The interaction of uPAR with VEGFR2 promotes VEGF-induced angiogenesis.

[en] In endothelial cells, binding of vascular endothelial growth factor (VEGF) to the receptor VEGFR2 activates multiple signaling pathways that trigger processes such as proliferation, survival, and migration that are necessary for angiogenesis. VEGF-bound VEGFR2 becomes internalized, which is a key step in the proangiogenic signal. We showed that the urokinase plasminogen activator receptor (uPAR) interacted with VEGFR2 and described the mechanism by which this interaction mediated VEGF signaling and promoted angiogenesis. Knockdown of uPAR in human umbilical vein endothelial cells (HUVECs) impaired VEGFR2 signaling, and uPAR deficiency in mice prevented VEGF-induced angiogenesis. Upon exposure of HUVECs to VEGF, uPAR recruited the low-density lipoprotein receptor-related protein 1 (LRP-1) to VEGFR2, which induced VEGFR2 internalization. Thus, the uPAR-VEGFR2 interaction is crucial for VEGF signaling in endothelial cells.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Herkenne, Stéphanie ; Université de Liège - ULiège > Cancer-Molecular Angiogenesis Laboratory

Paques, Cécile ; Université de Liège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire

Nivelles, Olivier

Lion, Michelle

BAJOU, Khalid ; Centre Hospitalier Universitaire de Liège - CHU > Centre d'oncologie

Pollenus, Thomas ; Université de Liège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire

Fontaine, Marie ; Université de Liège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire

Carmeliet, Peter

Martial, Joseph ; Université de Liège > Département des sciences de la vie > Département des sciences de la vie

Nguyen, Ngoc-Quynh-Nhu

Struman, Ingrid ; Université de Liège - ULiège

Language :

English

Title :

The interaction of uPAR with VEGFR2 promotes VEGF-induced angiogenesis.

Publication date :

2015

Journal title :

Science Signaling

ISSN :

1945-0877

eISSN :

1937-9145

Publisher :

American Association for the Advancement of Science, Washington, United States - District of Columbia

Volume :

Issue :

403

Pages :

ra117

Peer reviewed :

Peer Reviewed verified by ORBi

Commentary :

Available on ORBi :

since 11 December 2015

Statistics

Number of views

546 (58 by ULiège)

