{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,17]],"date-time":"2025-09-17T16:07:36Z","timestamp":1758125256468},"reference-count":26,"publisher":"Portland Press Ltd.","issue":"6","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2009,12,1]]},"abstract":"<jats:p>We have applied search algorithms to expression databases to identify genes whose expression is restricted to the endothelial cell. Such genes frequently play a critical role in endothelial biology and angiogenesis. Two such genes are the roundabout receptor Robo4 and the ECSCR (endothelial-cell-specific chemotaxis regulator). Endothelial cells express both Robo1 and Robo4, which we have knocked down using siRNA (small interfering RNA) and then studied the effect in a variety of in vitro assays. Both Robo4 and Robo1 knockdown inhibited in vitro tube formation on Matrigel\u2122. Transfection of Robo4 into endothelial cells increased the number of filopodial extensions from the cell, but failed to do so in Robo1-knockdown cells. Separate immunoprecipitation studies showed that Robo1 and Robo4 heterodimerize. We conclude from this and other work that a heteroduplex of Robo1 and Robo4 signals through WASP (Wiskott\u2013Aldrich syndrome protein) and other actin nucleation-promoting factors to increase the number of filopodia and cell migration. Knockdown of the transmembrane ECSCR protein in endothelial cells also reduced chemotaxis and impaired tube formation on Matrigel\u2122. Yeast two-hybrid analysis and immunoprecipitation studies showed that, in contrast with the roundabouts, ECSCR binds to the actin-modulatory filamin A. We conclude that all three of these genes are critical for effective endothelial cell migration and, in turn, angiogenesis.<\/jats:p>","DOI":"10.1042\/bst0371214","type":"journal-article","created":{"date-parts":[[2009,11,19]],"date-time":"2009-11-19T18:02:29Z","timestamp":1258653749000},"page":"1214-1217","update-policy":"http:\/\/dx.doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Functionally defining the endothelial transcriptome, from Robo4 to ECSCR"],"prefix":"10.1042","volume":"37","author":[{"given":"Ana\u00a0Raquel","family":"Verissimo","sequence":"first","affiliation":[{"name":"Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John\u00a0M.J.","family":"Herbert","sequence":"additional","affiliation":[{"name":"Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Victoria\u00a0L.","family":"Heath","sequence":"additional","affiliation":[{"name":"Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John\u00a0A.","family":"Legg","sequence":"additional","affiliation":[{"name":"Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Helen","family":"Sheldon","sequence":"additional","affiliation":[{"name":"Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maud","family":"Andre","sequence":"additional","affiliation":[{"name":"Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rajeeb\u00a0Kumar","family":"Swain","sequence":"additional","affiliation":[{"name":"Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roy","family":"Bicknell","sequence":"additional","affiliation":[{"name":"Institute for Biomedical Research, Birmingham University Medical School, Vincent Drive, Birmingham B15 2TT, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2009,11,19]]},"reference":[{"key":"2021111719312835200_B1","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/386671a0","article-title":"Mechanisms of angiogenesis","volume":"386","author":"Risau","year":"1997","journal-title":"Nature"},{"key":"2021111719312835200_B2","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/j.vph.2006.01.005","article-title":"Angiogenesis: a new target for future therapy","volume":"44","author":"Pandya","year":"2006","journal-title":"Vasc. Pharmacol."},{"key":"2021111719312835200_B3","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1136\/jcp.45.2.143","article-title":"Heterogeneity of vascular endothelial cells with relevance to diagnosis of vascular tumours","volume":"45","author":"Kuzu","year":"2002","journal-title":"J. Clin. Pathol."},{"key":"2021111719312835200_B4","doi-asserted-by":"crossref","first-page":"1796","DOI":"10.1101\/gr.150700","article-title":"In silico cloning of novel endothelial-specific genes","volume":"10","author":"Huminiecki","year":"2000","journal-title":"Genome Res."},{"key":"2021111719312835200_B5","doi-asserted-by":"crossref","first-page":"47079","DOI":"10.1074\/jbc.M308124200","article-title":"EndoPDI, a novel protein-disulfide isomerase-like protein that is preferentially expressed in endothelial cells acts as a stress survival factor","volume":"278","author":"Sullivan","year":"2003","journal-title":"J. Biol. Chem."},{"key":"2021111719312835200_B6","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1006\/geno.2002.6745","article-title":"Magic roundabout is a new member of the roundabout receptor family that endothelial specific and expressed at sites of active angiogenesis","volume":"79","author":"Huminiecki","year":"2002","journal-title":"Genomics"},{"key":"2021111719312835200_B7","doi-asserted-by":"crossref","first-page":"1712","DOI":"10.1161\/01.RES.0000269779.10644.dc","article-title":"A three-kilobase fragment of the human Robo4 promoter directs cell type-specific expression in endothelium","volume":"100","author":"Okada","year":"2007","journal-title":"Circ. Res."},{"key":"2021111719312835200_B8","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1016\/j.bbrc.2005.03.250","article-title":"Magic roundabout, a tumour endothelial marker: expression and signalling","volume":"332","author":"Seth","year":"2005","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2021111719312835200_B9","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/S1535-6108(03)00164-8","article-title":"Induction of tumor angiogenesis by Slit\u2013Robo signaling and inhibition of cancer growth by blocking Robo activity","volume":"4","author":"Wang","year":"2003","journal-title":"Cancer Cell"},{"key":"2021111719312835200_B10","first-page":"1437","article-title":"Robo1\/Robo4: differential expression of angiogenic markers in colorectal cancer","volume":"15","author":"Gr\u00f6ne","year":"2006","journal-title":"Oncol. Rep."