title: Netrin-4 induces lymphangiogenesis in vivo.
abstract: Netrin-4, a laminin-related secreted protein is an axon guidance cue recently shown essential outside of the nervous system, regulating mammary and lung morphogenesis as well as blood vascular development. Here, we show that Netrin-4, at physiologic doses, induces proliferation, migration, adhesion, tube formation and survival of human lymphatic endothelial cells in vitro comparable to well-characterized lymphangiogenic factors fibroblast growth factor-2 (FGF-2), hepatocyte growth factor (HGF), vascular endothelial growth factor-A (VEGF-A), and vascular endothelial growth factor-C (VEGF-C). Netrin-4 stimulates phosphorylation of intracellular signaling components Akt, Erk and S6, and their specific inhibition antagonizes Netrin-4-induced proliferation. Although Netrin receptors Unc5B and neogenin, are expressed by human lymphatic endothelial cells, suppression of either or both does not suppress Netrin-4-promoted in vitro effects. In vivo, Netrin-4 induces growth of lymphatic and blood vessels in the skin of transgenic mice and in breast tumors. Its overexpression in human and mouse mammary carcinoma cancer cells leads to enhanced metastasis. Finally, Netrin-4 stimulates in vitro and in vivo lymphatic permeability by activating small GTPases and Src family kinases/FAK, and down-regulating tight junction proteins. Together, these data provide evidence that Netrin-4 is a lymphangiogenic factor contributing to tumor dissemination and represents a potential target to inhibit metastasis formation.
authors.affiliation: Program in Molecular Medicine, Department of Medicine, University of Utah, Salt Lake City, UT, USA.
authors.forename: Frederic
authors.initials: F
authors.lastname: Larrieu-Lahargue
authors.forename: Alana L
authors.initials: AL
authors.lastname: Welm
authors.forename: Kirk R
authors.initials: KR
authors.lastname: Thomas
authors.forename: Dean Y
authors.initials: DY
authors.lastname: Li
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000098:blood supply / Q000235:genetics / Q000473:pathology; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D042583:Lymphangiogenesis; D042601:Lymphatic Vessels / Q000166:cytology* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism*; D000075386:Netrin Receptors; D000075385:Netrins; D011956:Receptors, Cell Surface / Q000378:metabolism; D012867:Skin / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; C498815:NTN4 protein, human; D009414:Nerve Growth Factors; D000075386:Netrin Receptors; D000075385:Netrins; C416214:Ntn4 protein, mouse; D011956:Receptors, Cell Surface; C532445:UNC5B protein, human; C091279:neogenin
references.citation: Proc Natl Acad Sci U S A. 2004 Nov 16;101(46):16210-5
references.pmid: 15520390
references.citation: EMBO J. 2005 Aug 17;24(16):2885-95
references.pmid: 16052207
references.citation: Biochim Biophys Acta. 2009 Apr;1795(2):137-51
references.pmid: 19162129
references.citation: Int J Cancer. 2009 Dec 15;125(12):2747-56
references.pmid: 19569051
references.citation: J Biol Chem. 2007 Aug 17;282(33):23750-8
references.pmid: 17588941
references.citation: Cancer Cell. 2008 Apr;13(4):331-42
references.pmid: 18394556
references.citation: Mol Cell Biol. 2008 Jun;28(12):4026-39
references.pmid: 18411305
references.citation: Science. 1997 May 30;276(5317):1423-5
references.pmid: 9162011
references.citation: J Exp Med. 2007 Oct 1;204(10):2349-62
references.pmid: 17846148
references.citation: Br J Cancer. 2009 Jul 21;101(2):327-34
references.pmid: 19568240
references.citation: J Cell Biol. 2004 Oct 25;167(2):223-9
references.pmid: 15504909
references.citation: Cardiovasc Res. 2001 Feb 16;49(3):507-21
references.pmid: 11166264
references.citation: J Cell Sci. 2008 Jul 1;121(Pt 13):2115-22
references.pmid: 18565824
references.citation: Cancer Cell. 2002 Apr;1(3):219-27
references.pmid: 12086857
references.citation: Gastroenterology. 2008 Oct;135(4):1248-57
references.pmid: 18692059
references.citation: J Cell Biol. 2000 Oct 16;151(2):221-34
references.pmid: 11038171
references.citation: Cancer Sci. 2007 May;98(5):726-33
references.pmid: 17425689
references.citation: J Clin Invest. 2004 Apr;113(7):1040-50
references.pmid: 15057311
references.citation: Exp Cell Res. 2009 Mar 10;315(5):784-94
references.pmid: 19094984
references.citation: Dev Cell. 2009 Apr;16(4):614-20
references.pmid: 19386270
references.citation: Cancer Biol Ther. 2007 May;6(5):743-8
references.pmid: 17387275
references.citation: Dev Cell. 2003 Mar;4(3):371-82
references.pmid: 12636918
references.citation: Proc Natl Acad Sci U S A. 2009 May 5;106(18):7595-600
references.pmid: 19383784
references.citation: Proc Natl Acad Sci U S A. 2002 Jun 25;99(13):8868-73
references.pmid: 12070340
references.citation: Nat Rev Mol Cell Biol. 2007 Apr;8(4):296-306
references.pmid: 17356579
references.citation: Proc Natl Acad Sci U S A. 2005 Oct 11;102(41):14729-34
references.pmid: 16203981
references.citation: Eur J Cancer. 2007 Jul;43(11):1748-54
references.pmid: 17570654
references.citation: Science. 2006 Aug 4;313(5787):640-4
references.pmid: 16809490
references.citation: Pathol Oncol Res. 2008 Sep;14(3):299-306
references.pmid: 18386163
references.citation: Int J Cancer. 2002 Apr 20;98(6):946-51
references.pmid: 11948478
references.citation: Cancer Res. 1992 Mar 15;52(6):1399-405
references.pmid: 1540948
references.citation: Eur J Cancer. 2004 Dec;40(18):2717-25
references.pmid: 15571953
references.citation: Proc Natl Acad Sci U S A. 2008 Aug 26;105(34):12491-6
references.pmid: 18719102
references.citation: FASEB J. 2002 Jul;16(9):922-34
references.pmid: 12087053
references.citation: Circ Res. 2004 Mar 19;94(5):664-70
references.pmid: 14739162