title: Genetic deletion of platelet glycoprotein Ib alpha but not its extracellular domain protects from atherosclerosis.
abstract: The pathogenesis of atherosclerosis involves the interplay of haematopoietic, stromal and endothelial cells. Platelet interactions with endothelium and leukocytes are pivotal for atherosclerosis promotion. Glycoprotein (GP) Ibα is the ligand-binding subunit of the platelet GPIb-IX-V receptor complex; its deficiency causes the Bernard-Soulier syndrome (BSS), characterised by absent platelet GPIb-IX-V, macrothrombocytopenia and bleeding. We designed this study to determine the role of platelet GPIbα in the pathogenesis of atherosclerosis using two unique knockout models. Ldlr-/- mice were reconstituted with wild-type (wt), GPIbα-/- (lacks GPIbα) or chimeric IL-4R/GPIbα-Tg (lacks GPIbα extracellular domain) bone marrow and assayed for atherosclerosis development after feeding with pro-atherogenic "western diet". Here, we report that Ldlr-/-mice reconstituted with GPIbα-/- bone marrow developed less atherosclerosis compared to wt controls; accompanied by augmented accumulation of pro-inflammatory CD11b+ and CD11c+ myeloid cells, reduced oxLDL uptake and decreased TNF and IL 12p35 gene expression in the aortas. Flow cytometry and live cell imaging in whole blood-perfused microfluidic chambers revealed reduced platelet-monocyte aggregates in GPIbα-/- mice, which resulted in decreased monocyte activation. Interestingly, Ldlr-/-mice reconstituted with IL-4R/GPIbα-Tg bone marrow, producing less abnormal platelets, showed atherosclerotic lesions similar to wt mice. Platelet interaction with blood monocytes and accumulation of myeloid cells in the aortas were also essentially unaltered. Moreover, only complete GPIbα ablation altered platelet microparticles and CCL5 chemokine production. Thus, atherosclerosis reduction in mice lacking GPIbα may not result from the defective GPIbα-ligand binding, but more likely is a consequence of functional defects of GPIbα-/- platelets and reduced blood platelet counts.
authors.forename: E K
authors.initials: EK
authors.lastname: Koltsova
authors.forename: P
authors.initials: P
authors.lastname: Sundd
authors.forename: A
authors.initials: A
authors.lastname: Zarpellon
authors.forename: H
authors.initials: H
authors.lastname: Ouyang
authors.forename: Z
authors.initials: Z
authors.lastname: Mikulski
authors.forename: A
authors.initials: A
authors.lastname: Zampolli
authors.forename: Z M
authors.initials: ZM
authors.lastname: Ruggeri
authors.affiliation: Klaus Ley, MD, Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA, 92037, USA, Fax: +1 858 752 6985, E-mail: Klaus@liai.org.
authors.forename: K
authors.initials: K
authors.lastname: Ley
mesh_terms: D000818:Animals; D001018:Aortic Diseases / Q000097:blood / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D050197:Atherosclerosis / Q000097:blood / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D001606:Bernard-Soulier Syndrome / Q000235:genetics / Q000378:metabolism*; D001792:Blood Platelets / Q000378:metabolism; D016026:Bone Marrow Transplantation; D039481:CD11b Antigen / Q000378:metabolism; D039521:CD11c Antigen / Q000378:metabolism; D018946:Chemokine CCL5 / Q000378:metabolism; D066273:Diet, Western; D004195:Disease Models, Animal; D005260:Female; D018836:Inflammation Mediators / Q000378:metabolism; D053765:Interleukin-12 Subunit p35 / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000378:metabolism; D008297:Male; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009000:Monocytes / Q000378:metabolism; D010973:Platelet Adhesiveness; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D019948:Receptors, Interleukin-4 / Q000378:metabolism; D011973:Receptors, LDL / Q000172:deficiency / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D039481:CD11b Antigen; D039521:CD11c Antigen; C503460:Ccl5 protein, mouse; D018946:Chemokine CCL5; C508859:Il12a protein, mouse; D018836:Inflammation Mediators; D053765:Interleukin-12 Subunit p35; D008077:Lipoproteins, LDL; D019038:Platelet Glycoprotein GPIb-IX Complex; D019948:Receptors, Interleukin-4; D011973:Receptors, LDL; D014409:Tumor Necrosis Factor-alpha; C094990:adhesion receptor; C099252:oxidized low density lipoprotein
keywords: Aorta; atherosclerosis; inflammation; myeloid cells; platelets
references.citation: J Exp Med. 1999 Sep 20;190(6):803-14
references.pmid: 10499919
references.citation: Proc Natl Acad Sci U S A. 2011 May 24;108(21):8628-33
references.pmid: 21555542
references.citation: J Thromb Haemost. 2009 Nov;7(11):1935-8
references.pmid: 19740100
references.citation: Nat Med. 2003 Jan;9(1):61-7
references.pmid: 12483207
references.citation: J Cell Biol. 1989 Jul;109(1):367-75
references.pmid: 2545728
references.citation: Circ Res. 2011 Nov 11;109(11):1269-79
references.pmid: 21998326
references.citation: Curr Opin Hematol. 2007 May;14(3):262-9
references.pmid: 17414217
references.citation: Circulation. 2001 Apr 3;103(13):1772-7
