TY  - JOUR
A1  - Lüth, Anja
A1  - Neuber, Corinna
A1  - Kleuser, Burkhard
T1  - Novel methods for the quantification of (2E)-hexadecenal by liquid chromatography with detection by either ESI QTOF tandem mass spectrometry or fluorescence measurement
JF  - Analytica chimica acta : an international journal devoted to all branches of analytical chemistry
N2  - Sphingosine-1-phosphate lyase (SPL) is the only known enzyme that irreversibly cleaves sphingosine-1-phosphate (S1P) into phosphoethanolamine and (2E)-hexadecenal during the final step of sphingolipid catabolism. Because S1P is involved in a wide range of physiological and diseased processes, determining the activity of the degrading enzyme is of great interest. Therefore, we developed two procedures based on liquid chromatography (LC) for analysing (2E)-hexadecenal, which is one of the two S1P degradation products. After separation, two different quantification methods were performed, tandem mass spectrometry (MS) and fluorescence detection. However, (2E)-hexadecenal as a long-chain aldehyde is not ionisable by electrospray ionisation (ESI) for MS quantification and has an insufficient number of corresponding double bonds for fluorescence detection. Therefore, we investigated 2-diphenylacetyl-1,3-indandione-1-hydrazone (DAIH) as a derivatisation reagent. DAIH transforms the aldehyde into an ionisable and fluorescent analogue for quantitative analysis. Our conditions were optimised to obtain the outstanding limit of detection (LOD) of 1 fmol per sample (30 mu L) for LC-MS/MS and 0.75 pmol per sample (200 mu l) for LC determination with fluorescence detection. We developed an extraction procedure to separate and concentrate (2E)-hexadecenal from biological samples for these measurements. To confirm our new methods, we analysed the (2E)-hexadecenal level of different cell lines and human plasma for the first time ever. Furthermore, we treated HT-29 cells with different concentrations of 4-deoxypyridoxine (DOP), which competitively inhibits pyridoxal-5-phosphate (P5P), an essential cofactor for SPL activity, and observed a significant decrease in (2E)-hexadecenal relative to the untreated cells.
KW  - (2E)-Hexadecenal
KW  - Sphingosine-1-phosphate lyase
KW  - Derivatisation
KW  - Tandem mass spectrometry
KW  - Fluorescence
Y1  - 2012
U6  - https://doi.org/10.1016/j.aca.2012.01.063
SN  - 0003-2670
VL  - 722
SP  - 70
EP  - 79
PB  - Elsevier
CY  - Amsterdam
ER  - 
TY  - JOUR
A1  - Jbeily, Nayla
A1  - Suckert, Iris
A1  - Gonnert, Falk A.
A1  - Acht, Benedikt
A1  - Bockmeyer, Clemens L.
A1  - Grossmann, Sascha D.
A1  - Blaess, Markus F.
A1  - Lüth, Anja
A1  - Deigner, Hans-Peter
A1  - Bauer, Michael
A1  - Claus, Ralf A.
T1  - Hyperresponsiveness of mice deficient in plasma-secreted sphingomyelinase reveals its pivotal role in early phase of host response
JF  - Journal of lipid research
N2  - Plasma secretion of acid sphingomyelinase is a hallmark of cellular stress response resulting in the formation of membrane embedded ceramide-enriched lipid rafts and the reorganization of receptor complexes. Consistently, decompartmentalization of ceramide formation from inert sphingomyelin has been associated with signaling events and regulation of the cellular phenotype. Herein, we addressed the question of whether the secretion of acid sphingomyelinase is involved in host response during sepsis. We found an exaggerated clinical course in mice genetically deficient in acid sphingomyelinase characterized by an increased bacterial burden, an increased phagocytotic activity, and a more pronounced cytokine storm. Moreover, on a functional level, leukocyte-endothelial interaction was found diminished in sphingomyelinase-deficient animals corresponding to a distinct leukocytes' phenotype with respect to rolling and sticking as well as expression of cellular surface proteins.(jlr) We conclude that hydrolysis of membrane-embedded sphingomyelin, triggered by circulating sphingomyelinase, plays a pivotal role in the first line of defense against invading microorganisms. This function might be essential during the early phase of infection leading to an adaptive response of remote cells and tissues.-Jbeily, N., I. Suckert, F. A. Gonnert, B. Acht, C. L. Bockmeyer, S. D. Grossmann, M. F. Blaess, A. Lueth, H.-P. Deigner, M. Bauer, and R. A. Claus. Hyperresponsiveness of mice deficient in plasma-secreted sphingomyelinase reveals its pivotal role in early phase of host response. J. Lipid Res. 2013. 54: 410-424.
