title: The ether lipid precursor hexadecylglycerol protects against Shiga toxins.
abstract: Shiga toxin-producing Escherichia coli bacteria cause hemorrhagic colitis and hemolytic uremic syndrome in humans. Currently, only supportive treatment is available for diagnosed patients. We show here that 24-h pretreatment with an ether lipid precursor, the alkylglycerol sn-1-O-hexadecylglycerol (HG), protects HEp-2 cells against Shiga toxin and Shiga toxin 2. Also the endothelial cell lines HMEC-1 and HBMEC are protected against Shiga toxins after HG pretreatment. In contrast, the corresponding acylglycerol, DL-α-palmitin, has no effect on Shiga toxicity. Although HG treatment provides a strong protection (~30 times higher IC₅₀) against Shiga toxin, only a moderate reduction in toxin binding was observed, suggesting that retrograde transport of the toxin from the plasma membrane to the cytosol is perturbed. Furthermore, endocytosis of Shiga toxin and retrograde sorting from endosomes to the Golgi apparatus remain intact, but transport from the Golgi to the endoplasmic reticulum is inhibited by HG treatment. As previously described, HG reduces the total level of all quantified glycosphingolipids to 50-70% of control, including the Shiga toxin receptor globotriaosylceramide (Gb3), in HEp-2 cells. In accordance with this, we find that interfering with Gb3 biosynthesis by siRNA-mediated knockdown of Gb3 synthase for 24 h causes a similar cytotoxic protection and only a moderate reduction in toxin binding (to 70% of control cells). Alkylglycerols, including HG, have been administered to humans for investigation of therapeutic roles in disorders where ether lipid biosynthesis is deficient, as well as in cancer therapy. Further studies may reveal if HG can also have a therapeutic potential in Shiga toxin-producing E. coli infections.
authors.affiliation: Centre for Cancer Biomedicine, Faculty of Medicine, University of Oslo, Oslo, Norway.
authors.forename: Jonas
authors.initials: J
authors.lastname: Bergan
authors.forename: Tore
authors.initials: T
authors.lastname: Skotland
authors.forename: Anne Berit Dyve
authors.initials: AB
authors.lastname: Lingelem
authors.forename: Roger
authors.initials: R
authors.lastname: Simm
authors.forename: Bjørn
authors.initials: B
authors.lastname: Spilsberg
authors.forename: Toril
authors.initials: T
authors.lastname: Lindbäck
authors.forename: Tuulia
authors.initials: T
authors.lastname: Sylvänne
authors.forename: Helena
authors.initials: H
authors.lastname: Simolin
authors.forename: Kim
authors.initials: K
authors.lastname: Ekroos
authors.forename: Kirsten
authors.initials: K
authors.lastname: Sandvig
mesh_terms: D001692:Biological Transport; D019921:Biotinylation; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D004986:Ether / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry*; D005995:Glyceryl Ethers / Q000737:chemistry*; D006028:Glycosphingolipids / Q000737:chemistry; D006056:Golgi Apparatus / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D008055:Lipids / Q000737:chemistry; D019308:Palmitic Acid / Q000737:chemistry; D022621:Shiga Toxin / Q000737:chemistry; D022603:Shiga Toxins / Q000737:chemistry*; D014281:Trihexosylceramides / Q000737:chemistry
chemical_list: D005995:Glyceryl Ethers; D006028:Glycosphingolipids; D008055:Lipids; D022603:Shiga Toxins; D014281:Trihexosylceramides; D004986:Ether; D019308:Palmitic Acid; C018549:globotriaosylceramide; D022621:Shiga Toxin; D005990:Glycerol; C004348:chimyl alcohol
references.citation: Clin J Am Soc Nephrol. 2010 Jul;5(7):1218-28
references.pmid: 20498239
references.citation: J Cell Biol. 1994 Jul;126(1):53-64
references.pmid: 8027186
references.citation: PLoS One. 2010 Feb 05;5(2):e9079
references.pmid: 20140095
references.citation: J Cell Sci. 2011 Jan 1;124(Pt 1):5-8
references.pmid: 21172818
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 20;106(42):17711-6
references.pmid: 19815513
references.citation: J Cell Sci. 2013 Jun 15;126(Pt 12):2641-55
references.pmid: 23591818
references.citation: Nature. 2007 Sep 6;449(7158):62-7
references.pmid: 17687330
references.citation: Nat Chem Biol. 2011 May;7(5):260-2
references.pmid: 21460830
references.citation: Toxicon. 2012 Nov;60(6):1085-107
references.pmid: 22960449
references.citation: Traffic. 2011 Nov;12(11):1634-47
references.pmid: 21810155
references.citation: Biochim Biophys Acta. 2006 Dec;1763(12):1511-26
references.pmid: 17027098
references.citation: Anal Chem. 2006 Sep 1;78(17):6202-14
references.pmid: 16944903
references.citation: J Cell Biol. 2001 Oct 29;155(3):369-80
references.pmid: 11673476
references.citation: Biochim Biophys Acta. 2012 Sep;1822(9):1442-52
references.pmid: 22627108
references.citation: J Lipid Res. 2003 Jan;44(1):182-92
references.pmid: 12518037
references.citation: J Cell Sci. 2007 Jun 15;120(Pt 12):2010-21
references.pmid: 17550970
references.citation: Prog Lipid Res. 2014 Apr;54:1-13
