title: Sphingolipids Contribute to Human Atherosclerotic Plaque Inflammation.
abstract: Lipids are central to the development of atherosclerotic plaques. Specifically, which lipids are culprits remains controversial, and promising targets have failed in clinical studies. Sphingolipids are bioactive lipids present in atherosclerotic plaques, and they have been suggested to have both proatherogenic and antiatherogenic. However, the biological effects of these lipids remain unknown in the human atherosclerotic plaque. The aim of this study was to assess plaque levels of sphingolipids and investigate their potential association with and contribution to plaque vulnerability.
  
  
  
  Glucosylceramide, lactosylceramide, ceramide, dihydroceramide, sphingomyelin, and sphingosine-1-phosphate were analyzed in homogenates from 200 human carotid plaques using mass spectrometry. Inflammatory activity was determined by analyzing plaque levels of cytokines and plaque histology. Caspase-3 was analyzed by ELISA technique. Expression of regulatory enzymes was analyzed with RNA sequencing. Human coronary artery smooth muscle cells were used to analyze the potential role of the 6 sphingolipids as inducers of plaque inflammation and cellular apoptosis in vitro. All sphingolipids were increased in plaques associated with symptoms and correlated with inflammatory cytokines. All sphingolipids, except sphingosine-1-phosphate, also correlated with histological markers of plaque instability. Lactosylceramide, ceramide, sphingomyelin, and sphingosine-1-phosphate correlated with caspase-3 activity. In vitro experiments revealed that glucosylceramide, lactosylceramide, and ceramide induced cellular apoptosis. All analyzed sphingolipids induced an inflammatory response in human coronary artery smooth muscle cells.
  
  
  
  This study shows for the first time that sphingolipids and particularly glucosylceramide are associated with and are possible inducers of plaque inflammation and instability, pointing to sphingolipid metabolic pathways as possible novel therapeutic targets.
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.). Andreas.Edsfeldt@med.lu.se.
authors.forename: Andreas
authors.initials: A
authors.lastname: Edsfeldt
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Pontus
authors.initials: P
authors.lastname: Dunér
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Marcus
authors.initials: M
authors.lastname: Ståhlman
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Ines G
authors.initials: IG
authors.lastname: Mollet
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Asciutto
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Helena
authors.initials: H
authors.lastname: Grufman
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Mihaela
authors.initials: M
authors.lastname: Nitulescu
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Ana Flor
authors.initials: AF
authors.lastname: Persson
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Fisher
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Olle
authors.initials: O
authors.lastname: Melander
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Marju
authors.initials: M
authors.lastname: Orho-Melander
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Jan
authors.initials: J
authors.lastname: Borén
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Jan
authors.initials: J
authors.lastname: Nilsson
authors.affiliation: From the Experimental Cardiovascular Research Unit, Clinical Research Centre, Clinical Sciences Malmö, Lund University, Malmö, Sweden (A.E., P.D., G.A., H.G., M.N., A.F.P., J.N., I.G.); Vascular Centre Malmö-Lund, Skåne, University Hospital, Malmö, Sweden (G.A.); Department of Cardiology, Skåne University Hospital, Lund/Malmö, Sweden (A.E., H.G., A.F.P., I.G.); Department of Molecular and Clinical Medicine/Wallenberg Laboratory, Sahlgrenska University Hospital University, Gothenburg, Sweden (M.S., J.B.); Cardiovascular Medicine Unit, Department of Medicine Solna, Karolinska Institutet, Solna, Sweden (R.M.F.); and Department of Clinical Sciences Malmö, Clinical Research Center, Lund University, Malmö, Sweden (I.G.M., O.M., M.O.-M.).
authors.forename: Isabel
authors.initials: I
authors.lastname: Gonçalves
mesh_terms: D000368:Aged; D017209:Apoptosis; D002339:Carotid Arteries / Q000378:metabolism / Q000473:pathology; D002340:Carotid Artery Diseases / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D003331:Coronary Vessels / Q000378:metabolism / Q000473:pathology; D016207:Cytokines / Q000378:metabolism; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D058226:Plaque, Atherosclerotic; D012422:Rupture, Spontaneous; D013107:Sphingolipids / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D016207:Cytokines; D013107:Sphingolipids; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: atherosclerosis; carotid stenosis; inflammation; sphingolipids; stroke