title: Domains I and IV of annexin A2 affect the formation and integrity of in vitro capillary-like networks.
abstract: Annexin A2 (AnxA2) is a widely expressed multifunctional protein found in different cellular compartments. In spite of lacking a hydrophobic signal peptide, AnxA2 is found at the cell surface of endothelial cells, indicative of a role in angiogenesis. Increased extracellular levels of AnxA2 in tumours correlate with neoangiogenesis, metastasis and poor prognosis. We hypothesised that extracellular AnxA2 may contribute to angiogenesis by affecting endothelial cell-cell interactions and motility. To address this question, we studied the effect of heterotetrameric and monomeric forms of AnxA2, as well as its two soluble domains on the formation and maintenance of capillary-like structures by using an in vitro co-culture system consisting of endothelial and smooth muscle cells. In particular, addition of purified domains I and IV of AnxA2 potently inhibited the vascular endothelial growth factor (VEGF)-dependent formation of the capillary-like networks in a dose-dependent manner. In addition, these AnxA2 domains disrupted endothelial cell-cell contacts in preformed capillary-like networks, resulting in the internalisation of vascular endothelial (VE)-cadherin and the formation of VE-cadherin-containing filopodia-like structures between the endothelial cells, suggesting increased cell motility. Addition of monoclonal AnxA2 antibodies, in particular against Tyr23 phosphorylated AnxA2, also strongly inhibited network formation in the co-culture system. These results suggest that extracellular AnxA2, most likely in its Tyr phosphorylated form, plays a pivotal role in angiogenesis. The exogenously added AnxA2 domains most likely mediate their effects by competing with endogenous AnxA2 for extracellular factors necessary for the initiation and maintenance of angiogenesis, such as those involved in the formation/integrity of cell-cell contacts.
authors.affiliation: Department of Biomedicine, University of Bergen, Bergen, Norway.
authors.forename: Aase M
authors.initials: AM
authors.lastname: Raddum
authors.forename: Lasse
authors.initials: L
authors.lastname: Evensen
authors.forename: Hanne
authors.initials: H
authors.lastname: Hollås
authors.forename: Ann Kari
authors.initials: AK
authors.lastname: Grindheim
authors.forename: James B
authors.initials: JB
authors.lastname: Lorens
authors.forename: Anni
authors.initials: A
authors.lastname: Vedeler
mesh_terms: D000818:Animals; D017306:Annexin A2 / Q000378:metabolism* / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D018613:Microscopy, Confocal; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009418:S100 Proteins / Q000378:metabolism; D000099248:S100 Calcium Binding Protein A10
chemical_list: D017306:Annexin A2; D009418:S100 Proteins; D000099248:S100 Calcium Binding Protein A10
references.citation: Cancer Cell. 2009 Mar 3;15(3):167-70
references.pmid: 19249675
references.citation: J Mol Biol. 1998 Jun 26;279(5):1163-75
references.pmid: 9642092
references.citation: J Biol Chem. 1994 Aug 19;269(33):21191-7
references.pmid: 8063740
references.citation: J Biol Chem. 2004 Jun 18;279(25):26645-53
references.pmid: 15033986
references.citation: Biol Chem. 2012 Oct;393(10):1151-63
references.pmid: 23091278
references.citation: J Mol Biol. 2006 Oct 20;363(2):469-81
references.pmid: 16963080
references.citation: Mol Biol Cell. 1997 Mar;8(3):533-45
references.pmid: 9188103
references.citation: Nat Rev Mol Cell Biol. 2005 Jun;6(6):449-61
references.pmid: 15928709
references.citation: Biochem J. 2000 Jun 15;348 Pt 3:565-72
references.pmid: 10839987
references.citation: J Biol Chem. 1992 Jul 15;267(20):13993-7
references.pmid: 1385810
references.citation: Cancer Microenviron. 2011 Aug;4(2):199-208
references.pmid: 21909879
references.citation: Cell Cycle. 2006 Sep;5(18):2057-9
references.pmid: 16969103
references.citation: J Cell Sci. 2008 Jul 1;121(Pt 13):2115-22
references.pmid: 18565824
references.citation: J Cell Sci. 2004 Jun 1;117(Pt 13):2631-9
references.pmid: 15169834
references.citation: J Cell Sci. 2011 May 1;124(Pt 9):1453-64
references.pmid: 21486955
references.citation: J Cell Sci. 2002 Jan 1;115(Pt 1):91-8
references.pmid: 11801727
references.citation: Blood. 2010 Jan 21;115(3):549-58
references.pmid: 19965653
references.citation: Cancer Res. 2002 Jul 15;62(14):4015-22
references.pmid: 12124335
references.citation: Nat Med. 2000 Apr;6(4):389-95
references.pmid: 10742145
references.citation: J Biol Chem. 2011 Sep 2;286(35):30911-30925
references.pmid: 21737841
references.citation: EMBO J. 2006 May 3;25(9):1816-26
references.pmid: 16601677
references.citation: J Biol Chem. 2005 Dec 30;280(52):42831-40
references.pmid: 16230353
references.citation: Am J Physiol Cell Physiol. 2004 May;286(5):C987-97
references.pmid: 15075197