references.pmid: 24462587
references.citation: J Biol Chem. 2007 Jun 1;282(22):16317-28
references.pmid: 17403690
references.citation: Biochem Biophys Res Commun. 1973 Mar 5;51(1):119-24
references.pmid: 4349319
references.citation: Glycoconj J. 1996 Apr;13(2):159-66
references.pmid: 8737240
references.citation: Lipids. 1992 Jun;27(6):401-5
references.pmid: 1630273
references.citation: PLoS One. 2012;7(3):e31413
references.pmid: 22403614
references.citation: Proc Jpn Acad Ser B Phys Biol Sci. 2013;89(4):129-38
references.pmid: 23574804
references.citation: Anal Chem. 2002 Mar 1;74(5):941-9
references.pmid: 11924996
references.citation: Nature. 1992 Aug 6;358(6386):510-2
references.pmid: 1641040
references.citation: Biochemistry. 2002 Feb 12;41(6):2075-88
references.pmid: 11827555
references.citation: Toxicon. 2010 Dec 15;56(7):1181-5
references.pmid: 19951719
references.citation: Mol Biol Cell. 2001 Aug;12(8):2453-68
references.pmid: 11514628
references.citation: Arch Immunol Ther Exp (Warsz). 2011 Aug;59(4):239-47
references.pmid: 21644029
references.citation: Curr Top Microbiol Immunol. 2012;357:19-40
references.pmid: 21761287
references.citation: Biochimie. 2013 Jan;95(1):59-65
references.pmid: 22771767
references.citation: Methods Enzymol. 1982;83:357-62
references.pmid: 6178935
references.citation: Kidney Int Suppl. 2009 Feb;(112):S62-6
references.pmid: 19180140
references.citation: Vaccine. 2006 Feb 20;24(8):1142-8
references.pmid: 16198455
references.citation: Mar Drugs. 2010 Aug 05;8(8):2267-300
references.pmid: 20948908
references.citation: Traffic. 2006 Sep;7(9):1243-53
references.pmid: 16919154
references.citation: PLoS One. 2013 Sep 30;8(9):e75904
references.pmid: 24098742
references.citation: Pol Merkur Lekarski. 2005 Jun;18(108):686-92
references.pmid: 16124384
references.citation: J Lipid Res. 1988 Mar;29(3):325-34
references.pmid: 3379344
references.citation: FEBS J. 2006 Nov;273(22):5205-18
references.pmid: 17059464
references.citation: Curr Top Microbiol Immunol. 2012;357:67-103
references.pmid: 21915773
references.citation: Biochim Biophys Acta. 2012 Aug;1821(8):1104-13
references.pmid: 22281700
references.citation: Exp Hematol. 2000 Nov;28(11):1260-8
references.pmid: 11063874
references.citation: Microb Biotechnol. 2011 Jan;4(1):32-46
references.pmid: 21255370
references.citation: Kidney Int. 2009 Jun;75(11):1209-1216
references.pmid: 19212418
references.citation: J Cell Sci. 2004 Aug 1;117(Pt 17):3911-22
references.pmid: 15265987
references.citation: FASEB J. 2006 Dec;20(14):2597-9
references.pmid: 17065223
references.citation: Biochem Biophys Res Commun. 2009 Dec 4;390(1):109-14
references.pmid: 19782049
references.citation: J Biol Chem. 1999 Dec 3;274(49):35278-82
references.pmid: 10575015
references.citation: Traffic. 2009 Jul;10(7):868-82
references.pmid: 19453975
references.citation: FEBS Lett. 2010 May 3;584(9):1879-86
references.pmid: 19948172
references.citation: Cell. 2010 Apr 16;141(2):231-42
references.pmid: 20403321
references.citation: PLoS One. 2011;6(12):e28539
references.pmid: 22163031
references.citation: Science. 2012 Jan 20;335(6066):332-5
references.pmid: 22267811
references.citation: J Clin Microbiol. 1999 Mar;37(3):497-503
references.pmid: 9986802
references.citation: Methods. 2005 Jun;36(2):207-24
references.pmid: 15894491
references.citation: Mol Biol Cell. 2006 Mar;17(3):1375-87
references.pmid: 16381816
references.citation: Traffic. 2012 Mar;13(3):443-54
references.pmid: 22132807
references.citation: Prog Lipid Res. 2001 May;40(3):199-229
references.pmid: 11275267
references.citation: Dev Cell. 2012 Nov 13;23(5):886-95
references.pmid: 23153485
references.citation: Nat Protoc. 2007;2(9):2159-65
references.pmid: 17853872
references.citation: Biochim Biophys Acta. 2011 Nov;1811(11):925-34
references.pmid: 21767661
references.citation: Traffic. 2013 Jul;14(7):839-52
references.pmid: 23593995
references.citation: J Cell Physiol. 2008 Sep;216(3):750-63
references.pmid: 18446787
references.citation: Cell Mol Life Sci. 2014 Mar;71(6):1097-116
references.pmid: 23921715
references.citation: Biochim Biophys Acta. 2002 Nov 8;1585(1):39-43
references.pmid: 12457713
references.citation: J Biol Chem. 2010 Nov 12;285(46):36049-59
references.pmid: 20716521
references.citation: J Cell Physiol. 1998 Dec;177(4):646-60
references.pmid: 10092217
references.citation: Cell Microbiol. 2003 Feb;5(2):85-97
references.pmid: 12580945
references.citation: Histochem Cell Biol. 2013 Sep;140(3):317-26
references.pmid: 23765164
references.citation: Biochim Biophys Acta. 2008 Oct;1783(10):1857-65
references.pmid: 18571506
references.citation: Biochem Biophys Res Commun. 2007 Jun 29;358(2):566-70
references.pmid: 17498660
references.citation: Curr Top Microbiol Immunol. 2012;357:1-18
references.pmid: 21910078
references.citation: Future Microbiol. 2010 Mar;5(3):431-53
references.pmid: 20210553