GCKIII kinases control hepatocellular lipid homeostasis via shared mode of action

Metabolic dysfunction–associated steatotic liver disease has emerged as a leading global cause of chronic liver disease. Our recent translational investigations have shown that the STE20-type kinases comprising the GCKIII subfamily—MST3, STK25, and MST4—associate with hepatic lipid droplets and regu...

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Main Authors: Emmelie Cansby, Mara Caputo, Emma Andersson, Rasool Saghaleyni, Marcus Henricsson, Ying Xia, Bernice Asiedu, Matthias Blüher, L. Thomas Svensson, Andrew J. Hoy, Margit Mahlapuu
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Language:English
Published: Elsevier 2024-11-01
Series:Journal of Lipid Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S0022227524001743
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author Emmelie Cansby
Mara Caputo
Emma Andersson
Rasool Saghaleyni
Marcus Henricsson
Ying Xia
Bernice Asiedu
Matthias Blüher
L. Thomas Svensson
Andrew J. Hoy
Margit Mahlapuu
author_facet Emmelie Cansby
Mara Caputo
Emma Andersson
Rasool Saghaleyni
Marcus Henricsson
Ying Xia
Bernice Asiedu
Matthias Blüher
L. Thomas Svensson
Andrew J. Hoy
Margit Mahlapuu
author_sort Emmelie Cansby
collection DOAJ
description Metabolic dysfunction–associated steatotic liver disease has emerged as a leading global cause of chronic liver disease. Our recent translational investigations have shown that the STE20-type kinases comprising the GCKIII subfamily—MST3, STK25, and MST4—associate with hepatic lipid droplets and regulate ectopic fat storage in the liver; however, the mode of action of these proteins remains to be resolved. By comparing different combinations of the silencing of MST3, STK25, and/or MST4 in immortalized human hepatocytes, we found that their single knockdown results in a similar reduction in hepatocellular lipid content and metabolic stress, without any additive or synergistic effects observed when all three kinases are simultaneously depleted. A genome-wide yeast two-hybrid screen of the human hepatocyte library identified several interaction partners contributing to the GCKIII-mediated regulation of liver lipid homeostasis, that is, PDCD10 that protects MST3, STK25, and MST4 from degradation, MAP4K4 that regulates their activity via phosphorylation, and HSD17B11 that controls their action via a conformational change. Finally, using in vitro kinase assays on microfluidic microarrays, we pinpointed various downstream targets that are phosphorylated by the GCKIII kinases, with known functions in lipogenesis, lipolysis, and lipid secretion, as well as glucose uptake, glycolysis, hexosamine synthesis, and ubiquitination. Together, this study demonstrates that the members of the GCKIII kinase subfamily regulate hepatocyte lipid metabolism via common pathways. The results shed new light on the role of MST3, STK25, and MST4, as well as their interactions with PDCD10, MAP4K4, and HSD17B11, in the control of liver lipid homeostasis and metabolic dysfunction–associated steatotic liver disease susceptibility.
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spelling doaj-art-c9be240609c84bf68c43bf2811bc5a172025-08-20T02:06:57ZengElsevierJournal of Lipid Research0022-22752024-11-01651110066910.1016/j.jlr.2024.100669GCKIII kinases control hepatocellular lipid homeostasis via shared mode of actionEmmelie Cansby0Mara Caputo1Emma Andersson2Rasool Saghaleyni3Marcus Henricsson4Ying Xia5Bernice Asiedu6Matthias Blüher7L. Thomas Svensson8Andrew J. Hoy9Margit Mahlapuu10Department of Chemistry and Molecular Biology, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg, SwedenDepartment of Life Sciences, National Bioinformatics Infrastructure Sweden, Science for Life Laboratory, Chalmers University of Technology, Gothenburg, SwedenTranslational Science and Experimental Medicine, Research and Early Development, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden; Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Institute of Medicine, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg, SwedenHelmholtz Institute for Metabolic, Obesity, and Vascular Research (HI-MAG) of the Helmholtz Zentrum München, University of Leipzig and University Hospital Leipzig, Leipzig, GermanyDepartment of Life Sciences, National Bioinformatics Infrastructure Sweden, Science for Life Laboratory, Chalmers University of Technology, Gothenburg, SwedenSchool of Medical Sciences, Charles Perkins Centre, University of Sydney, Sydney, NSW, AustraliaDepartment of Chemistry and Molecular Biology, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg, Sweden; For correspondence: Margit MahlapuuMetabolic dysfunction–associated steatotic liver disease has emerged as a leading global cause of chronic liver disease. Our recent translational investigations have shown that the STE20-type kinases comprising the GCKIII subfamily—MST3, STK25, and MST4—associate with hepatic lipid droplets and regulate ectopic fat storage in the liver; however, the mode of action of these proteins remains to be resolved. By comparing different combinations of the silencing of MST3, STK25, and/or MST4 in immortalized human hepatocytes, we found that their single knockdown results in a similar reduction in hepatocellular lipid content and metabolic stress, without any additive or synergistic effects observed when all three kinases are simultaneously depleted. A genome-wide yeast two-hybrid screen of the human hepatocyte library identified several interaction partners contributing to the GCKIII-mediated regulation of liver lipid homeostasis, that is, PDCD10 that protects MST3, STK25, and MST4 from degradation, MAP4K4 that regulates their activity via phosphorylation, and HSD17B11 that controls their action via a conformational change. Finally, using in vitro kinase assays on microfluidic microarrays, we pinpointed various downstream targets that are phosphorylated by the GCKIII kinases, with known functions in lipogenesis, lipolysis, and lipid secretion, as well as glucose uptake, glycolysis, hexosamine synthesis, and ubiquitination. Together, this study demonstrates that the members of the GCKIII kinase subfamily regulate hepatocyte lipid metabolism via common pathways. The results shed new light on the role of MST3, STK25, and MST4, as well as their interactions with PDCD10, MAP4K4, and HSD17B11, in the control of liver lipid homeostasis and metabolic dysfunction–associated steatotic liver disease susceptibility.http://www.sciencedirect.com/science/article/pii/S0022227524001743MASLDMASHlipid dropletsliverlipotoxicitylipidomics
spellingShingle Emmelie Cansby
Mara Caputo
Emma Andersson
Rasool Saghaleyni
Marcus Henricsson
Ying Xia
Bernice Asiedu
Matthias Blüher
L. Thomas Svensson
Andrew J. Hoy
Margit Mahlapuu
GCKIII kinases control hepatocellular lipid homeostasis via shared mode of action
Journal of Lipid Research
MASLD
MASH
lipid droplets
liver
lipotoxicity
lipidomics
title GCKIII kinases control hepatocellular lipid homeostasis via shared mode of action
title_full GCKIII kinases control hepatocellular lipid homeostasis via shared mode of action
title_fullStr GCKIII kinases control hepatocellular lipid homeostasis via shared mode of action
title_full_unstemmed GCKIII kinases control hepatocellular lipid homeostasis via shared mode of action
title_short GCKIII kinases control hepatocellular lipid homeostasis via shared mode of action
title_sort gckiii kinases control hepatocellular lipid homeostasis via shared mode of action
topic MASLD
MASH
lipid droplets
liver
lipotoxicity
lipidomics
url http://www.sciencedirect.com/science/article/pii/S0022227524001743
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