references.citation: Mol Vis. 2009 Jun 17;15:1231-42
references.pmid: 19536308
references.citation: Nat Rev Mol Cell Biol. 2004 Apr;5(4):317-23
references.pmid: 15071556
references.citation: Arterioscler Thromb Vasc Biol. 2008 Feb;28(2):223-32
references.pmid: 18162609
references.citation: Genome Biol. 2004;5(4):219
references.pmid: 15059252
references.citation: FASEB J. 2002 Aug;16(10):1177-84
references.pmid: 12153985
references.citation: Mol Cell Proteomics. 2012 Jul;11(7):M112.017988
references.pmid: 22493182
references.citation: Am J Physiol Gastrointest Liver Physiol. 2012 Apr;302(7):G712-22
references.pmid: 22241862
references.citation: PLoS One. 2009 Jun 04;4(6):e5798
references.pmid: 19495422
references.citation: Biochemistry. 2010 Mar 16;49(10):2216-26
references.pmid: 20121258
references.citation: Trends Cardiovasc Med. 1999 Apr-May;9(3-4):92-102
references.pmid: 10578524
references.citation: Science. 2005 Jan 7;307(5706):58-62
references.pmid: 15637262
references.citation: J Biol Chem. 2009 Jan 16;284(3):1604-11
references.pmid: 18990701
references.citation: N Engl J Med. 1999 Apr 1;340(13):994-1004
references.pmid: 10099141
references.citation: J Biol Chem. 2006 Oct 20;281(42):31689-95
references.pmid: 16882661
references.citation: Exp Cell Res. 1998 Feb 1;238(2):324-34
references.pmid: 9473340
references.citation: Vascul Pharmacol. 2011 Mar-Jun;54(3-6):59-67
references.pmid: 21440088
references.citation: Cancer Lett. 2007 Jul 8;252(1):19-35
references.pmid: 17188424
references.citation: Mol Cell Biol. 2008 Mar;28(5):1657-68
references.pmid: 18160703
references.citation: Biochim Biophys Acta. 1994 Apr 5;1197(1):63-93
references.pmid: 8155692
references.citation: Curr Opin Cell Biol. 2007 Oct;19(5):508-14
references.pmid: 17935963
references.citation: Exp Mol Pathol. 2012 Feb;92(1):175-84
references.pmid: 22044461
references.citation: Blood. 2011 Sep 8;118(10):2918-29
references.pmid: 21788340
references.citation: EMBO J. 2000 Mar 1;19(5):913-20
references.pmid: 10698933
references.citation: J Biol Chem. 2000 Oct 27;275(43):33189-92
references.pmid: 10961983
references.citation: Mol Cell Biol. 2008 Feb;28(3):1029-40
references.pmid: 18070928
references.citation: Cancer Res. 2012 Nov 1;72(21):5600-12
references.pmid: 22926560
references.citation: J Biol Chem. 2001 Feb 16;276(7):5310-5
references.pmid: 11067857
references.citation: J Biol Chem. 2003 Dec 12;278(50):50338-45
references.pmid: 14522961
references.citation: EMBO J. 1984 Jan;3(1):227-33
references.pmid: 6323166
references.citation: PLoS One. 2011 Apr 29;6(4):e19390
references.pmid: 21572519
references.citation: Pflugers Arch. 2008 Jan;455(4):575-82
references.pmid: 17638009
references.citation: Ann N Y Acad Sci. 1997 Apr 15;811:337-49
references.pmid: 9186609
references.citation: Acta Histochem. 2007;109(2):89-110
references.pmid: 17257660
references.citation: Circ Res. 2007 Mar 30;100(6):782-94
references.pmid: 17395884
references.citation: Dev Cell. 2011 Apr 19;20(4):526-39
references.pmid: 21396893
references.citation: J Cell Sci. 2008 Jul 1;121(Pt 13):2177-85
references.pmid: 18565825
references.citation: J Biol Chem. 2003 Feb 21;278(8):5702-9
references.pmid: 12468550
references.citation: Front Biosci. 2002 Feb 01;7:d341-8
references.pmid: 11815288
references.citation: J Cell Biochem. 1993 Nov;53(3):265-76
references.pmid: 8263043
references.citation: J Cell Biol. 1993 Mar;120(6):1357-69
references.pmid: 8449982
references.citation: J Cell Biol. 2004 Jan 5;164(1):133-44
references.pmid: 14699089
references.citation: Biochim Biophys Acta. 2006 Nov;1763(11):1325-34
references.pmid: 17045350
references.citation: J Cell Sci. 1997 Jan;110 ( Pt 2):221-8
references.pmid: 9044052
references.citation: Traffic. 2007 Aug;8(8):951-8
references.pmid: 17547702
references.citation: J Biol Chem. 1994 Aug 19;269(33):21198-203
references.pmid: 8063741
references.citation: Curr Pharm Des. 2010;16(35):3958-63
references.pmid: 21158729
references.citation: J Biol Chem. 2010 Dec 24;285(52):40624-34
references.pmid: 20947498
references.citation: FASEB J. 2001 Feb;15(2):447-57
references.pmid: 11156960
references.citation: Mol Biol Cell. 2012 Jan;23(2):310-23
references.pmid: 22090347
references.citation: Cytometry A. 2010 Jan;77(1):41-51
references.pmid: 19834964
references.citation: Int J Cell Biol. 2012;2012:852430
references.pmid: 22505935
references.citation: J Mol Biol. 2007 May 18;368(5):1367-78
references.pmid: 17395201
references.citation: PLoS One. 2007 Oct 31;2(10):e1118
references.pmid: 17971878
references.citation: Exp Mol Pathol. 2010 Apr;88(2):278-86
references.pmid: 20079732
references.citation: Mol Pharmacol. 2006 Nov;70(5):1469-80
references.pmid: 16887934
references.citation: Nat Med. 2003 Jun;9(6):653-60
references.pmid: 12778163
references.citation: Physiol Rev. 2002 Apr;82(2):331-71
references.pmid: 11917092