PMID- 32882164 OWN - NLM STAT- MEDLINE DCOM- 20211108 LR - 20211208 IS - 1932-7420 (Electronic) IS - 1550-4131 (Print) IS - 1550-4131 (Linking) VI - 32 IP - 4 DP - 2020 Oct 6 TI - A Membrane-Bound Diacylglycerol Species Induces PKCϵ-Mediated Hepatic Insulin Resistance. PG - 654-664.e5 LID - S1550-4131(20)30414-9 [pii] LID - 10.1016/j.cmet.2020.08.001 [doi] AB - Nonalcoholic fatty liver disease is strongly associated with hepatic insulin resistance (HIR); however, the key lipid species and molecular mechanisms linking these conditions are widely debated. We developed a subcellular fractionation method to quantify diacylglycerol (DAG) stereoisomers and ceramides in the endoplasmic reticulum (ER), mitochondria, plasma membrane (PM), lipid droplets, and cytosol. Acute knockdown (KD) of diacylglycerol acyltransferase-2 in liver induced HIR in rats. This was due to PM sn-1,2-DAG accumulation, which promoted PKCϵ activation and insulin receptor kinase (IRK)-T1160 phosphorylation, resulting in decreased IRK-Y1162 phosphorylation. Liver PM sn-1,2-DAG content and IRK-T1160 phosphorylation were also higher in humans with HIR. In rats, liver-specific PKCϵ KD ameliorated high-fat diet-induced HIR by lowering IRK-T1160 phosphorylation, while liver-specific overexpression of constitutively active PKCϵ-induced HIR by promoting IRK-T1160 phosphorylation. These data identify PM sn-1,2-DAGs as the key pool of lipids that activate PKCϵ and that hepatic PKCϵ is both necessary and sufficient in mediating HIR. CI - Copyright (c) 2020 Elsevier Inc. All rights reserved. FAU - Lyu, Kun AU - Lyu K AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA; Department of Cellular and Molecular Physiology, Yale School of Medicine, New Haven, CT 06510, USA. FAU - Zhang, Ye AU - Zhang Y AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA; Department of Endocrinology & Metabolism, First Hospital of Jilin University, Changchun, Jilin 130021, China. FAU - Zhang, Dongyan AU - Zhang D AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA. FAU - Kahn, Mario AU - Kahn M AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA. FAU - Ter Horst, Kasper W AU - Ter Horst KW AD - Department of Endocrinology and Metabolism Amsterdam University Medical Center, 1105AZ Amsterdam, the Netherlands. FAU - Rodrigues, Marcos R S AU - Rodrigues MRS AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA; School of Medicine, State University of Ponta Grossa, Avenida General Carlos Cavalcanti, Ponta Grossa, PR 84030-900, Brazil. FAU - Gaspar, Rafael C AU - Gaspar RC AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA; Laboratory of Molecular Biology of Exercise, School of Applied Science, University of Campinas, Limeira, SP 13484-350, Brazil. FAU - Hirabara, Sandro M AU - Hirabara SM AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA; Postgraduate Interdisciplinary Program of Health Sciences, Cruzeiro do Sul University, Sao Paulo, SP 01506-000, Brazil. FAU - Luukkonen, Panu K AU - Luukkonen PK AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA. FAU - Lee, Seohyuk AU - Lee S AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA. FAU - Bhanot, Sanjay AU - Bhanot S AD - Ionis Pharmaceuticals, Carlsbad, CA 92008, USA. FAU - Rinehart, Jesse AU - Rinehart J AD - Department of Cellular and Molecular Physiology, Yale School of Medicine, New Haven, CT 06510, USA; Systems Biology Institute, Yale University, West Haven, CT 06516, USA. FAU - Blume, Niels AU - Blume N AD - CV Research, Novo Nordisk A/S, Novo Nordisk Park, 2760 Maaloev, Denmark. FAU - Rasch, Morten Gronbech AU - Rasch MG AD - Antibody Technology, Novo Nordisk A/S, Novo Nordisk Park, 2760 Maaloev, Denmark. FAU - Serlie, Mireille J AU - Serlie MJ AD - Department of Endocrinology and Metabolism Amsterdam University Medical Center, 1105AZ Amsterdam, the Netherlands. FAU - Bogan, Jonathan S AU - Bogan JS AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA; Department of Cell Biology, Yale School of Medicine, New Haven, CT 06510, USA. FAU - Cline, Gary W AU - Cline GW AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA. FAU - Samuel, Varman T AU - Samuel VT AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA; VA Connecticut Healthcare System, West Haven, CT 06516, USA. FAU - Shulman, Gerald I AU - Shulman GI AD - Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA; Department of Cellular and Molecular Physiology, Yale School of Medicine, New Haven, CT 06510, USA. Electronic address: gerald.shulman@yale.edu. LA - eng GR - UL1 TR001863/TR/NCATS NIH HHS/United States GR - R01 DK116774/DK/NIDDK NIH HHS/United States GR - R01 DK119968/DK/NIDDK NIH HHS/United States GR - P30 DK045735/DK/NIDDK NIH HHS/United States GR - P30 DK034989/DK/NIDDK NIH HHS/United States GR - R01 DK113984/DK/NIDDK NIH HHS/United States GR - R01 DK092661/DK/NIDDK NIH HHS/United States GR - R01 DK114793/DK/NIDDK NIH HHS/United States GR - I01 BX000901/BX/BLRD VA/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20200902 PL - United States TA - Cell Metab JT - Cell metabolism JID - 101233170 RN - 0 (1,2-diacylglycerol) RN - 0 (Diglycerides) RN - EC 2.7.10.1 (Receptor, Insulin) RN - EC 2.7.11.13 (Protein Kinase C-epsilon) SB - IM MH - Animals MH - Cell Membrane/*chemistry/metabolism MH - Diglycerides/chemistry/*metabolism MH - Humans MH - Insulin Resistance MH - Liver/*metabolism MH - Male MH - Phosphorylation MH - Protein Kinase C-epsilon/*metabolism MH - Rats MH - Rats, Sprague-Dawley MH - Receptor, Insulin/metabolism PMC - PMC7544641 MID - NIHMS1619429 OTO - NOTNLM OT - ceramides OT - dicylglycerols OT - hepatic glucose production OT - hepatic glycogen synthesis OT - hepatic insulin resistance OT - insulin receptor phosphorylation OT - liquid chromatography-tandem mass spectrometry OT - nonalcoholic fatty liver disease OT - protein kinase C-epsilon OT - type 2 diabetes COIS- Declaration of Interests The authors declare no competing interests. EDAT- 2020/09/04 06:00 MHDA- 2021/11/09 06:00 PMCR- 2021/10/06 CRDT- 2020/09/04 06:00 PHST- 2020/01/04 00:00 [received] PHST- 2020/06/22 00:00 [revised] PHST- 2020/08/03 00:00 [accepted] PHST- 2020/09/04 06:00 [pubmed] PHST- 2021/11/09 06:00 [medline] PHST- 2020/09/04 06:00 [entrez] PHST- 2021/10/06 00:00 [pmc-release] AID - S1550-4131(20)30414-9 [pii] AID - 10.1016/j.cmet.2020.08.001 [doi] PST - ppublish SO - Cell Metab. 2020 Oct 6;32(4):654-664.e5. doi: 10.1016/j.cmet.2020.08.001. Epub 2020 Sep 2.