ASMase regulates autophagy and lysosomal membrane permeabilization and its inhibition prevents early stage non-alcoholic steatohepatitis

Raquel Fucho, Laura Martínez, Anna Baulies, Sandra Torres, Nuria Tarrats, Anna Fernandez, Vicente Ribas, Alma M. Astudillo, Jesús Balsinde, Pablo Garcia-Rovés, Montserrat Elena, Ina Bergheim, Sophie Lotersztajn, Christian Trautwein, Hanna Appelqvist, Adrienne W. Paton, James C. Paton, Mark J. Czaja, Neil Kaplowitz, Jose C. Fernandez-ChecaCarmen García-Ruiz

Research output: Contribution to journalArticle

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Abstract

Background & Aims: Acid sphingomyelinase (ASMase) is activated in non-alcoholic steatohepatitis (NASH). However, the contribution of ASMase to NASH is poorly understood and limited to hepatic steatosis and glucose metabolism. Here we examined the role of ASMase in high fat diet (HFD)-induced NASH. Methods: Autophagy, endoplasmic reticulum (ER) stress and lysosomal membrane permeabilization (LMP) were determined in ASMase<sup>-/-</sup> mice fed a HFD. The impact of pharmacological ASMase inhibition on NASH was analyzed in wild type mice fed a HFD. Results: ASMase deficiency determined resistance to hepatic steatosis mediated by a HFD or methionine-choline deficient diet. ASMase<sup>-/-</sup> mice were resistant to HFD-induced hepatic ER stress, but sensitive to tunicamycin-mediated ER stress, indicating selectivity in the resistance of ASMase<sup>-/-</sup> mice to ER stress and steatosis. Autophagic flux, determined in the presence of rapamycin and/or chloroquine, was lower in primary mouse hepatocytes (PMH) from ASMase<sup>-/-</sup> mice and accompanied by increased p62 levels, suggesting autophagic impairment. Moreover, autophagy suppression by chloroquine and brefeldin A caused ER stress in PMH from ASMase<sup>+/+</sup> mice but not in ASMase<sup>-/-</sup> mice. ASMase<sup>-/-</sup> PMH exhibited increased lysosomal cholesterol loading, decreased LMP and apoptosis resistance induced by O-methylserine dodecylamide hydrochloride or palmitic acid, effects that were reversed by decreasing cholesterol levels by oxysterol 25- hydroxycholesterol. In vivo pharmacological ASMase inhibition by amitriptyline, a widely used tricyclic antidepressant, protected wild type mice against HFD-induced hepatic steatosis, fibrosis, and liver damage, effects indicative of early-stage NASH. Conclusions: These findings underscore a critical role for ASMase in diet-induced NASH and suggest the potential of amitriptyline as a treatment for patients with NASH.

Original languageEnglish (US)
Pages (from-to)1126-1134
Number of pages9
JournalJournal of Hepatology
Volume61
Issue number5
DOIs
StatePublished - 2014

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Sphingomyelin Phosphodiesterase
Autophagy
Fatty Liver
Acids
Membranes
High Fat Diet
Endoplasmic Reticulum Stress
Hepatocytes
Amitriptyline
Liver
Chloroquine
Type A Niemann-Pick Disease
Cholesterol
Pharmacology
Diet
Brefeldin A
Tunicamycin
Palmitic Acid
Tricyclic Antidepressive Agents
Sirolimus

Keywords

  • Autophagy
  • Ceramide
  • Endoplasmic reticulum stress
  • Fatty liver
  • Lysosomal membrane permeabilization

ASJC Scopus subject areas

  • Hepatology

Cite this

ASMase regulates autophagy and lysosomal membrane permeabilization and its inhibition prevents early stage non-alcoholic steatohepatitis. / Fucho, Raquel; Martínez, Laura; Baulies, Anna; Torres, Sandra; Tarrats, Nuria; Fernandez, Anna; Ribas, Vicente; Astudillo, Alma M.; Balsinde, Jesús; Garcia-Rovés, Pablo; Elena, Montserrat; Bergheim, Ina; Lotersztajn, Sophie; Trautwein, Christian; Appelqvist, Hanna; Paton, Adrienne W.; Paton, James C.; Czaja, Mark J.; Kaplowitz, Neil; Fernandez-Checa, Jose C.; García-Ruiz, Carmen.

