Original ResearchFull Report: Basic and Translational—LiverActivation of Autophagy, Observed in Liver Tissues From Patients With Wilson Disease and From ATP7B-Deficient Animals, Protects Hepatocytes From Copper-Induced Apoptosis
Graphical abstract
Section snippets
Antibodies and Other Reagents
The full list of antibodies, small interfering RNAs (siRNAs), and primers is provided in the Supplementary Methods.
Cell Culture and Construction of Recombinant Adenoviruses
Wild-type (WT) HepG2 cells were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum, L-glutamine 2 mmol/L, and 1% penicillin and streptomycin. ATP7B-knockout (ko) HepG2 cells13 were grown in RPMI medium supplemented with 10% fetal calf serum, L-glutamine 2 mmol/L, and 1% penicillin and streptomycin. To transduce WT or ATP7B-ko cells, adenoviruses that
Loss of ATP7B and Cu Overload Trigger Autophagy in Hepatic Cells in Livers of Atp7b−/− Animals and Patients With WD
To investigate which mechanisms might be activated in ATP7B-deficient hepatocytes to protect against Cu toxicity in WD, we used an ATP7B-ko HepG2 cell line.13 Control (WT) and ATP7B-ko HepG2 cells were incubated with Cu for 24 hours and their transcriptional responses were analyzed using QuantSeq 3′ mRNA sequencing, which allows precise measurements of quantitative differences in gene expression.19 We found that Cu increased the expression levels of 1244 genes in ATP7B-ko cells vs 480 in
Discussion
The protective function of autophagy has been documented in a wide range of disorders, including conditions in which oxidative damage precipitates cell death and organismal disease.26, 27 In contrast, the role of autophagy in WD has remained obscure, although recent reports have documented activation of autophagy by Cu salts or Cu-containing anticancer compounds.10, 11, 12 In this article, we report on several novel findings demonstrating that autophagy operates as a prosurvival mechanism in WD.
Acknowledgments
The authors thank Telethon Institute of Genetics and Medicine members Cathal Wilson for critical reading of the manuscript; Andrea Ballabio, Chiara di Malta, and Gennaro Napolitano for helpful suggestions; and Angela Zampelli, Sergio Attanasio, Leandro Soria, and Edoardo Nusco for technical assistance. The authors are grateful to ITMO University/IEM members Yurii Orlov and Ekaterina Skomorokhova for help with specimen analysis. They also acknowledge support from the American and Italian Wilson
References (39)
Wilson disease
Gastroenterology
(2003)Human copper homeostasis: a network of interconnected pathways
Curr Opin Chem Biol
(2010)- et al.
High copper selectively alters lipid metabolism and cell cycle machinery in the mouse model of Wilson disease
J Biol Chem
(2007) - et al.
Overexpression of alpha-synuclein at non-toxic levels increases dopaminergic cell death induced by copper exposure via modulation of protein degradation pathways
Neurobiol Dis
(2015) - et al.
A new strain of rat for functional analysis of PINA
Brain Res Mol Brain Res
(2005) - et al.
Consequences of copper accumulation in the livers of the Atp7b−/− (Wilson disease gene) knockout mice
Am J Pathol
(2006) - et al.
Autophagy in the pathogenesis of disease
Cell
(2008) - et al.
Beclin1 controls the levels of p53 by regulating the deubiquitination activity of USP10 and USP13
Cell
(2011) - et al.
The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria
J Biol Chem
(2016) - et al.
Fulminant Wilson’s disease requiring liver transplantation in one monozygotic twin despite identical genetic mutation
Am J Transplant
(2010)
Mutations in SLC30A10 cause parkinsonism and dystonia with hypermagnesemia, polycythemia, and chronic liver disease
Am J Hum Genet
A genetic study of Wilson’s disease in the United Kingdom
Brain
Diagnosis and treatment of Wilson disease: an update
Hepatology
Mechanisms for copper acquisition, distribution and regulation
Nat Chem Biol
Urinary copper elevation in a mouse model of Wilson’s disease is a regulated process to specifically decrease the hepatic copper load
PLoS One
Dynamic internalization and recycling of a metal ion transporter: Cu homeostasis and CTR1, the human Cu(+) uptake system
J Cell Sci
Gene expression profiling of liver cells after copper overload in vivo and in vitro reveals new copper-regulated genes
J Biol Inorg Chem
Copper compound induces autophagy and apoptosis of glioma cells by reactive oxygen species and JNK activation
BMC Cancer
Activation of the MAPK11/12/13/14 (p38 MAPK) pathway regulates the transcription of autophagy genes in response to oxidative stress induced by a novel copper complex in HeLa cells
Autophagy
Cited by (154)
FGF21-mediated autophagy: Remodeling the homeostasis in response to stress in liver diseases
2024, Genes and DiseasesGinsenoside Rk1 induces autophagy-dependent apoptosis in hepatocellular carcinoma by AMPK/mTOR signaling pathway
2024, Food and Chemical ToxicologyA surge of cytosolic calcium dysregulates lysosomal function and impairs autophagy flux during cupric chloride–induced neuronal death
2024, Journal of Biological ChemistryCopper homeostasis and cuproptosis in mitochondria
2023, Life SciencesCopper-mediated novel cell death pathway in tumor cells and implications for innovative cancer therapies
2023, Biomedicine and PharmacotherapyDissecting copper biology and cancer treatment: ‘Activating Cuproptosis or suppressing Cuproplasia’
2023, Coordination Chemistry Reviews
Conflicts of interest The authors disclose no conflicts.
Funding This work was funded by Telethon, Italy (grant TIGEM-CBDM9 to Roman S. Polishchuk); AIRC, Italy (grant IG 17118 to Roman S. Polishchuk); CNR/RFBR Collaboration Program, Italy and Russia (grant 18-515-7811 to Elia Di Schiavi and Ludmila V. Puchkova); RFBR (grants 16-34-60219 and МК 2718.2018.4 to Ekaterina Y. Ilyechova); ACRI Travel Fellowship, Italy (to Ekaterina Y. Ilyechova), and in part by the Deutsche Forschungsgemeinschaft (DFG grant ZI1386/2-1 to Hans Zischka).
- §
Authors share co-senior authorship.