},{"key":"2021111719312835200_B11","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1634\/stemcells.2008-0292","article-title":"Roundabout 4 is expressed in hematopoietic stem cells and potentially involved in the niche-mediated regulation of side population phenotype","volume":"27","author":"Shibata","year":"2009","journal-title":"Stem Cells"},{"key":"2021111719312835200_B12","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/S0012-1606(03)00258-6","article-title":"Robo4 is a vascular-specific receptor that inhibits endothelial migration","volume":"261","author":"Park","year":"2003","journal-title":"Dev. Biol."},{"key":"2021111719312835200_B13","doi-asserted-by":"crossref","first-page":"6373","DOI":"10.1073\/pnas.0408318102","article-title":"Roundabout4 is essential for angiogenesis in vivo","volume":"102","author":"Bedell","year":"2005","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021111719312835200_B14","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1038\/nm1742","article-title":"Robo4 stabilizes the vascular network by inhibiting pathologic angiogenesis and endothelial hyperpermeability","volume":"14","author":"Jones","year":"2008","journal-title":"Nat. Med."},{"key":"2021111719312835200_B15","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1038\/nbt0508-528","article-title":"Retinal anti-angiogenesis by a new route","volume":"26","author":"Bicknell","year":"2008","journal-title":"Nat. Biotechnol."},{"key":"2021111719312835200_B16","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1096\/fj.04-1991fje","article-title":"Soluble Robo4 receptor inhibits in vivo angiogenesis and endothelial cell migration","volume":"19","author":"Suchting","year":"2005","journal-title":"FASEB J."},{"key":"2021111719312835200_B17","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1186\/1471-2121-9-61","article-title":"Silencing of directional migration in roundabout4 knock down endothelial cells","volume":"9","author":"Kaur","year":"2008","journal-title":"BMC Cell Biol."},{"key":"2021111719312835200_B18","first-page":"1057","article-title":"Expression of Robo4 in the fibrovascular membranes from patients with proliferative diabetic retinopathy and its role in RF\/6A and RPE cells","volume":"15","author":"Huang","year":"2009","journal-title":"Mol. Vision"},{"key":"2021111719312835200_B19","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1096\/fj.07-098269","article-title":"Active involvement of Robo1 and Robo4 in filopodia formation and endothelial cell motility mediated via WASP and other actin nucleation-promoting factors","volume":"23","author":"Sheldon","year":"2009","journal-title":"FASEB J."},{"key":"2021111719312835200_B20","doi-asserted-by":"crossref","first-page":"1645","DOI":"10.1016\/S0960-9822(01)00506-1","article-title":"Cdc42 induces filopodia by promoting the formation of an IRSp53:Mena complex","volume":"11","author":"Krugmann","year":"2001","journal-title":"Curr. Biol."},{"key":"2021111719312835200_B21","doi-asserted-by":"crossref","first-page":"11347","DOI":"10.1074\/jbc.M508853200","article-title":"Robo4 signaling in endothelial cells implies attraction guidance mechanisms","volume":"281","author":"Kaur","year":"2006","journal-title":"J. Biol. Chem."},{"key":"2021111719312835200_B22","doi-asserted-by":"crossref","first-page":"6932","DOI":"10.1093\/emboj\/17.23.6932","article-title":"WAVE, a novel WASP-family protein involved in actin reorganization induced by Rac","volume":"17","author":"Miki","year":"1998","journal-title":"EMBO J."},{"key":"2021111719312835200_B23","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1186\/1471-2164-9-153","article-title":"A novel method of differential gene expression analysis using multiple cDNA libraries applied to the identification of tumour endothelial genes","volume":"9","author":"Herbert","year":"2008","journal-title":"BMC Genomics"},{"key":"2021111719312835200_B24","doi-asserted-by":"crossref","first-page":"1640","DOI":"10.1161\/ATVBAHA.108.162511","article-title":"ECSM2, an endothelial specific filamin A binding protein that mediates chemotaxis","volume":"28","author":"Armstrong","year":"2008","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"2021111719312835200_B25","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1111\/j.1365-2443.2008.01267.x","article-title":"Endothelial cell-specific molecule 2 (ECSM2) modulates actin remodelling and epidermal growth factor receptor signalling","volume":"14","author":"Ma","year":"2009","journal-title":"Genes Cells"},{"key":"2021111719312835200_B26","doi-asserted-by":"crossref","first-page":"8227","DOI":"10.1073\/pnas.0806780106","article-title":"Identification of ARIA regulating endothelial apoptosis and angiogenesis by modulating proteasomal degradation of cIAP-1 and cIAP-2","volume":"106","author":"Ikeda","year":"2009","journal-title":"Proc. Nat. Acad. Sci. U.S.A."}],"container-title":["Biochemical Society Transactions"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemsoctrans\/article-pdf\/37\/6\/1214\/546854\/bst0371214.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemsoctrans\/article-pdf\/37\/6\/1214\/546854\/bst0371214.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,17]],"date-time":"2021-11-17T21:44:01Z","timestamp":1637185441000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemsoctrans\/article\/37\/6\/1214\/64938\/Functionally-defining-the-endothelial"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,11,19]]},"references-count":26,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2009,11,19]]},"published-print":{"date-parts":[[2009,12,1]]}},"URL":"https:\/\/doi.org\/10.1042\/bst0371214","relation":{},"ISSN":["0300-5127","1470-8752"],"issn-type":[{"value":"0300-5127","type":"print"},{"value":"1470-8752","type":"electronic"}],"subject":[],"published":{"date-parts":[[2009,11,19]]}}}