KW  - sphingomyelin phosphodiesterase 1
KW  - inflammation
KW  - sepsis
KW  - gene expression
KW  - survival
KW  - leukocyte-endothelial interaction
KW  - trans-migration
KW  - organ failure
Y1  - 2013
U6  - https://doi.org/10.1194/jlr.M031625
SN  - 0022-2275
VL  - 54
IS  - 2
SP  - 410
EP  - 424
PB  - American Society for Biochemistry and Molecular Biology
CY  - Bethesda
ER  - 
TY  - JOUR
A1  - Lotinun, Sutada
A1  - Kiviranta, Riku
A1  - Matsubara, Takuma
A1  - Alzate, Jorge A.
A1  - Neff, Lynn
A1  - Lüth, Anja
A1  - Koskivirta, Ilpo
A1  - Kleuser, Burkhard
A1  - Vacher, Jean
A1  - Vuorio, Eero
A1  - Horne, William C.
A1  - Baron, Roland
T1  - Osteoclast-specific cathepsin K deletion stimulates S1P-dependent bone formation
JF  - The journal of clinical investigation
N2  - Cathepsin K (CTSK) is secreted by osteoclasts to degrade collagen and other matrix proteins during bone resorption. Global deletion of Ctsk in mice decreases bone resorption, leading to osteopetrosis, but also increases the bone formation rate (BFR). To understand how Ctsk deletion increases the BFR, we generated osteoclast- and osteoblast-targeted Ctsk knockout mice using floxed Ctsk alleles. Targeted ablation of Ctsk in hematopoietic cells, or specifically in osteoclasts and cells of the monocyte-osteoclast lineage, resulted in increased bone volume and BFR as well as osteoclast and osteoblast numbers. In contrast, targeted deletion of Ctsk in osteoblasts had no effect on bone resorption or BFR, demonstrating that the increased BFR is osteoclast dependent. Deletion of Ctsk in osteoclasts increased their sphingosine kinase 1 (Sphk1) expression. Conditioned media from Ctsk-deficient osteoclasts, which contained elevated levels of sphingosine-l-phosphate (S1P), increased alkaline phosphatase and mineralized nodules in osteoblast cultures. An S1P(1,3) receptor antagonist inhibited these responses. Osteoblasts derived from mice with Ctsk-deficient osteoclasts had an increased RANKL/OPG ratio, providing a positive feedback loop that increased the number of osteoclasts. Our data provide genetic evidence that deletion of CTSK in osteoclasts enhances bone formation in vivo by increasing the generation of osteoclast-derived S1P.
Y1  - 2013
U6  - https://doi.org/10.1172/JCI64840
SN  - 0021-9738
VL  - 123
IS  - 2
SP  - 666
EP  - 681
PB  - American Society for Clinical Investigation
CY  - Ann Arbor
ER  - 
TY  - JOUR
A1  - Böhm, Andreas
A1  - Flößer, Anja
A1  - Ermler, Swen
A1  - Fender, Anke C.
A1  - Lüth, Anja
A1  - Kleuser, Burkhard
A1  - Schrör, Karsten
A1  - Rauch, Bernhard H.
T1  - Factor-Xa-induced mitogenesis and migration require sphingosine kinase activity and S1P formation in human vascular smooth muscle cells
JF  - Cardiovascular research
N2  - Sphingosine-1-phosphate (S1P) is a cellular signalling lipid generated by sphingosine kinase-1 (SPHK1). The aim of the study was to investigate whether the activated coagulation factor-X (FXa) regulates SPHK1 transcription and the formation of S1P and subsequent mitogenesis and migration of human vascular smooth muscle cells (SMC).
 FXa induced a time- (36 h) and concentration-dependent (330 nmol/L) increase of SPHK1 mRNA and protein expression in human aortic SMC, resulting in an increased synthesis of S1P. FXa-stimulated transcription of SPHK1 was mediated by the protease-activated receptor-1 (PAR-1) and PAR-2. In human carotid artery plaques, expression of SPHK1 was observed at SMC-rich sites and was co-localized with intraplaque FX/FXa content. FXa-induced SPHK1 transcription was attenuated by inhibitors of Rho kinase (Y27632) and by protein kinase C (PKC) isoforms (GF109203X). In addition, FXa rapidly induced the activation of the small GTPase Rho A. Inhibition of signalling pathways which regulate SPHK1 expression, inhibition of its activity or siRNA-mediated SPHK1 knockdown attenuated the mitogenic and chemotactic response of human SMC to FXa.
 These data suggest that FXa induces SPHK1 expression and increases S1P formation independent of thrombin and that this involves the activation of Rho A and PKC signalling. In addition to its key function in coagulation, this direct effect of FXa on human SMC may increase cell proliferation and migration at sites of vessel injury and thereby contribute to the progression of vascular lesions.
