Inhibition of NAE-dependent protein hyper-NEDDylation in cystic cholangiocytes halts cystogenesis in experimental models of polycystic liver disease.
Animals
Apoptosis
/ drug effects
Bile Ducts
/ pathology
Cell Proliferation
/ drug effects
Cell Survival
/ drug effects
Cyclopentanes
/ therapeutic use
Cysts
/ drug therapy
Enzyme Inhibitors
/ therapeutic use
Humans
Liver Diseases
/ drug therapy
NEDD8 Protein
/ metabolism
Protein Processing, Post-Translational
/ drug effects
Pyrimidines
/ therapeutic use
Rats
Sumoylation
Ubiquitin-Activating Enzymes
/ metabolism
Ubiquitin-Conjugating Enzymes
/ metabolism
Up-Regulation
NAE
NEDDylation
hepatic cystogenesis
posttranslational modifications
therapy
Journal
United European gastroenterology journal
ISSN: 2050-6414
Titre abrégé: United European Gastroenterol J
Pays: England
ID NLM: 101606807
Informations de publication
Date de publication:
09 2021
09 2021
Historique:
received:
03
12
2020
accepted:
01
04
2021
pubmed:
27
7
2021
medline:
1
3
2022
entrez:
26
7
2021
Statut:
ppublish
Résumé
Polycystic liver diseases (PLDs) are genetic inherited disorders characterized by the progressive growth of numerous intrahepatic biliary cysts, which are the main cause of morbidity. Previous studies revealed that cystic cholangiocytes are characterized by endoplasmic reticulum stress and aberrant posttranslational modification (PTM) of proteins, in particular hyper-SUMOylation, that promote PLD pathobiology. Protein NEDDylation is a newly characterized PTM that modulates a plethora of biological processes and its dysregulation is associated with the development and progression of several human diseases. However, the role of NEDDylation in PLD remains elusive. To explore the role of protein NEDDylation in PLD and its potential therapeutic regulatory value. Levels and functional effects of NEDDylation, including response to Pevonedistat (first-in-class selective inhibitor of the NEDDylation E1 enzyme NAE), were assessed in vitro, in vivo, and/or in patients with PLD. NEDDylated protein levels in normal and cystic human cholangiocytes were assessed by immunoprecipitation, and the proteomic profile was further analyzed by mass spectrometry. The genes involved in the NEDDylation pathway were found overexpressed (mRNA) in polycystic human and rat liver tissue, as well as in cystic cholangiocytes in culture, compared to controls. Elevated levels of NEDDylated proteins were further confirmed in cystic cholangiocytes in vitro, which diminished under Pevonedistat incubation. Pevonedistat promoted apoptotic cell death and reduced proliferation in cystic cholangiocytes in vitro. Comparative proteomic profiling of NEDD8-immunoprecipitated proteins between normal and cystic cholangiocytes in culture reported candidate proteins involved in cystogenesis, mostly associated with protein biogenesis and quality control. All these data indicate that cystic cholangiocytes display increased protein NEDDylation, contributing to cell survival and proliferation, ultimately supporting hepatic cystogenesis. Targeting of protein hyper-NEDDylation in cystic cholangiocytes inhibits cystogenesis in experimental models, representing a novel therapeutic opportunity in PLD.
Sections du résumé
BACKGROUND
Polycystic liver diseases (PLDs) are genetic inherited disorders characterized by the progressive growth of numerous intrahepatic biliary cysts, which are the main cause of morbidity. Previous studies revealed that cystic cholangiocytes are characterized by endoplasmic reticulum stress and aberrant posttranslational modification (PTM) of proteins, in particular hyper-SUMOylation, that promote PLD pathobiology. Protein NEDDylation is a newly characterized PTM that modulates a plethora of biological processes and its dysregulation is associated with the development and progression of several human diseases. However, the role of NEDDylation in PLD remains elusive.
OBJECTIVE
To explore the role of protein NEDDylation in PLD and its potential therapeutic regulatory value.
METHODS
Levels and functional effects of NEDDylation, including response to Pevonedistat (first-in-class selective inhibitor of the NEDDylation E1 enzyme NAE), were assessed in vitro, in vivo, and/or in patients with PLD. NEDDylated protein levels in normal and cystic human cholangiocytes were assessed by immunoprecipitation, and the proteomic profile was further analyzed by mass spectrometry.
RESULTS AND CONCLUSION
The genes involved in the NEDDylation pathway were found overexpressed (mRNA) in polycystic human and rat liver tissue, as well as in cystic cholangiocytes in culture, compared to controls. Elevated levels of NEDDylated proteins were further confirmed in cystic cholangiocytes in vitro, which diminished under Pevonedistat incubation. Pevonedistat promoted apoptotic cell death and reduced proliferation in cystic cholangiocytes in vitro. Comparative proteomic profiling of NEDD8-immunoprecipitated proteins between normal and cystic cholangiocytes in culture reported candidate proteins involved in cystogenesis, mostly associated with protein biogenesis and quality control. All these data indicate that cystic cholangiocytes display increased protein NEDDylation, contributing to cell survival and proliferation, ultimately supporting hepatic cystogenesis. Targeting of protein hyper-NEDDylation in cystic cholangiocytes inhibits cystogenesis in experimental models, representing a novel therapeutic opportunity in PLD.
Identifiants
pubmed: 34310849
doi: 10.1002/ueg2.12126
pmc: PMC8435261
doi:
Substances chimiques
Cyclopentanes
0
Enzyme Inhibitors
0
NEDD8 Protein
0
Pyrimidines
0
Ubiquitin-Conjugating Enzymes
EC 2.3.2.23
Ubiquitin-Activating Enzymes
EC 6.2.1.45
NAE protein, human
EC 6.3.2.-
UBE2F protein, human
EC 6.3.2.-
UBE2M protein, human
EC 6.3.2.-
pevonedistat
S3AZD8D215
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
848-859Informations de copyright
© 2021 The Authors. United European Gastroenterology Journal published by Wiley Periodicals LLC. on behalf of United European Gastroenterology.
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