PMID- 30452922
OWN - NLM
STAT- MEDLINE
DCOM- 20190403
LR  - 20211204
IS  - 1528-0012 (Electronic)
IS  - 0016-5085 (Linking)
VI  - 156
IP  - 4
DP  - 2019 Mar
TI  - Activation of Autophagy, Observed in Liver Tissues From Patients With Wilson 
      Disease and From ATP7B-Deficient Animals, Protects Hepatocytes From 
      Copper-Induced Apoptosis.
PG  - 1173-1189.e5
LID - S0016-5085(18)35280-6 [pii]
LID - 10.1053/j.gastro.2018.11.032 [doi]
AB  - BACKGROUND & AIMS: Wilson disease (WD) is an inherited disorder of copper 
      metabolism that leads to copper accumulation and toxicity in the liver and brain. 
      It is caused by mutations in the adenosine triphosphatase copper transporting beta 
      gene (ATP7B), which encodes a protein that transports copper from hepatocytes 
      into the bile. We studied ATP7B-deficient cells and animals to identify 
      strategies to decrease copper toxicity in patients with WD. METHODS: We used 
      RNA-seq to compare gene expression patterns between wild-type and ATP7B-knockout 
      HepG2 cells exposed to copper. We collected blood and liver tissues from 
      Atp7b(-/-) and Atp7b(+/-) (control) rats (LPP) and mice; some mice were given 5 
      daily injections of an autophagy inhibitor (spautin-1) or vehicle. We obtained 
      liver biopsies from 2 patients with WD in Italy and liver tissues from patients 
      without WD (control). Liver tissues were analyzed by immunohistochemistry, 
      immunofluorescence, cell viability, apoptosis assays, and electron and confocal 
      microscopy. Proteins were knocked down in cell lines using small interfering 
      RNAs. Levels of copper were measured in cell lysates, blood samples, liver 
      homogenates, and subcellular fractions by spectroscopy. RESULTS: After exposure 
      to copper, ATP7B-knockout cells had significant increases in the expression of 
      103 genes that regulate autophagy (including MAP1LC3A, known as LC3) compared 
      with wild-type cells. Electron and confocal microscopy visualized more autophagic 
      structures in the cytoplasm of ATP7B-knockout cells than wild-type cells after 
      copper exposure. Hepatocytes in liver tissues from patients with WD and from 
      Atp7b(-/-) mice and rats (but not controls) had multiple autophagosomes. In 
      ATP7B-knockout cells, mammalian target of rapamycin (mTOR) had decreased activity 
      and was dissociated from lysosomes; this resulted in translocation of the mTOR 
      substrate transcription factor EB to the nucleus and activation of 
      autophagy-related genes. In wild-type HepG2 cells (but not ATP7B-knockout cells), 
      exposure to copper and amino acids induced recruitment of mTOR to lysosomes. 
      Pharmacologic inhibitors of autophagy or knockdown of autophagy proteins ATG7 and 
      ATG13 induced and accelerated the death of ATP7B-knockout HepG2 cells compared 
      with wild-type cells. Autophagy protected ATP7B-knockout cells from 
      copper-induced death. CONCLUSION: ATP7B-deficient hepatocytes, such as in those 
      in patients with WD, activate autophagy in response to copper overload to prevent 
      copper-induced apoptosis. Agents designed to activate this autophagic pathway 
      might decrease copper toxicity in patients with WD.
CI  - Copyright (c) 2019 AGA Institute. Published by Elsevier Inc. All rights reserved.
FAU - Polishchuk, Elena V
AU  - Polishchuk EV
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy; 
      ITMO University, St. Petersburg, Russia; Institute of Biosciences and 
      Bioresources CNR, Italy.
FAU - Merolla, Assunta
AU  - Merolla A
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy.
FAU - Lichtmannegger, Josef
AU  - Lichtmannegger J
AD  - Institute of Molecular Toxicology and Pharmacology, Helmholtz Center Munich, 
      German Research Center for Environmental Health, Neuherberg, Germany.
FAU - Romano, Alessia
AU  - Romano A
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy.
FAU - Indrieri, Alessia
AU  - Indrieri A
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy; 
      Department of Translational Medical Science, "Federico II" University of Naples, 
      Naples, Italy.
FAU - Ilyechova, Ekaterina Y
AU  - Ilyechova EY
AD  - ITMO University, St. Petersburg, Russia; Department of Molecular Genetics, 
      Institute of Experimental Medicine, St. Petersburg, Russia.
FAU - Concilli, Mafalda
AU  - Concilli M
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy.
FAU - De Cegli, Rossella
AU  - De Cegli R
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy.
FAU - Crispino, Roberta
AU  - Crispino R
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy.
FAU - Mariniello, Marta
AU  - Mariniello M
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy.
FAU - Petruzzelli, Raffaella
AU  - Petruzzelli R
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy.
FAU - Ranucci, Giusy
AU  - Ranucci G
AD  - Division of Metabolism, IRCCS Bambino Gesu Children's Hospital, Rome, Italy.
FAU - Iorio, Raffaele
AU  - Iorio R
AD  - Department of Translational Medical Science, "Federico II" University of Naples, 
      Naples, Italy.