KW  - Factor-Xa
KW  - Atherosclerosis
KW  - Proliferation
KW  - Smooth muscle cells
KW  - Sphingosine kinase-1
Y1  - 2013
U6  - https://doi.org/10.1093/cvr/cvt112
SN  - 0008-6363
VL  - 99
IS  - 3
SP  - 505
EP  - 513
PB  - Oxford Univ. Press
CY  - Oxford
ER  - 
TY  - CHAP
A1  - Natek, M.
A1  - Lüth, Anja
A1  - Kleuser, Burkhard
A1  - Schäfer-Korting, M.
A1  - Weindl, G.
T1  - CpG-oligonucleotides modulate sphingosine-1-phosphate metabolism in normal human keratinocytes
T2  - The journal of investigative dermatology
Y1  - 2012
SN  - 0022-202X
VL  - 132
IS  - 5
SP  - S112
EP  - S112
PB  - Nature Publ. Group
CY  - New York
ER  - 
TY  - JOUR
A1  - Reichel, Martin
A1  - Hoenig, Stefanie
A1  - Liebisch, Gerhard
A1  - Lüth, Anja
A1  - Kleuser, Burkhard
A1  - Gulbins, Erich
A1  - Schmitz, Gerd
A1  - Kornhuber, Johannes
T1  - Alterations of plasma glycerophospholipid and sphingolipid species in male alcohol-dependent patients
JF  - Biochimica et biophysica acta : Molecular and cell biology of lipids
N2  - Background: Alcohol abuse is a major risk factor for somatic and neuropsychiatric diseases. Despite their potential clinical importance, little is known about the alterations of plasma glycerophospholipid (GPL) and sphingolipid (SPL) species associated with alcohol abuse.
 Methods: Plasma GPL and SPL species were quantified using electrospray ionization tandem mass spectrometry in samples from 23 male alcohol-dependent patients before and after detoxification, as well as from 20 healthy male controls.
 Results: A comparison of alcohol-dependent patients with controls revealed higher phosphatidylcholine (PC; P-value = 0.008) and phosphatidylinositol (PI; P-value = 0.001) concentrations in patients before detoxification, and higher PI (P-value = 0.001) and phosphatidylethanolamine (PE)-based plasmalogen (PEP; P-value = 0.003) concentrations after detoxification. Lysophosphatidylcholines (LPC) were increased by acute intoxication (P-value = 0.002). Sphingomyelin (SM) concentration increased during detoxification (P-value = 0.011). The concentration of SM 23:0 was lower in patients (P-value = 2.79 x 10(-5)), and the concentrations of ceramide Cer d18:1/16:0 and Cer d18:1/18:0 were higher in patients (P-value = 2.45 x 10(-5) and 3.73 x 10(-5)). Activity of lysosomal acid sphingomyelinase (ASM) in patients correlated positively with the concentrations of eight LPC species, while activity of secreted ASM was inversely correlated with several PE, PI and PC species, and positively correlated with the molar ratio of PC to SM (Pearson's r = 0.432; P-value = 0.039).
 Conclusion: Plasma concentrations of numerous GPL and SPL species were altered in alcohol-dependent patients. These molecules might serve as potential biomarkers to improve the diagnosis of patients and to indicate health risks associated with alcohol abuse. Our study further indicates that there are strong interactions between plasma GPL concentrations and SPL metabolism. (C) 2015 Elsevier B.V. All rights reserved.
KW  - Acid sphingomyelinase
KW  - Alcohol dependence
KW  - Anxiety
KW  - Cardiovascular
KW  - Case-control study
KW  - Ceramide
KW  - Clinical
KW  - Depression
KW  - Diagnostic
KW  - Disease
KW  - Glycerophospholipids
KW  - Lysophosphatidylcholines
KW  - Mass spectrometry
KW  - Phosphatidylcholines
KW  - Phosphatidylinositols
KW  - Plasma
KW  - Plasmalogens
KW  - Sphingolipids
KW  - Sphingomyelin
KW  - Tandem mass spectrometry
Y1  - 2015
U6  - https://doi.org/10.1016/j.bbalip.2015.08.005
SN  - 1388-1981
SN  - 0006-3002
VL  - 1851
IS  - 11
SP  - 1501
EP  - 1510
PB  - Elsevier
CY  - Amsterdam
ER  - 
TY  - GEN
A1  - Bäumer, Wolfgang
A1  - Rossbach, Kristine
A1  - Mischke, Reinhard
A1  - Reines, Ilka
A1  - Langbein-Detsch, Ines
A1  - Lüth, Anja
A1  - Kleuser, Burkhard
T1  - Decreased concentration and enhanced metabolism of sphingosine-1-Phosphate in lesional skin of dogs with atopic dermatitis  disturbed Sphingosine-1-Phosphate homeostasis in atopic Dermatitis
T2  - The journal of investigative dermatology
Y1  - 2011
U6  - https://doi.org/10.1038/jid.2010.252
SN  - 0022-202X
VL  - 131
IS  - 1
SP  - 266
EP  - 268
PB  - Nature Publ. Group
CY  - New York
ER  - 
TY  - JOUR
A1  - Gutbier, Birgitt
A1  - Schönrock, Stefanie M.
