",
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TY - JOUR

T1 - Convergence of Wnt Signaling on the HNF4α-Driven Transcription in Controlling Liver Zonation

AU - Conigliaro, Alice

AU - Conigliaro, Alice

AU - Santangelo, Laura

AU - Pasquini, Emiliano

AU - Colletti, Marta

AU - Amicone, Laura

AU - Cicchini, Carla

AU - Alonzi, Tonino

AU - Tripodi, Marco

PY - 2009

Y1 - 2009

N2 - Background & Aims: In each hepatocyte, the specific repertoire of gene expression is influenced by its exact location along the portocentrovenular axis of the hepatic lobule and provides a reason for the liver functions compartmentalization defined "metabolic zonation." So far, few molecular players controlling genetic programs of periportal (PP) and perivenular (PV) hepatocytes have been identified; the elucidation of zonation mechanisms remains a challenge for experimental hepatology. Recently, a key role in induction and maintenance of the hepatocyte heterogeneity has been ascribed to Wnt/β-catenin pathway. We sought to clarify how this wide-ranging stimulus integrates with hepatocyte specificity. Methods: Reverse transcriptase polymerase chain reaction (RT-PCR) allowed the transcriptional profiling of hepatocytes derived from in vitro differentiation of liver stem cells. The GSK3β inhibitor 6-bromoindirubin-3â²-oxime (BIO) was used for β-catenin stabilization. Co-immunoprecipitations were used to study biochemical protein interactions while ChIP assays allowed the in vivo inspection of PV and PP genes regulatory regions. Results: We found that spontaneous differentiation of liver stem cells gives rise to PP hepatocytes that, after Wnt pathway activation, switch into PV hepatocytes. Next, we showed that the Wnt downstream player LEF1 interacts with the liver-enriched transcriptional factor HNF4α. Finally, we unveiled that the BIO induced activation of PV genes correlates with LEF1 binding to both its own and HNF4α consensus, and the repression of PP genes correlates with HNF4α displacement from its own consensus. Conclusion: Our data show a direct and hitherto unknown convergence of the canonical Wnt signaling on the HNF4α-driven transcription providing evidences of a mechanism controlling liver zonated gene expression. © 2009 AGA Institute.

AB - Background & Aims: In each hepatocyte, the specific repertoire of gene expression is influenced by its exact location along the portocentrovenular axis of the hepatic lobule and provides a reason for the liver functions compartmentalization defined "metabolic zonation." So far, few molecular players controlling genetic programs of periportal (PP) and perivenular (PV) hepatocytes have been identified; the elucidation of zonation mechanisms remains a challenge for experimental hepatology. Recently, a key role in induction and maintenance of the hepatocyte heterogeneity has been ascribed to Wnt/β-catenin pathway. We sought to clarify how this wide-ranging stimulus integrates with hepatocyte specificity. Methods: Reverse transcriptase polymerase chain reaction (RT-PCR) allowed the transcriptional profiling of hepatocytes derived from in vitro differentiation of liver stem cells. The GSK3β inhibitor 6-bromoindirubin-3â²-oxime (BIO) was used for β-catenin stabilization. Co-immunoprecipitations were used to study biochemical protein interactions while ChIP assays allowed the in vivo inspection of PV and PP genes regulatory regions. Results: We found that spontaneous differentiation of liver stem cells gives rise to PP hepatocytes that, after Wnt pathway activation, switch into PV hepatocytes. Next, we showed that the Wnt downstream player LEF1 interacts with the liver-enriched transcriptional factor HNF4α. Finally, we unveiled that the BIO induced activation of PV genes correlates with LEF1 binding to both its own and HNF4α consensus, and the repression of PP genes correlates with HNF4α displacement from its own consensus. Conclusion: Our data show a direct and hitherto unknown convergence of the canonical Wnt signaling on the HNF4α-driven transcription providing evidences of a mechanism controlling liver zonated gene expression. © 2009 AGA Institute.

KW - Animals; Blotting

KW - Cultured; Fluorescent Antibody Technique; Hepatocyte Nuclear Factor 4; Hepatocytes; Humans; Immunoprecipitation; Mice; Mice

KW - Knockout; Mice

KW - Transgenic; Reverse Transcriptase Polymerase Chain Reaction; Sensitivity and Specificity; Signal Transduction; Transfection; Wnt Proteins; beta Catenin; Gastroenterology

KW - Western; Cell Differentiation; Cell Proliferation; Cells

UR - http://hdl.handle.net/10447/241689

M3 - Article

VL - 137

SP - 660

EP - 672

JO - Gastroenterology

JF - Gastroenterology

SN - 0016-5085

ER -