Number of downloads

444 (27 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

Bibliography

P. Carmeliet, Angiogenesis in life, disease and medicine. Nature 438, 932-936 (2005).
I. Zachary, Signal transduction in angiogenesis. EXS, 267-300 (2005).
J. Waltenberger, L. Claesson-Welsh, A. Siegbahn, M. Shibuya, C. H. Heldin, Different signal transduction properties of KDR and Flt1, two receptors for vascular endothelial growth factor. J. Biol. Chem. 269, 26988-26995 (1994).
S. Koch, Neuropilin signalling in angiogenesis. Biochem. Soc. Trans. 40, 20-25 (2012).
S. Koch, L. Claesson-Welsh, Signal transduction by vascular endothelial growth factor receptors. Cold Spring Harb. Perspect. Med. 2, a006502 (2012).
S. Koch, S. Tugues, X. Li, L. Gualandi, L. Claesson-Welsh, Signal transduction by vascular endothelial growth factor receptors. Biochem. J. 437, 169-183 (2011).
A. J. Pötgens, N. H. Lubsen,M. C. van Altena, R. Vermeulen, A. Bakker, J. G.G. Schoenmakers, D. J. Ruiter, R.M.W. deWaal, Covalent dimerization of vascular permeability factor/vascular endothelial growth factor is essential for its biological activity. Evidence from Cys to Ser mutations. J. Biol. Chem. 269, 32879-32885 (1994).
M. G. Blanes, M. Oubaha, Y. Rautureau, J.-P. Gratton, Phosphorylation of tyrosine 801 of vascular endothelial growth factor receptor-2 is necessary for Akt-dependent endothelial nitric-oxide synthase activation and nitric oxide release from endothelial cells. J. Biol. Chem. 282, 10660-10669 (2007).
T. Matsumoto, S. Bohman, J. Dixelius, T. Berge, A. Dimberg, P. Magnusson, L. Wang, C. Wikner, J. H. Qi, C. Wernstedt, J. Wu, S. Bruheim, H. Mugishima, D. Mukhopadhyay, A. Spurkland, L. Claesson-Welsh, VEGF receptor-2 Y951 signaling and a role for the adapter molecule TSAd in tumor angiogenesis. EMBO J. 24, 2342-2353 (2005).
K. Holmqvist, M. J. Cross, C. Rolny, R. Hägerkvist, N. Rahimi, T. Matsumoto, L. Claesson-Welsh, M. Welsh, The adaptor protein shb binds to tyrosine 1175 in vascular endothelial growth factor (VEGF) receptor-2 and regulates VEGF-dependent cellular migration. J. Biol. Chem. 279, 22267-22275 (2004).
T. Takahashi, H. Ueno, M. Shibuya, VEGF activates protein kinase C-dependent, but Ras-independent Raf-MEK-MAP kinase pathway for DNA synthesis in primary endothelial cells. Oncogene 18, 2221-2230 (1999).
T. Takahashi, S. Yamaguchi, K. Chida, M. Shibuya, A single autophosphorylation site on KDR/Flk-1 is essential for VEGF-A-dependent activation of PLC-g and DNA synthesis in vascular endothelial cells. EMBO J. 20, 2768-2778 (2001).
Y. Sakurai, K. Ohgimoto, Y. Kataoka, N. Yoshida, M. Shibuya, Essential role of Flk-1 (VEGF receptor 2) tyrosine residue 1173 in vasculogenesis in mice. Proc. Natl. Acad. Sci. U.S.A. 102, 1076-1081 (2005).
H.-P. Gerber, V. Dixit, N. Ferrara, Vascular endothelial growth factor induces expression of the antiapoptotic proteins Bcl-2 and A1 in vascular endothelial cells. J. Biol. Chem. 273, 13313-13316 (1998).
D. Fulton, J.-P. Gratton, T. J. McCabe, J. Fontana, Y. Fujio, K. Walsh, T. F. Franke, A. Papapetropoulos, W. C. Sessa, Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature 399, 597-601 (1999).
A. Eichmann, M. Simons, VEGF signaling inside vascular endothelial cells and beyond. Curr. Opin. Cell Biol. 24, 188-193 (2012).
M. G. Lampugnani, F. Orsenigo, M. C. Gagliani, C. Tacchetti, E. Dejana, Vascular endothelial cadherin controls VEGFR-2 internalization and signaling from intracellular compartments. J. Cell Biol. 174, 593-604 (2006).
R. D. Balsara, R. Merryman, F. Virjee, C. Northway, F. J. Castellino, V. A. Ploplis, A deficiency of uPAR alters endothelial angiogenic function and cell morphology. Vasc. Cell 3, 10 (2011).