references.pmid: 11282909
references.citation: Thromb Haemost. 2002 Aug;88(2):307-14
references.pmid: 12195705
references.citation: Microcirculation. 2011 Jul;18(5):361-72
references.pmid: 21418380
references.citation: Nat Med. 2009 Jan;15(1):97-103
references.pmid: 19122657
references.citation: Eur J Pharmacol. 2012 May 5;682(1-3):153-60
references.pmid: 22374261
references.citation: Circ Res. 2012 Apr 13;110(8):1036-8
references.pmid: 22499894
references.citation: Circulation. 2005 Aug 23;112(8):1180-8
references.pmid: 16103235
references.citation: Blood. 2003 Apr 1;101(7):2661-6
references.pmid: 12480714
references.citation: Blood. 2001 Sep 1;98(5):1424-8
references.pmid: 11520791
references.citation: Nat Med. 2002 Mar;8(3):247-52
references.pmid: 11875495
references.citation: Circulation. 1998 Sep 22;98(12):1164-71
references.pmid: 9743506
references.citation: J Biol Chem. 1986 Jan 5;261(1):381-5
references.pmid: 2934387
references.citation: Arterioscler Thromb Vasc Biol. 2011 Jul;31(7):1506-16
references.pmid: 21677293
references.citation: Arterioscler Thromb Vasc Biol. 2004 Jun;24(6):1124-9
references.pmid: 15072990
references.citation: Nature. 1998 Aug 13;394(6694):690-4
references.pmid: 9716134
references.citation: J Exp Med. 2000 Apr 17;191(8):1413-22
references.pmid: 10770806
references.citation: J Clin Invest. 2012 Sep;122(9):3114-26
references.pmid: 22886300
references.citation: Nature. 2012 Aug 16;488(7411):399-403
references.pmid: 22763437
references.citation: Arterioscler Thromb Vasc Biol. 2009 Oct;29(10):1419-23
references.pmid: 19696407
references.citation: Blood. 2013 May 30;121(22):4555-66
references.pmid: 23550035
references.citation: Immunity. 2003 Jul;19(1):71-82
references.pmid: 12871640
references.citation: J Clin Invest. 1983 Jul;72(1):1-12
references.pmid: 6223940
references.citation: Circulation. 2005 Nov 8;112(19):2993-3000
references.pmid: 16260637
references.citation: Circulation. 2002 May 7;105(18):2130-2
references.pmid: 11994242
references.citation: J Exp Med. 2002 Oct 7;196(7):887-96
references.pmid: 12370251
references.citation: Circ Res. 2011 Aug 19;109(5):593-606
references.pmid: 21852557
references.citation: Blood. 2002 Sep 15;100(6):2102-7
references.pmid: 12200373
references.citation: Am J Respir Crit Care Med. 2002 Jul 1;166(1):53-60
references.pmid: 12091171
references.citation: J Lipid Res. 2008 Oct;49(10):2113-23
references.pmid: 18599739
references.citation: Circulation. 2002 May 7;105(18):2166-71
references.pmid: 11994250
references.citation: J Exp Med. 1998 Feb 2;187(3):329-39
references.pmid: 9449713
references.citation: Semin Thromb Hemost. 2013 Sep;39(6):656-62
references.pmid: 23929303
references.citation: J Clin Invest. 2005 Dec;115(12):3378-84
references.pmid: 16322783
references.citation: Genesis. 2004 Dec;40(4):241-6
references.pmid: 15593332
references.citation: Nature. 1998 Feb 5;391(6667):591-4
references.pmid: 9468137
references.citation: Nat Methods. 2010 Oct;7(10):821-4
references.pmid: 20871617
references.citation: Heart. 2003 Oct;89(10):1273-8
references.pmid: 12975446
references.citation: Circulation. 2008 Jun 24;117(25):3227-37
references.pmid: 18519846
references.citation: Eur J Pharmacol. 2008 Sep 11;592(1-3):128-32
references.pmid: 18644365
references.citation: Circulation. 2003 May 6;107(17):2244-9
references.pmid: 12707243
references.citation: Int J Biochem Cell Biol. 2003 Aug;35(8):1170-4
references.pmid: 12757754
references.citation: J Exp Med. 2000 Jul 17;192(2):193-204
references.pmid: 10899906
references.citation: Nat Rev Cardiol. 2010 Feb;7(2):77-86
references.pmid: 20065951
references.citation: Nat Methods. 2007 Mar;4(3):219-22
references.pmid: 17322889
references.citation: Circ Res. 2012 Oct 26;111(10):1274-85
references.pmid: 22927332
references.citation: N Engl J Med. 2011 May 5;364(18):1746-60
references.pmid: 21542745
references.citation: Blood. 2011 Mar 3;117(9):2718-27
references.pmid: 21156842
references.citation: Cardiovasc Res. 2004 Feb 15;61(3):498-511
references.pmid: 14962480
references.citation: Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7450-4
references.pmid: 7543682
references.citation: Arterioscler Thromb Vasc Biol. 2007 Jan;27(1):241-7
references.pmid: 17095718
references.citation: Thromb Haemost. 2007 Mar;97(3):425-34
references.pmid: 17334510
references.citation: Blood. 1999 Dec 15;94(12):4112-21
references.pmid: 10590056
references.citation: J Immunol. 2010 Dec 1;185(11):7057-66
references.pmid: 21037096
references.citation: Nat Med. 2005 Nov;11(11):1167-9
references.pmid: 16258538
references.citation: Thromb Haemost. 2010 Nov;104(5):894-902
references.pmid: 20941453
references.citation: Proc Natl Acad Sci U S A. 2000 Mar 14;97(6):2803-8
references.pmid: 10706630
references.citation: J Thromb Haemost. 2010 Nov;8(11):2358-68
references.pmid: 20695980
references.citation: Atherosclerosis. 2000 Jan;148(1):75-85
references.pmid: 10580173
references.citation: Blood. 2002 Jun 15;99(12):4486-93
references.pmid: 12036879