PMID- 30655217 OWN - NLM STAT- MEDLINE DCOM- 20191211 LR - 20231006 IS - 2212-8778 (Electronic) IS - 2212-8778 (Linking) VI - 21 DP - 2019 Mar TI - The role of C16:0 ceramide in the development of obesity and type 2 diabetes: CerS6 inhibition as a novel therapeutic approach. PG - 36-50 LID - S2212-8778(18)30805-6 [pii] LID - 10.1016/j.molmet.2018.12.008 [doi] AB - OBJECTIVE: Ectopic fat deposition is associated with increased tissue production of ceramides. Recent genetic mouse studies suggest that specific sphingolipid C16:0 ceramide produced by ceramide synthase 6 (CerS6) plays an important role in the development of insulin resistance. However, the therapeutic potential of CerS6 inhibition not been demonstrated. Therefore, we pharmacologically investigated the selective ablation of CerS6 using antisense oligonucleotides (ASO) in obese insulin resistance animal models. METHODS: We utilized ASO as therapeutic modality, CerS6 ASO molecules designed and synthesized were initially screened for in-vitro knock-down (KD) potency and cytotoxicity. ASOs with >85% inhibition of CerS6 mRNA were selected for further investigations. Most promising ASOs verified for in-vivo KD efficacy in healthy mice. CerS6 ASO (AAGATGAGCCGCACC) was found most active with hepatic reduction of CerS6 mRNA expression. Prior to longitudinal metabolic studies, we performed a dose titration target engagement analysis with CerS6 ASO in healthy mice to select the optimal dose. Next, we utilized leptin deficiency ob/ob and high fat diet (HFD) induced obese mouse models for pharmacological efficacy study. RESULTS: CerS6 expression were significantly elevated in the liver and brown adipose, this was correlated with significantly elevated C16:0 ceramide concentrations in plasma and liver. Treatment with CerS6 ASO selectively reduced CerS6 expression by approximately 90% predominantly in the liver and this CerS6 KD resulted in a significant reduction of C16:0 ceramide by about 50% in both liver and plasma. CerS6 KD resulted in lower body weight gain and accompanied by a significant reduction in whole body fat and fed/fasted blood glucose levels (1% reduction in HbA1c). Moreover, ASO-mediated CerS6 KD significantly improved oral glucose tolerance (during oGTT) and mice displayed improved insulin sensitivity. Thus, CerS6 appear to play an important role in the development of obesity and insulin resistance. CONCLUSIONS: Our investigations identified specific and selective therapeutic valid ASO for CerS6 ablation in in-vivo. CerS6 should specifically be targeted for the reduction of C16:0 ceramides, that results in amelioration of insulin resistance, hyperglycemia and obesity. CerS6 mediated C16:0 ceramide reduction could be a potentially attractive target for the treatment of insulin resistance, obesity and type 2 diabetes. CI - Copyright (c) 2019 The Authors. Published by Elsevier GmbH.. All rights reserved. FAU - Raichur, Suryaprakash AU - Raichur S AD - Evotec International GmbH, Marie-Curie-Strasse 7, 37079 Goettingen, Germany. Electronic address: Surya.Prakash@evotec.com. FAU - Brunner, Bodo AU - Brunner B AD - Sanofi-Aventis Deutschland GmbH, TA Diabetes, Industriepark Hochst, D-65926 Frankfurt am Main, Germany. FAU - Bielohuby, Maximilian AU - Bielohuby M AD - Sanofi-Aventis Deutschland GmbH, TA Diabetes, Industriepark Hochst, D-65926 Frankfurt am Main, Germany. FAU - Hansen, Gitte AU - Hansen G AD - Gubra ApS, Horsholm Kongevej 11B, 2970 Horsholm, Denmark. FAU - Pfenninger, Anja AU - Pfenninger A AD - Sanofi-Aventis Deutschland GmbH, TA Diabetes, Industriepark Hochst, D-65926 Frankfurt am Main, Germany. FAU - Wang, Bing AU - Wang B AD - Analytical Research & Development US Predevelopment Sciences, Sanofi, 153 Second Avenue, Waltham, MA 02451, USA. FAU - Bruning, Jens C AU - Bruning JC AD - Max Planck Institute for Metabolic Research, Gleueler Str. 50, D-50931 Cologne, Germany. FAU - Larsen, Philip Just AU - Larsen PJ AD - Grunenthal GmbH, 52072 Aachen, Germany. FAU - Tennagels, Norbert AU - Tennagels N AD - Sanofi-Aventis Deutschland GmbH, TA Diabetes, Industriepark Hochst, D-65926 Frankfurt am Main, Germany. Electronic address: Norbert.Tennagels@sanofi.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190102 PL - Germany TA - Mol Metab JT - Molecular metabolism JID - 101605730 RN - 0 (Blood Glucose) RN - 0 (Ceramides) RN - 0 (Leptin) RN - 0 (Membrane Proteins) RN - 0 (Oligonucleotides, Antisense) RN - 0 (Thionucleotides) RN - 4201-58-5 (N-palmitoylsphingosine) RN - EC 2.3.1.24 (CERS6 protein, human) RN - EC 2.3.1.24 (CERS6 protein, mouse) RN - EC 2.3.1.24 (Sphingosine N-Acyltransferase) SB - IM MH - Adipose Tissue, Brown/metabolism MH - Animals MH - Blood Glucose/metabolism MH - Ceramides/*metabolism MH - Diabetes Mellitus, Type 2/*metabolism MH - Diet, High-Fat/adverse effects MH - Disease Models, Animal MH - Gene Knockdown Techniques MH - Hep G2 Cells MH - Humans MH - Insulin Resistance MH - Leptin/deficiency MH - Liver/metabolism MH - Male MH - Membrane Proteins/antagonists & inhibitors/genetics/metabolism MH - Mice MH - Mice, Inbred C57BL MH - Mice, Obese MH - Obesity/etiology/*metabolism MH - Oligonucleotides, Antisense/*metabolism/pharmacology MH - Sphingosine N-Acyltransferase/antagonists & inhibitors/genetics/*metabolism MH - Thionucleotides MH - Weight Gain PMC - PMC6407366 OTO - NOTNLM OT - Antisense oligonucleotide OT - Ceramides OT - Insulin resistance OT - Obesity OT - Sphingolipids OT - Type 2 diabetes EDAT- 2019/01/19 06:00 MHDA- 2019/12/18 06:00 PMCR- 2019/01/02 CRDT- 2019/01/19 06:00 PHST- 2018/08/10 00:00 [received] PHST- 2018/12/20 00:00 [revised] PHST- 2018/12/21 00:00 [accepted] PHST- 2019/01/19 06:00 [pubmed] PHST- 2019/12/18 06:00 [medline] PHST- 2019/01/19 06:00 [entrez] PHST- 2019/01/02 00:00 [pmc-release] AID - S2212-8778(18)30805-6 [pii] AID - 10.1016/j.molmet.2018.12.008 [doi] PST - ppublish SO - Mol Metab. 2019 Mar;21:36-50. doi: 10.1016/j.molmet.2018.12.008. Epub 2019 Jan 2.