M. Del Rosso, uPAR in angiogenesis regulation. Blood 117, 3941-3943 (2011).
M. Ploug, E. Rønne, N. Behrendt, A. L. Jensen, F. Blasi, K. Danø, Cellular receptor for urokinase plasminogen activator. Carboxyl-terminal processing and membrane anchoring by glycosyl-phosphatidylinositol. J. Biol. Chem. 266, 1926-1933 (1991).
A. Abdollahi, J. Folkman, Evading tumor evasion: Current concepts and perspectives of anti-angiogenic cancer therapy. Drug Resist. Updat. 13, 16-28 (2010).
M. P. Stoppelli, C. Tacchetti, M. V. Cubellis, A. Corti, V. J. Hearing, G. Cassani, E. Appella, F. Blasi, Autocrine saturation of pro-urokinase receptors on human A431 cells. Cell 45, 675-684 (1986).
G. W. Prager, J. M. Breuss, S. Steurer, J. Mihaly, B. R. Binder, Vascular endothelial growth factor (VEGF) induces rapid prourokinase (pro-uPA) activation on the surface of endothelial cells. Blood 103, 955-962 (2004).
G. W. Prager, J. M. Breuss, S. Steurer, D. Olcaydu, J. Mihaly, P. M. Brunner, H. Stockinger, B. R. Binder, Vascular endothelial growth factor receptor-2-induced initial endothelial cell migration depends on the presence of the urokinase receptor. Circ. Res. 94, 1562-1570 (2004).
R. A. Alexander, G. W. Prager, J. Mihaly-Bison, P. Uhrin, S. Sunzenauer, B. R. Binder, G. J. Schütz, M. Freissmuth, J. M. Breuss, VEGF-induced endothelial cell migration requires urokinase receptor (uPAR)-dependent integrin redistribution. Cardiovasc. Res. 94, 125-135 (2012).
C. D. Madsen, N. Sidenius, The interaction between urokinase receptor and vitronectin in cell adhesion and signalling. Eur. J. Cell Biol. 87, 617-629 (2008).
J. Kiyan, G. Smith, H. Haller, I. Dumler, Urokinase-receptor-mediated phenotypic changes in vascular smooth muscle cells require the involvement of membrane rafts. Biochem. J. 423, 343-351 (2009).
M. Resnati, I. Pallavicini, J. M. Wang, J. Oppenheim, C. N. Serhan, M. Romano, F. Blasi, The fibrinolytic receptor for urokinase activates the G protein-coupled chemotactic receptor FPRL1/LXA4R. Proc. Natl. Acad. Sci. U.S.A. 99, 1359-1364 (2002).
G. A. LaRusch, F. Mahdi, Z. Shariat-Madar, G. Adams, R. G. Sitrin, W. M. Zhang, K. R. McCrae, A. H. Schmaier, Factor XII stimulates ERK1/2 and Akt through uPAR, integrins, and the EGFR to initiate angiogenesis. Blood 115, 5111-5120 (2010).
D. Liu, J. Aguirre-Ghiso, Y. D. Estrada, L. Ossowski, EGFR is a transducer of the urokinase receptor initiated signal that is required for in vivo growth of a human carcinoma. Cancer Cell 1, 445-457 (2002).
J. Kiyan, R. Kiyan, H. Haller, I. Dumler, Urokinase-induced signaling in human vascular smooth muscle cells is mediated by PDGFR-β. EMBO J. 24, 1787-1797 (2005).
C.-H. Tang, Y. Wei, The urokinase receptor and integrins in cancer progression. Cell. Mol. Life Sci. 65, 1916-1932 (2008).
M. C. Kugler, Y. Wei, H. A. Chapman, Urokinase receptor and integrin interactions. Curr. Pharm. Des. 9, 1565-1574 (2003).
B. Degryse, M. Resnati, R.-P. Czekay, D. J. Loskutoff, F. Blasi, Domain 2 of the urokinase receptor contains an integrin-interacting epitope with intrinsic signaling activity: Generation of a new integrin inhibitor. J. Biol. Chem. 280, 24792-24803 (2005).
M. V. Carriero, S. Del Vecchio, M. Capozzoli, P. Franco, L. Fontana, A. Zannetti, G. Botti, G. D'Aiuto, M. Salvatore, M. P. Stoppelli, Urokinase receptor interacts with avb5 vitronectin receptor, promoting urokinase-dependent cell migration in breast cancer. Cancer Res. 59, 5307-5314 (1999).
Y. Wei, J. A. Eble, Z. Wang, J. A. Kreidberg, H. A. Chapman, Urokinase receptors promote β1 integrin function through interactions with integrin α3β1. Mol. Biol. Cell 12, 2975-2986 (2001).