In: Journal of Hepatology, Vol. 61, No. 5, 2014, p. 1126-1134.

Research output: Contribution to journalArticle

Fucho, R, Martínez, L, Baulies, A, Torres, S, Tarrats, N, Fernandez, A, Ribas, V, Astudillo, AM, Balsinde, J, Garcia-Rovés, P, Elena, M, Bergheim, I, Lotersztajn, S, Trautwein, C, Appelqvist, H, Paton, AW, Paton, JC, Czaja, MJ, Kaplowitz, N, Fernandez-Checa, JC & García-Ruiz, C 2014, 'ASMase regulates autophagy and lysosomal membrane permeabilization and its inhibition prevents early stage non-alcoholic steatohepatitis', Journal of Hepatology, vol. 61, no. 5, pp. 1126-1134. https://doi.org/10.1016/j.jhep.2014.06.009
Fucho, Raquel ; Martínez, Laura ; Baulies, Anna ; Torres, Sandra ; Tarrats, Nuria ; Fernandez, Anna ; Ribas, Vicente ; Astudillo, Alma M. ; Balsinde, Jesús ; Garcia-Rovés, Pablo ; Elena, Montserrat ; Bergheim, Ina ; Lotersztajn, Sophie ; Trautwein, Christian ; Appelqvist, Hanna ; Paton, Adrienne W. ; Paton, James C. ; Czaja, Mark J. ; Kaplowitz, Neil ; Fernandez-Checa, Jose C. ; García-Ruiz, Carmen. / ASMase regulates autophagy and lysosomal membrane permeabilization and its inhibition prevents early stage non-alcoholic steatohepatitis. In: Journal of Hepatology. 2014 ; Vol. 61, No. 5. pp. 1126-1134.
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abstract = "Background & Aims: Acid sphingomyelinase (ASMase) is activated in non-alcoholic steatohepatitis (NASH). However, the contribution of ASMase to NASH is poorly understood and limited to hepatic steatosis and glucose metabolism. Here we examined the role of ASMase in high fat diet (HFD)-induced NASH. Methods: Autophagy, endoplasmic reticulum (ER) stress and lysosomal membrane permeabilization (LMP) were determined in ASMase-/- mice fed a HFD. The impact of pharmacological ASMase inhibition on NASH was analyzed in wild type mice fed a HFD. Results: ASMase deficiency determined resistance to hepatic steatosis mediated by a HFD or methionine-choline deficient diet. ASMase-/- mice were resistant to HFD-induced hepatic ER stress, but sensitive to tunicamycin-mediated ER stress, indicating selectivity in the resistance of ASMase-/- mice to ER stress and steatosis. Autophagic flux, determined in the presence of rapamycin and/or chloroquine, was lower in primary mouse hepatocytes (PMH) from ASMase-/- mice and accompanied by increased p62 levels, suggesting autophagic impairment. Moreover, autophagy suppression by chloroquine and brefeldin A caused ER stress in PMH from ASMase+/+ mice but not in ASMase-/- mice. ASMase-/- PMH exhibited increased lysosomal cholesterol loading, decreased LMP and apoptosis resistance induced by O-methylserine dodecylamide hydrochloride or palmitic acid, effects that were reversed by decreasing cholesterol levels by oxysterol 25- hydroxycholesterol. In vivo pharmacological ASMase inhibition by amitriptyline, a widely used tricyclic antidepressant, protected wild type mice against HFD-induced hepatic steatosis, fibrosis, and liver damage, effects indicative of early-stage NASH. Conclusions: These findings underscore a critical role for ASMase in diet-induced NASH and suggest the potential of amitriptyline as a treatment for patients with NASH.",
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T1 - ASMase regulates autophagy and lysosomal membrane permeabilization and its inhibition prevents early stage non-alcoholic steatohepatitis

AU - Fucho, Raquel

AU - Martínez, Laura

AU - Baulies, Anna

AU - Torres, Sandra

AU - Tarrats, Nuria

AU - Fernandez, Anna

AU - Ribas, Vicente

AU - Astudillo, Alma M.

AU - Balsinde, Jesús

AU - Garcia-Rovés, Pablo

AU - Elena, Montserrat

AU - Bergheim, Ina

AU - Lotersztajn, Sophie

AU - Trautwein, Christian

AU - Appelqvist, Hanna

AU - Paton, Adrienne W.

