title: Exogenous ceramide-1-phosphate (C1P) and phospho-ceramide analogue-1 (PCERA-1) regulate key macrophage activities via distinct receptors.
abstract: Inflammation is an ensemble of tightly regulated steps, in which macrophages play an essential role. Previous reports showed that the natural sphingolipid ceramide 1-phosphate (C1P) stimulates macrophages migration, while the synthetic C1P mimic, phospho-ceramide analogue-1 (PCERA-1), suppresses production of the key pro-inflammatory cytokine TNFα and amplifies production of the key anti-inflammatory cytokine IL-10 in LPS-stimulated macrophages, via one or more unidentified G-protein coupled receptors. We show that C1P stimulated RAW264.7 macrophages migration via the NFκB pathway and MCP-1 induction, while PCERA-1 neither mimicked nor antagonized these activities. Conversely, PCERA-1 synergistically elevated LPS-dependent IL-10 expression in RAW264.7 macrophages via the cAMP-PKA-CREB signaling pathway, while C1P neither mimicked nor antagonized these activities. Interestingly, both compounds have the capacity to additively inhibit TNFα secretion; PCERA-1, but not C1P, suppressed LPS-induced TNFα expression in macrophages in a CREB-dependent manner, while C1P, but not PCERA-1, directly inhibited recombinant TNFα converting enzyme (TACE). Finally, PCERA-1 failed to interfere with binding of C1P to either the cell surface receptor or to TACE. These results thus indicate that the natural sphingolipid C1P and its synthetic analog PCERA-1 bind and activate distinct receptors expressed in RAW264.7 macrophages. Identification of these receptors will be instrumental for elucidation of novel activities of extra-cellular sphingolipids, and may pave the way for the design of new sphingolipid mimics for the treatment of inflammatory diseases, and pathologies which depend on cell migration, as in metastatic tumors.
authors.affiliation: Department of Biochemistry & Molecular Biology, Life Sciences Institute, Tel-Aviv University, Tel-Aviv 69978, Israel. Electronic address: seba120777@gmail.com.
authors.forename: Sebastián
authors.initials: S
authors.lastname: Katz
authors.affiliation: Department of Biochemistry & Molecular Biology, Life Sciences Institute, Tel-Aviv University, Tel-Aviv 69978, Israel. Electronic address: ornaerns@post.tau.ac.il.
authors.forename: Orna
authors.initials: O
authors.lastname: Ernst
authors.affiliation: Department of Biochemistry & Molecular Biology, Life Sciences Institute, Tel-Aviv University, Tel-Aviv 69978, Israel. Electronic address: davni@vcu.edu.
authors.forename: Dorit
authors.initials: D
authors.lastname: Avni
authors.affiliation: Department of Biochemistry & Molecular Biology, Life Sciences Institute, Tel-Aviv University, Tel-Aviv 69978, Israel. Electronic address: athamnam@gmail.com.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Athamna
authors.affiliation: Department of Biochemistry & Molecular Biology, Life Sciences Institute, Tel-Aviv University, Tel-Aviv 69978, Israel. Electronic address: amir.philo@gmail.com.
authors.forename: Amir
authors.initials: A
authors.lastname: Philosoph
authors.affiliation: Department of Biochemistry & Molecular Biology, University of the Basque Country, P.O. Box 644, Bilbao 48080, Spain. Electronic address: lide.arana@ehu.eus.
authors.forename: Lide
authors.initials: L
authors.lastname: Arana
authors.affiliation: Department of Biochemistry & Molecular Biology, University of the Basque Country, P.O. Box 644, Bilbao 48080, Spain. Electronic address: alber.ouro@gmail.com.
authors.forename: Alberto
authors.initials: A
authors.lastname: Ouro
authors.affiliation: Department of Biochemistry & Molecular Biology, Virginia Commonwealth University, Richmond, VA 23298-0614, United States; Hunter Holmes McGuire Veterans Administration Medical Center, Richmond, VA 23249, United States; The Massey Cancer Center, Richmond, VA 23298, United States. Electronic address: alhoeferlin@vcu.edu.
authors.forename: L Alexis
authors.initials: LA
authors.lastname: Hoeferlin
authors.affiliation: Department of Chemistry, Ben-Gurion University of the Negev, Be'er-Sheva 84105, Israel. Electronic address: meijler@bgu.ac.il.
authors.forename: Michael M
authors.initials: MM
authors.lastname: Meijler
authors.affiliation: Department of Biochemistry & Molecular Biology, Virginia Commonwealth University, Richmond, VA 23298-0614, United States; Hunter Holmes McGuire Veterans Administration Medical Center, Richmond, VA 23249, United States; The Massey Cancer Center, Richmond, VA 23298, United States. Electronic address: cechalfant@vcu.edu.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Chalfant
authors.affiliation: Department of Biochemistry & Molecular Biology, University of the Basque Country, P.O. Box 644, Bilbao 48080, Spain. Electronic address: antonio.gomez@ehu.eus.
authors.forename: Antonio
authors.initials: A
authors.lastname: Gómez-Muñoz
authors.affiliation: Department of Biochemistry & Molecular Biology, Life Sciences Institute, Tel-Aviv University, Tel-Aviv 69978, Israel. Electronic address: tsaffyz@tauex.tau.ac.il.
authors.forename: Tsaffrir
authors.initials: T
authors.lastname: Zor
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism; D000072198:ADAM17 Protein; D000818:Animals; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002518:Ceramides / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D004357:Drug Synergism; D005786:Gene Expression Regulation / Q000187:drug effects; D007249:Inflammation / Q000188:drug therapy* / Q000276:immunology; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D002518:Ceramides; D017362:Cyclic AMP Response Element-Binding Protein; D008070:Lipopolysaccharides; C541330:PCERA-1; D043562:Receptors, G-Protein-Coupled; D014409:Tumor Necrosis Factor-alpha; C065576:ceramide 1-phosphate; C539974:ceramide 1-phosphate receptor, mouse; D016753:Interleukin-10; D000242:Cyclic AMP; D051722:ADAM Proteins; D000072198:ADAM17 Protein; C000606463:Adam17 protein, mouse
keywords: C1P; IL-10; Inflammation; Macrophages; Migration; TNFα