A. Nykjaer, C. M. Petersen, B. Møller, P. H. Jensen, S. K. Moestrup, T. L. Holtet, M. Etzerodt, H. C. Thøgersen, M. Munch, P. A. Andreasen, Purified a2-macroglobulin receptor/LDL receptor-related protein binds urokinase.plasminogen activator inhibitor type-1 complex. Evidence that the a2-macroglobulin receptor mediates cellular degradation of urokinase receptor-bound complexes. J. Biol. Chem. 267, 14543-14546 (1992).
R.-P. Czekay, T. A. Kuemmel, R. A. Orlando, M. G. Farquhar, Direct binding of occupied urokinase receptor (uPAR) to LDL receptor-related protein is required for endocytosis of uPAR and regulation of cell surface urokinase activity. Mol. Biol. Cell 12, 1467-1479 (2001).
K. Cortese, M. Sahores, C. D. Madsen, C. Tacchetti, F. Blasi, Clathrin and LRP-1- independent constitutive endocytosis and recycling of uPAR. PLOS One 3, e3730 (2008).
F. Blasi, P. Carmeliet, uPAR: A versatile signalling orchestrator. Nat. Rev. Mol. Cell Biol. 3, 932-943 (2002).
L. Ossowski, J. A. Aguirre-Ghiso, Urokinase receptor and integrin partnership: Coordination of signaling for cell adhesion, migration and growth. Curr. Opin. Cell Biol. 12, 613-620 (2000).
G. W. Prager, J. Mihaly, P. M. Brunner, Y. Koshelnick, G. Hoyer-Hansen, B. R. Binder, Urokinase mediates endothelial cell survival via induction of the X-linked inhibitor of apoptosis protein. Blood 113, 1383-1390 (2009).
T. T. Chen, A. Luque, S. Lee, S. M. Anderson, T. Segura, M. L. Iruela-Arispe, Anchorage of VEGF to the extracellular matrix conveys differential signaling responses to endothelial cells. J. Cell Biol. 188, 595-609 (2010).
S. Sawamiphak, S. Seidel, C. L. Essmann, G. A. Wilkinson, M. E. Pitulescu, T. Acker, A. Acker-Palmer, Ephrin-B2 regulates VEGFR2 function in developmental and tumour angiogenesis. Nature 465, 487-491 (2010).
T. Jiang, J. Zhuang, H. Duan, Y. Luo, Q. Zeng, K. Fan, H. Yan, D. Lu, Z. Ye, J. Hao, J. Feng, D. Yang, X. Yan, CD146 is a coreceptor for VEGFR-2 in tumor angiogenesis. Blood 120, 2330-2339 (2012).
M. Nakayama, A. Nakayama, M. van Lessen, H. Yamamoto, S. Hoffmann, H. C. A. Drexler, N. Itoh, T. Hirose, G. Breier, D. Vestweber, J. A. Cooper, S. Ohno, K. Kaibuchi, R. H. Adams, Spatial regulation of VEGF receptor endocytosis in angiogenesis. Nat. Cell Biol. 15, 249-260 (2013).
Y. Wei, C.-H. Tang, Y. Kim, L. Robillard, F. Zhang, M. C. Kugler, H. A. Chapman, Urokinase receptors are required for α5β1 integrin-mediated signaling in tumor cells. J. Biol. Chem. 282, 3929-3939 (2007).
P. Chaurasia, J. A. Aguirre-Ghiso, O. D. Liang, H. Gardsvoll, M. Ploug, L. Ossowski, A region in urokinase plasminogen receptor domain III controlling a functional association with a5b1 integrin and tumor growth. J. Biol. Chem. 281, 14852-14863 (2006).
X. Z. West, N. Meller, N. L. Malinin, L. Deshmukh, J. Meller, G. H. Mahabeleshwar, M. E. Weber, B. A. Kerr, O. Vinogradova, T. V. Byzova, Integrin b3 crosstalk with VEGFR accommodating tyrosine phosphorylation as a regulatory switch. PLOS One 7, e31071 (2012).
S. Lakshmikanthan, M. Sobczak, C. Chun, A. Henschel, J. Dargatz, R. Ramchandran, M. Chrzanowska-Wodnicka, Rap1 promotes VEGFR2 activation and angiogenesis by a mechanism involving integrin ανβ3. Blood 118, 2015-2026 (2011).
Z. Liu, K. Kobayashi, M. van Dinther, S. H. van Heiningen, G. Valdimarsdottir, T. van Laar, M. Scharpfenecker, C. W. Lowik, M. J. Goumans, P. Ten Dijke, E. Pardali, VEGF and inhibitors of TGFβ type-I receptor kinase synergistically promote bloodvessel formation by inducing α5-integrin expression. J. Cell Sci. 122, 3294-3302 (2009).
E. Borges, Y. Jan, E. Ruoslahti, Platelet-derived growth factor receptor b and vascular endothelial growth factor receptor 2 bind to the β3 integrin through its extracellular domain. J. Biol. Chem. 275, 39867-39873 (2000).