AU - Paton, James C.

AU - Czaja, Mark J.

AU - Kaplowitz, Neil

AU - Fernandez-Checa, Jose C.

AU - García-Ruiz, Carmen

PY - 2014

Y1 - 2014

N2 - Background & Aims: Acid sphingomyelinase (ASMase) is activated in non-alcoholic steatohepatitis (NASH). However, the contribution of ASMase to NASH is poorly understood and limited to hepatic steatosis and glucose metabolism. Here we examined the role of ASMase in high fat diet (HFD)-induced NASH. Methods: Autophagy, endoplasmic reticulum (ER) stress and lysosomal membrane permeabilization (LMP) were determined in ASMase-/- mice fed a HFD. The impact of pharmacological ASMase inhibition on NASH was analyzed in wild type mice fed a HFD. Results: ASMase deficiency determined resistance to hepatic steatosis mediated by a HFD or methionine-choline deficient diet. ASMase-/- mice were resistant to HFD-induced hepatic ER stress, but sensitive to tunicamycin-mediated ER stress, indicating selectivity in the resistance of ASMase-/- mice to ER stress and steatosis. Autophagic flux, determined in the presence of rapamycin and/or chloroquine, was lower in primary mouse hepatocytes (PMH) from ASMase-/- mice and accompanied by increased p62 levels, suggesting autophagic impairment. Moreover, autophagy suppression by chloroquine and brefeldin A caused ER stress in PMH from ASMase+/+ mice but not in ASMase-/- mice. ASMase-/- PMH exhibited increased lysosomal cholesterol loading, decreased LMP and apoptosis resistance induced by O-methylserine dodecylamide hydrochloride or palmitic acid, effects that were reversed by decreasing cholesterol levels by oxysterol 25- hydroxycholesterol. In vivo pharmacological ASMase inhibition by amitriptyline, a widely used tricyclic antidepressant, protected wild type mice against HFD-induced hepatic steatosis, fibrosis, and liver damage, effects indicative of early-stage NASH. Conclusions: These findings underscore a critical role for ASMase in diet-induced NASH and suggest the potential of amitriptyline as a treatment for patients with NASH.

AB - Background & Aims: Acid sphingomyelinase (ASMase) is activated in non-alcoholic steatohepatitis (NASH). However, the contribution of ASMase to NASH is poorly understood and limited to hepatic steatosis and glucose metabolism. Here we examined the role of ASMase in high fat diet (HFD)-induced NASH. Methods: Autophagy, endoplasmic reticulum (ER) stress and lysosomal membrane permeabilization (LMP) were determined in ASMase-/- mice fed a HFD. The impact of pharmacological ASMase inhibition on NASH was analyzed in wild type mice fed a HFD. Results: ASMase deficiency determined resistance to hepatic steatosis mediated by a HFD or methionine-choline deficient diet. ASMase-/- mice were resistant to HFD-induced hepatic ER stress, but sensitive to tunicamycin-mediated ER stress, indicating selectivity in the resistance of ASMase-/- mice to ER stress and steatosis. Autophagic flux, determined in the presence of rapamycin and/or chloroquine, was lower in primary mouse hepatocytes (PMH) from ASMase-/- mice and accompanied by increased p62 levels, suggesting autophagic impairment. Moreover, autophagy suppression by chloroquine and brefeldin A caused ER stress in PMH from ASMase+/+ mice but not in ASMase-/- mice. ASMase-/- PMH exhibited increased lysosomal cholesterol loading, decreased LMP and apoptosis resistance induced by O-methylserine dodecylamide hydrochloride or palmitic acid, effects that were reversed by decreasing cholesterol levels by oxysterol 25- hydroxycholesterol. In vivo pharmacological ASMase inhibition by amitriptyline, a widely used tricyclic antidepressant, protected wild type mice against HFD-induced hepatic steatosis, fibrosis, and liver damage, effects indicative of early-stage NASH. Conclusions: These findings underscore a critical role for ASMase in diet-induced NASH and suggest the potential of amitriptyline as a treatment for patients with NASH.

KW - Autophagy

KW - Ceramide

KW - Endoplasmic reticulum stress

KW - Fatty liver

KW - Lysosomal membrane permeabilization

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