Cargando…

Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue

Fenretinide (FEN) is a synthetic retinoid that inhibits obesity and insulin resistance in high-fat diet (HFD)-fed mice and completely prevents 3T3-L1 pre-adipocyte differentiation. The aim of this study was to determine the mechanism(s) of FEN action in 3T3-L1 adipocytes and in mice. We used the 3T3...

Descripción completa

Detalles Bibliográficos
Autores principales: Mcilroy, George D., Tammireddy, Seshu R., Maskrey, Benjamin H., Grant, Louise, Doherty, Mary K., Watson, David G., Delibegović, Mirela, Whitfield, Phillip D., Mody, Nimesh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4762576/
https://www.ncbi.nlm.nih.gov/pubmed/26592777
http://dx.doi.org/10.1016/j.bcp.2015.11.017
_version_ 1782417131353145344
author Mcilroy, George D.
Tammireddy, Seshu R.
Maskrey, Benjamin H.
Grant, Louise
Doherty, Mary K.
Watson, David G.
Delibegović, Mirela
Whitfield, Phillip D.
Mody, Nimesh
author_facet Mcilroy, George D.
Tammireddy, Seshu R.
Maskrey, Benjamin H.
Grant, Louise
Doherty, Mary K.
Watson, David G.
Delibegović, Mirela
Whitfield, Phillip D.
Mody, Nimesh
author_sort Mcilroy, George D.
collection PubMed
description Fenretinide (FEN) is a synthetic retinoid that inhibits obesity and insulin resistance in high-fat diet (HFD)-fed mice and completely prevents 3T3-L1 pre-adipocyte differentiation. The aim of this study was to determine the mechanism(s) of FEN action in 3T3-L1 adipocytes and in mice. We used the 3T3-L1 model of adipogenesis, fully differentiated 3T3-L1 adipocytes and adipose tissue from HFD-induced obese mice to investigate the mechanisms of FEN action. We measured expression of adipogenic and retinoid genes by qPCR and activation of nutrient-signalling pathways by western blotting. Global lipid and metabolite analysis was performed and specific ceramide lipid species measured by liquid chromatography-mass spectrometry. We provide direct evidence that FEN inhibits 3T3-L1 adipogenesis via RA-receptor (RAR)-dependent signaling. However, RARα antagonism did not prevent FEN-induced decreases in lipid levels in mature 3T3-L1 adipocytes, suggesting an RAR-independent mechanism. Lipidomics analysis revealed that FEN increased dihydroceramide lipid species 5- to 16-fold in adipocytes, indicating an inhibition of the final step of ceramide biosynthesis. A similar blockade in adipose tissue from FEN-treated obese mice was associated with a complete normalisation of impaired mitochondrial β-oxidation and tricarboxylic acid cycle flux. The FEN catabolite, 4-oxo-N-(4-hydroxyphenyl)retinamide (4-OXO), also decreased lipid accumulation without affecting adipogenesis. FEN and 4-OXO (but not RA) treatment additionally led to the activation of p38-MAPK, peIF2α and autophagy markers in adipocytes. Overall our data reveals FEN utilises both RAR-dependent and -independent pathways to regulate adipocyte biology, both of which may be required for FEN to prevent obesity and insulin resistance in vivo.
format Online
Article
Text
id pubmed-4762576
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Elsevier Science
record_format MEDLINE/PubMed
spelling pubmed-47625762016-03-07 Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue Mcilroy, George D. Tammireddy, Seshu R. Maskrey, Benjamin H. Grant, Louise Doherty, Mary K. Watson, David G. Delibegović, Mirela Whitfield, Phillip D. Mody, Nimesh Biochem Pharmacol Article Fenretinide (FEN) is a synthetic retinoid that inhibits obesity and insulin resistance in high-fat diet (HFD)-fed mice and completely prevents 3T3-L1 pre-adipocyte differentiation. The aim of this study was to determine the mechanism(s) of FEN action in 3T3-L1 adipocytes and in mice. We used the 3T3-L1 model of adipogenesis, fully differentiated 3T3-L1 adipocytes and adipose tissue from HFD-induced obese mice to investigate the mechanisms of FEN action. We measured expression of adipogenic and retinoid genes by qPCR and activation of nutrient-signalling pathways by western blotting. Global lipid and metabolite analysis was performed and specific ceramide lipid species measured by liquid chromatography-mass spectrometry. We provide direct evidence that FEN inhibits 3T3-L1 adipogenesis via RA-receptor (RAR)-dependent signaling. However, RARα antagonism did not prevent FEN-induced decreases in lipid levels in mature 3T3-L1 adipocytes, suggesting an RAR-independent mechanism. Lipidomics analysis revealed that FEN increased dihydroceramide lipid species 5- to 16-fold in adipocytes, indicating an inhibition of the final step of ceramide biosynthesis. A similar blockade in adipose tissue from FEN-treated obese mice was associated with a complete normalisation of impaired mitochondrial β-oxidation and tricarboxylic acid cycle flux. The FEN catabolite, 4-oxo-N-(4-hydroxyphenyl)retinamide (4-OXO), also decreased lipid accumulation without affecting adipogenesis. FEN and 4-OXO (but not RA) treatment additionally led to the activation of p38-MAPK, peIF2α and autophagy markers in adipocytes. Overall our data reveals FEN utilises both RAR-dependent and -independent pathways to regulate adipocyte biology, both of which may be required for FEN to prevent obesity and insulin resistance in vivo. Elsevier Science 2016-01-15 /pmc/articles/PMC4762576/ /pubmed/26592777 http://dx.doi.org/10.1016/j.bcp.2015.11.017 Text en © 2015 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Mcilroy, George D.
Tammireddy, Seshu R.
Maskrey, Benjamin H.
Grant, Louise
Doherty, Mary K.
Watson, David G.
Delibegović, Mirela
Whitfield, Phillip D.
Mody, Nimesh
Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue
title Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue
title_full Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue
title_fullStr Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue
title_full_unstemmed Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue
title_short Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue
title_sort fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4762576/
https://www.ncbi.nlm.nih.gov/pubmed/26592777
http://dx.doi.org/10.1016/j.bcp.2015.11.017
work_keys_str_mv AT mcilroygeorged fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue
AT tammireddyseshur fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue
AT maskreybenjaminh fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue
AT grantlouise fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue
AT dohertymaryk fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue
AT watsondavidg fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue
AT delibegovicmirela fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue
AT whitfieldphillipd fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue
AT modynimesh fenretinidemediatedretinoicacidreceptorsignallingandinhibitionofceramidebiosynthesisregulatesadipogenesislipidaccumulationmitochondrialfunctionandnutrientstresssignallinginadipocytesandadiposetissue