FAU - Pietrocola, Federico
AU  - Pietrocola F
AD  - Equipe 11 labellisee Ligue Nationale Contre le Cancer, Centre de Recherche des 
      Cordeliers, Paris, France; Institut National de la Sante et de la Recherche 
      Medicale, UMR1138, Equipe labellisee Ligue Nationale Contre le Cancer, Paris, 
      France; Universite Paris Descartes, Sorbonne Paris Cite, Paris, France; 
      Universite Pierre et Marie Curie, Paris, France; Cell Biology and Metabolomics 
      Platforms, Gustave Roussy Cancer Campus, Villejuif, France.
FAU - Einer, Claudia
AU  - Einer C
AD  - Institute of Molecular Toxicology and Pharmacology, Helmholtz Center Munich, 
      German Research Center for Environmental Health, Neuherberg, Germany.
FAU - Borchard, Sabine
AU  - Borchard S
AD  - Institute of Molecular Toxicology and Pharmacology, Helmholtz Center Munich, 
      German Research Center for Environmental Health, Neuherberg, Germany.
FAU - Zibert, Andree
AU  - Zibert A
AD  - Klinik fur Transplantationsmedizin, Universitatsklinikum Munster, Munster, 
      Germany.
FAU - Schmidt, Hartmut H
AU  - Schmidt HH
AD  - Klinik fur Transplantationsmedizin, Universitatsklinikum Munster, Munster, 
      Germany.
FAU - Di Schiavi, Elia
AU  - Di Schiavi E
AD  - Institute of Biosciences and Bioresources CNR, Italy.
FAU - Puchkova, Ludmila V
AU  - Puchkova LV
AD  - ITMO University, St. Petersburg, Russia.
FAU - Franco, Brunella
AU  - Franco B
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy; 
      Department of Translational Medical Science, "Federico II" University of Naples, 
      Naples, Italy.
FAU - Kroemer, Guido
AU  - Kroemer G
AD  - Equipe 11 labellisee Ligue Nationale Contre le Cancer, Centre de Recherche des 
      Cordeliers, Paris, France; Institut National de la Sante et de la Recherche 
      Medicale, UMR1138, Equipe labellisee Ligue Nationale Contre le Cancer, Paris, 
      France; Universite Paris Descartes, Sorbonne Paris Cite, Paris, France; 
      Universite Pierre et Marie Curie, Paris, France; Cell Biology and Metabolomics 
      Platforms, Gustave Roussy Cancer Campus, Villejuif, France; Pole de Biologie, 
      Hopital Europeen Georges Pompidou, AP-HP, Paris, France; Department of Women's 
      and Children's Health, Karolinska University Hospital, Stockholm, Sweden.
FAU - Zischka, Hans
AU  - Zischka H
AD  - Institute of Molecular Toxicology and Pharmacology, Helmholtz Center Munich, 
      German Research Center for Environmental Health, Neuherberg, Germany; Institute 
      of Toxicology and Environmental Hygiene, Technical University Munich, Munich, 
      Germany.
FAU - Polishchuk, Roman S
AU  - Polishchuk RS
AD  - Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy. 
      Electronic address: mpolish@tigem.it.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20181117
PL  - United States
TA  - Gastroenterology
JT  - Gastroenterology
JID - 0374630
RN  - 0 (Basic Helix-Loop-Helix Leucine Zipper Transcription Factors)
RN  - 0 (Benzylamines)
RN  - 0 (Quinazolines)
RN  - 0 (TFEB protein, rat)
RN  - 0 (Tcfeb protein, mouse)
RN  - 0 (spautin-1)
RN  - 789U1901C5 (Copper)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - EC 7.2.2.8 (ATP7B protein, human)
RN  - EC 7.2.2.8 (Atp7b protein, mouse)
RN  - EC 7.2.2.8 (Atp7b protein, rat)
RN  - EC 7.2.2.8 (Copper-Transporting ATPases)
SB  - IM
MH  - Animals
MH  - *Apoptosis
MH  - Autophagosomes/ultrastructure
MH  - Autophagy/drug effects/*genetics
MH  - Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism
MH  - Benzylamines/pharmacology
MH  - Cell Survival
MH  - Copper/toxicity
MH  - Copper-Transporting ATPases/*genetics/metabolism
MH  - Female
MH  - Hep G2 Cells
MH  - Hepatocytes/*physiology/ultrastructure
MH  - Hepatolenticular Degeneration/*physiopathology
MH  - Humans
MH  - Liver/*physiopathology
MH  - Male
MH  - Mice
MH  - Mice, Knockout
MH  - Microscopy, Confocal
MH  - Microscopy, Electron
MH  - Mitochondria/ultrastructure
MH  - Protein Transport
MH  - Quinazolines/pharmacology
MH  - Rats
MH  - Signal Transduction
MH  - TOR Serine-Threonine Kinases/metabolism
OTO - NOTNLM
OT  - Copper
OT  - Copper Homeostasis
OT  - Metal Toxicity
OT  - Mitophagy
EDAT- 2018/11/20 06:00
MHDA- 2019/04/04 06:00
CRDT- 2018/11/20 06:00
PHST- 2018/03/14 00:00 [received]
PHST- 2018/10/23 00:00 [revised]
PHST- 2018/11/10 00:00 [accepted]
PHST- 2018/11/20 06:00 [pubmed]
PHST- 2019/04/04 06:00 [medline]
PHST- 2018/11/20 06:00 [entrez]
AID - S0016-5085(18)35280-6 [pii]
AID - 10.1053/j.gastro.2018.11.032 [doi]
PST - ppublish
SO  - Gastroenterology. 2019 Mar;156(4):1173-1189.e5. doi: 
      10.1053/j.gastro.2018.11.032. Epub 2018 Nov 17.