PMID- 30679689 OWN - NLM STAT- MEDLINE DCOM- 20200724 LR - 20200724 IS - 2045-2322 (Electronic) IS - 2045-2322 (Linking) VI - 9 IP - 1 DP - 2019 Jan 24 TI - Role of ceramide synthase 2 in G-CSF signaling and G-CSF-R translocation into detergent-resistant membranes. PG - 747 LID - 10.1038/s41598-018-37342-8 [doi] LID - 747 AB - Ceramides are sphingolipids with defined acyl chain lengths, which are produced by corresponding ceramide synthases (CerS1-6). In experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS), the ablation of CerS2 suppresses EAE-pathology by reducing neutrophil migration into the central nervous system. This migration is induced by granulocyte-colony stimulating factor (G-CSF) signaling. G-CSF signaling leads to a signal cascade including the phosphorylation of Lyn kinase and STAT3. This in turn regulates expression of the neutrophil surface receptor chemokine receptor 2 (CXCR2) and causes translocation of the receptor into detergent-resistant membranes (DRMs). In this study we investigated the role of ceramides in G-CSF signaling. We found, that G-CSF treatment of wild type bone marrow cells (BMCs) leads to translocation of G-CSF-receptor (G-CSF-R) into DRMs. G-CSF also induces downregulation of ceramides in WT and CerS2 null BMCs, as well as upregulation of very long chain lactosylceramides. However, in CerS2 null BMCs, G-CSF failed to induce translocation of G-CSF-R into DRMs, leading to reduced phosphorylation of Lyn and reduced CXCR2 expression. Interestingly, G-CSF signaling in CerS6 null BMCs was not affected. In conclusion, very long chain ceramides are important for G-CSF signaling and translocation of G-CSF-R into DRMs. FAU - Kurz, Jennifer AU - Kurz J AD - Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Branch for Translational Medicine and Pharmacology TMP, Theodor-Stern-Kai 7, 60596, Frankfurt am Main, Germany. FAU - Barthelmes, Julia AU - Barthelmes J AD - pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University Hospital Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt/Main, Germany. FAU - Blum, Leonard AU - Blum L AD - pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University Hospital Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt/Main, Germany. FAU - Ulshofer, Thomas AU - Ulshofer T AD - Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Branch for Translational Medicine and Pharmacology TMP, Theodor-Stern-Kai 7, 60596, Frankfurt am Main, Germany. FAU - Wegner, Marthe-Susanna AU - Wegner MS AD - pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University Hospital Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt/Main, Germany. FAU - Ferreiros, Nerea AU - Ferreiros N AD - pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University Hospital Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt/Main, Germany. FAU - Roser, Luise AU - Roser L AD - Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Branch for Translational Medicine and Pharmacology TMP, Theodor-Stern-Kai 7, 60596, Frankfurt am Main, Germany. FAU - Geisslinger, Gerd AU - Geisslinger G AD - pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University Hospital Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt/Main, Germany. FAU - Grosch, Sabine AU - Grosch S AD - pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University Hospital Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt/Main, Germany. FAU - Schiffmann, Susanne AU - Schiffmann S AD - Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Branch for Translational Medicine and Pharmacology TMP, Theodor-Stern-Kai 7, 60596, Frankfurt am Main, Germany. susanne.schiffmann@ime.fraunhofer.de. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190124 PL - England TA - Sci Rep JT - Scientific reports JID - 101563288 RN - 0 (Detergents) RN - 0 (Lactosylceramides) RN - 0 (Receptors, Granulocyte Colony-Stimulating Factor) RN - 0 (STAT3 Transcription Factor) RN - 143011-72-7 (Granulocyte Colony-Stimulating Factor) RN - EC 2.3.1.24 (CERS6 protein, mouse) RN - EC 2.3.1.24 (Cers2 protein, mouse) RN - EC 2.3.1.24 (Sphingosine N-Acyltransferase) RN - EC 2.7.10.2 (lyn protein-tyrosine kinase) RN - EC 2.7.10.2 (src-Family Kinases) SB - IM MH - Animals MH - Bone Marrow Cells/drug effects/metabolism MH - Cell Membrane/drug effects/genetics MH - Cell Movement/drug effects MH - Detergents/pharmacology MH - Disease Models, Animal MH - Encephalomyelitis, Autoimmune, Experimental/*genetics/metabolism/pathology MH - Gene Expression Regulation/drug effects MH - Granulocyte Colony-Stimulating Factor/*genetics MH - Humans MH - Lactosylceramides/metabolism MH - Mice MH - Multiple Sclerosis/*genetics/metabolism/pathology MH - Neutrophils/drug effects MH - Protein Transport/drug effects MH - Receptors, Granulocyte Colony-Stimulating Factor/*genetics MH - STAT3 Transcription Factor/genetics MH - Sphingosine N-Acyltransferase/*genetics MH - src-Family Kinases/genetics PMC - PMC6345911 COIS- The authors declare no competing interests. EDAT- 2019/01/27 06:00 MHDA- 2020/07/25 06:00 PMCR- 2019/01/24 CRDT- 2019/01/26 06:00 PHST- 2018/07/24 00:00 [received] PHST- 2018/12/06 00:00 [accepted] PHST- 2019/01/26 06:00 [entrez] PHST- 2019/01/27 06:00 [pubmed] PHST- 2020/07/25 06:00 [medline] PHST- 2019/01/24 00:00 [pmc-release] AID - 10.1038/s41598-018-37342-8 [pii] AID - 37342 [pii] AID - 10.1038/s41598-018-37342-8 [doi] PST - epublish SO - Sci Rep. 2019 Jan 24;9(1):747. doi: 10.1038/s41598-018-37342-8.