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To identify the specific proteases responsible for MV formation, we induced angiogenesis in mouse tissues with an adenoviral vector expressing VEGF-A164 (Ad-VEGF-A164) or with VEGF-A\u2013secreting TA3\/St mammary tumors. We found that MV formation resulted from greatly increased activity of cathepsins (B&amp;gt;S&amp;gt;L) in venules transitioning into MV, as well as from a reciprocal decrease in the expression of several cysteine protease inhibitors (CPI), stefin A and cystatins B and C, by these same venules. Using a fluorescence probe that selectively binds cellular sites of cathepsin protease activity in vivo, we showed that increased cathepsin activity was localized exclusively to perivenular cells, not to venule endothelial cells. CPI strikingly inhibited angiogenesis in the Matrigel assay, and Ad-VEGF-A164\u2013induced angiogenesis was reduced by \u223c50% in cathepsin B\u2013null mice. Thus, VEGF-A, whether expressed by interstitial cells infected with an adenoviral vector or by tumor cells, upsets the normal cathepsin-CPI balance in nearby venules, leading to degradation of their basement membranes, an important first step in angiogenesis. [Cancer Res 2009;69(10):4537\u201344]<\/jats:p>","DOI":"10.1158\/0008-5472.can-08-4539","type":"journal-article","created":{"date-parts":[[2009,5,12]],"date-time":"2009-05-12T22:43:51Z","timestamp":1242168231000},"page":"4537-4544","update-policy":"https:\/\/doi.org\/10.1158\/crossmark_policy","source":"Crossref","is-referenced-by-count":105,"title":["VEGF-A Induces Angiogenesis by Perturbing the Cathepsin-Cysteine Protease Inhibitor Balance in Venules, Causing Basement Membrane Degradation and Mother Vessel Formation"],"prefix":"10.1158","volume":"69","author":[{"given":"Sung-Hee","family":"Chang","sequence":"first","affiliation":[{"name":"1Pathology and Departments of"}]},{"given":"Keizo","family":"Kanasaki","sequence":"additional","affiliation":[{"name":"2Medicine, and the Center for Vascular Biology Research, Beth Israel Deaconess Medical Center and Harvard Medical School, and"}]},{"given":"Vasilena","family":"Gocheva","sequence":"additional","affiliation":[{"name":"4Cancer Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York, New York; and"}]},{"given":"Galia","family":"Blum","sequence":"additional","affiliation":[{"name":"5Department of Pathology, Stanford University, Stanford, California"}]},{"given":"Jay","family":"Harper","sequence":"additional","affiliation":[{"name":"3Departments of Surgery, Children's Hospital and Harvard Medical School, Boston, Massachusetts;"}]},{"given":"Marsha A.","family":"Moses","sequence":"additional","affiliation":[{"name":"3Departments of Surgery, Children's Hospital and Harvard Medical School, Boston, Massachusetts;"}]},{"given":"Shou-Ching","family":"Shih","sequence":"additional","affiliation":[{"name":"1Pathology and Departments of"}]},{"given":"Janice A.","family":"Nagy","sequence":"additional","affiliation":[{"name":"1Pathology and Departments of"}]},{"given":"Johanna","family":"Joyce","sequence":"additional","affiliation":[{"name":"4Cancer Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York, New York; and"}]},{"given":"Matthew","family":"Bogyo","sequence":"additional","affiliation":[{"name":"5Department of Pathology, Stanford University, Stanford, California"}]},{"given":"Raghu","family":"Kalluri","sequence":"additional","affiliation":[{"name":"2Medicine, and the Center for Vascular Biology Research, Beth Israel Deaconess Medical Center and Harvard Medical School, and"}]},{"given":"Harold F.","family":"Dvorak","sequence":"additional","affiliation":[{"name":"1Pathology and Departments of"}]}],"member":"1086","published-online":{"date-parts":[[2009,5,15]]},"reference":[{"key":"2022061700291469800_B1","doi-asserted-by":"crossref","unstructured":"Folkman J. 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