A1  - Ehrler, Carolin
A1  - Haberberger, Rainer
A1  - Dietert, Kristina
A1  - Gruber, Achim D.
A1  - Kummer, Wolfgang
A1  - Michalick, Laura
A1  - Kuebler, Wolfgang M.
A1  - Hocke, Andreas C.
A1  - Szymanski, Kolja
A1  - Letsiou, Eleftheria
A1  - Lüth, Anja
A1  - Schumacher, Fabian
A1  - Kleuser, Burkhard
A1  - Mitchell, Timothy J.
A1  - Bertrams, Wilhelm
A1  - Schmeck, Bernd
A1  - Treue, Denise
A1  - Klauschen, Frederick
A1  - Bauer, Torsten T.
A1  - Tönnies, Mario
A1  - Weissmann, Norbert
A1  - Hippenstiel, Stefan
A1  - Suttorp, Norbert
A1  - Witzenrath, Martin
T1  - Sphingosine Kinase 1 Regulates Inflammation and Contributes to Acute Lung Injury in Pneumococcal Pneumonia via the Sphingosine-1-Phosphate Receptor 2
JF  - Critical care medicine
N2  - Objectives: Severe pneumonia may evoke acute lung injury, and sphingosine-1-phosphate is involved in the regulation of vascular permeability and immune responses. However, the role of sphingosine-1-phosphate and the sphingosine-1-phosphate producing sphingosine kinase 1 in pneumonia remains elusive. We examined the role of the sphingosine-1-phosphate system in regulating pulmonary vascular barrier function in bacterial pneumonia. Design: Controlled, in vitro, ex vivo, and in vivo laboratory study. Subjects: Female wild-type and SphK1-deficient mice, 8-10 weeks old. Human postmortem lung tissue, human blood-derived macrophages, and pulmonary microvascular endothelial cells. Interventions: Wild-type and SphK1-deficient mice were infected with Streptococcus pneumoniae. Pulmonary sphingosine-1-phosphate levels, messenger RNA expression, and permeability as well as lung morphology were analyzed. Human blood-derived macrophages and human pulmonary microvascular endothelial cells were infected with S. pneumoniae. Transcellular electrical resistance of human pulmonary microvascular endothelial cell monolayers was examined. Further, permeability of murine isolated perfused lungs was determined following exposition to sphingosine-1-phosphate and pneumolysin. Measurements and Main Results: Following S. pneumoniae infection, murine pulmonary sphingosine-1-phosphate levels and sphingosine kinase 1 and sphingosine-1-phosphate receptor 2 expression were increased. Pneumonia-induced lung hyperpermeability was reduced in SphK1(-/-) mice compared with wild-type mice. Expression of sphingosine kinase 1 in macrophages recruited to inflamed lung areas in pneumonia was observed in murine and human lungs. S. pneumoniae induced the sphingosine kinase 1/sphingosine-1-phosphate system in blood-derived macrophages and enhanced sphingosine-1-phosphate receptor 2 expression in human pulmonary microvascular endothelial cell in vitro. In isolated mouse lungs, pneumolysin-induced hyperpermeability was dose dependently and synergistically increased by sphingosine-1-phosphate. This sphingosine-1-phosphate-induced increase was reduced by inhibition of sphingosine-1-phosphate receptor 2 or its downstream effector Rho-kinase. Conclusions: Our data suggest that targeting the sphingosine kinase 1-/sphingosine-1-phosphate-/sphingosine-1-phosphate receptor 2-signaling pathway in the lung may provide a novel therapeutic perspective in pneumococcal pneumonia for prevention of acute lung injury.
KW  - acute lung injury
KW  - pneumococcal pneumonia
KW  - sphingosine kinase 1
KW  - sphingosine-1-phosphate
KW  - sphingosine-1-phosphate receptor 2
Y1  - 2018
U6  - https://doi.org/10.1097/CCM.0000000000002916
SN  - 0090-3493
SN  - 1530-0293
VL  - 46
IS  - 3
SP  - e258
EP  - e267
PB  - Lippincott Williams & Wilkins
CY  - Philadelphia
ER  -