title: Netrin-1 promotes glioblastoma cell invasiveness and angiogenesis by multiple pathways including activation of RhoA, cathepsin B, and cAMP-response element-binding protein.
abstract: Glioblastomas are very difficult tumors to treat because they are highly invasive and disseminate within the normal brain, resulting in newly growing tumors. We have identified netrin-1 as a molecule that promotes glioblastoma invasiveness. As evidence, netrin-1 stimulates glioblastoma cell invasion directly through Matrigel-coated transwells, promotes tumor cell sprouting and enhances metastasis to lymph nodes in vivo. Furthermore, netrin-1 regulates angiogenesis as shown in specific angiogenesis assays such as enhanced capillary endothelial cells (EC) sprouting and by increased EC infiltration into Matrigel plugs in vivo, as does VEGF-A. This netrin-1 signaling pathway in glioblastoma cells includes activation of RhoA and cyclic AMP response element-binding protein (CREB). A novel finding is that netrin-1-induced glioblastoma invasiveness and angiogenesis are mediated by activated cathepsin B (CatB), a cysteine protease that translocates to the cell surface as an active enzyme and co-localizes with cell surface annexin A2 (ANXA2). The specific CatB inhibitor CA-074Me inhibits netrin-1-induced cell invasion, sprouting, and Matrigel plug angiogenesis. Silencing of CREB suppresses netrin-1-induced glioblastoma cell invasion, sprouting, and CatB expression. It is concluded that netrin-1 plays an important dual role in glioblastoma progression by promoting both glioblastoma cell invasiveness and angiogenesis in a RhoA-, CREB-, and CatB-dependent manner. Targeting netrin-1 pathways may be a promising strategy for brain cancer therapy.
authors.affiliation: Vascular Biology Program, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Akio
authors.initials: A
authors.lastname: Shimizu
authors.forename: Hironao
authors.initials: H
authors.lastname: Nakayama
authors.forename: Priscilla
authors.initials: P
authors.lastname: Wang
authors.forename: Courtney
authors.initials: C
authors.lastname: König
authors.forename: Tomoshige
authors.initials: T
authors.lastname: Akino
authors.forename: Johanna
authors.initials: J
authors.lastname: Sandlund
authors.forename: Silvia
authors.initials: S
authors.lastname: Coma
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Italiano
authors.forename: Akiko
authors.initials: A
authors.lastname: Mammoto
authors.forename: Diane R
authors.initials: DR
authors.lastname: Bielenberg
authors.forename: Michael
authors.initials: M
authors.lastname: Klagsbrun
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D017306:Annexin A2 / Q000378:metabolism; D002401:Cathepsin B / Q000378:metabolism*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D003094:Collagen / Q000737:chemistry; D000242:Cyclic AMP / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004338:Drug Combinations; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D005909:Glioblastoma / Q000378:metabolism*; D057809:HEK293 Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D008954:Models, Biological; D009361:Neoplasm Invasiveness; D009389:Neovascularization, Pathologic; D009414:Nerve Growth Factors / Q000378:metabolism*; D000075388:Netrin-1; D011485:Protein Binding; D011509:Proteoglycans / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D000199:Actins; D017306:Annexin A2; D017362:Cyclic AMP Response Element-Binding Protein; D004338:Drug Combinations; D007797:Laminin; C000617351:NTN1 protein, human; D009414:Nerve Growth Factors; C000617353:Ntn1 protein, mouse; D011509:Proteoglycans; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins; C059485:matrigel; C066737:RHOA protein, human; D000075388:Netrin-1; D003094:Collagen; D000242:Cyclic AMP; D002401:Cathepsin B; D020742:rhoA GTP-Binding Protein