S. Dimmeler, I. Fleming, B. Fisslthaler, C. Hermann, R. Busse, A. M. Zeiher, Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature 399, 601-605 (1999).
H. Gerhardt, M. Golding, M. Fruttiger, C. Ruhrberg, A. Lundkvist, A. Abramsson, M. Jeltsch, C. Mitchell, K. Alitalo, D. Shima, C. Betsholtz, VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia. J. Cell Biol. 161, 1163-1177 (2003).
G. A. McMahon, E. Petitclerc, S. Stefansson, E. Smith, M. K. K. Wong, R. J. Westrick, D. Ginsburg, P. C. Brooks, D. A. Lawrence, Plasminogen activator inhibitor-1 regulates tumor growth and angiogenesis. J. Biol. Chem. 276, 33964-33968 (2001).
M. Conese, A. Nykjaer, C. M. Petersen, O. Cremona, R. Pardi, P. A. Andreasen, J. Gliemann, E. I. Christensen, F. Blasi, Alpha-2 macroglobulin receptor/Ldl receptorrelated protein(Lrp)-dependent internalization of the urokinase receptor. J. Cell Biol. 131, 1609-1622 (1995).
E. Macia, M. Ehrlich, R. Massol, E. Boucrot, C. Brunner, T. Kirchhausen, Dynasore, a cell-permeable inhibitor of dynamin. Dev. Cell 10, 839-850 (2006).
T. H. Bugge, T. T. Suh, M. J. Flick, C. C. Daugherty, J. Romer, H. Solberg, V. Ellis, K. Danø, J. L. Degen, The receptor for urokinase-type plasminogen activator is not essential for mouse development or fertility. J. Biol. Chem. 270, 16886-16894 (1995).
M. Manetti, I. Rosa, M. Fazi, S. Guiducci, P. Carmeliet, L. Ibba-Manneschi, M. Matucci-Cerinic, Systemic sclerosis-like histopathological features in the myocardium of uPAR-deficient mice. Ann. Rheum. Dis. 10.1136/annrheumdis-2015-207803 (2015).
M. Manetti, I. Rosa, A. F. Milia, S. Guiducci, P. Carmeliet, L. Ibba-Manneschi, M. Matucci-Cerinic, Inactivation of urokinase-type plasminogen activator receptor (uPAR) gene induces dermal and pulmonary fibrosis and peripheral microvasculopathy in mice: A new model of experimental scleroderma? Ann. Rheum. Dis. 73, 1700-1709 (2014).
T. Tarui, N. Akakura, M. Majumdar, N. Andronicos, J. Takagi, A. P. Mazar, K. Bdeir, A. Kuo, S. V. Yarovoi, D. B. Cines, Y. Takada, Direct interaction of the kringle domain of urokinase-type plasminogen activator (uPA) and integrin avb3 induces signal transduction and enhances plasminogen activation. Thromb. Haemost. 95, 524-534 (2006).
Y. Takada, Potential role of kringle-integrin interaction in plasmin and uPA actions (a hypothesis). J. Biomed. Biotechnol. 2012, 136302 (2012).
Y. Liu, R. Pixley, M. Fusaro, G. Godoy, E. Kim, M. E. Bromberg, R. W. Colman, Cleaved high-molecular-weight kininogen and its domain 5 inhibit migration and invasion of human prostate cancer cells through the epidermal growth factor receptor pathway. Oncogene 28, 2756-2765 (2009).
L. Fang, S.-H. Choi, J. S. Baek, C. Liu, F. Almazan, F. Ulrich, P. Wiesner, A. Taleb, E. Deer, J. Pattison, J. Torres-Vázquez, A. C. Li, Y. I. Miller, Control of angiogenesis by AIBP-mediated cholesterol efflux. Nature 498, 118-122 (2013).
S. Ikeda, M. Ushio-Fukai, L. Zuo, T. Tojo, S. Dikalov, N. A. Patrushev, R. W. Alexander, Novel role of ARF6 in vascular endothelial growth factor-induced signaling and angiogenesis. Circ. Res. 96, 467-475 (2005).
L. Labrecque, I. Royal, D. S. Surprenant, C. Patterson, D. Gingras, R. Béliveau, Regulation of vascular endothelial growth factor receptor-2 activity by caveolin-1 and plasma membrane cholesterol. Mol. Biol. Cell 14, 334-347 (2003).
W.-x. Liao, L. Feng, H. Zhang, J. Zheng, T. R. Moore, D.-b. Chen, Compartmentalizing VEGF-induced ERK2/1 signaling in placental artery endothelial cell caveolae: A paradoxical role of caveolin-1 in placental angiogenesis in vitro. Mol. Endocrinol. 23, 1428-1444 (2009).
A. Noghero, A. Perino, G. Seano, E. Saglio, G. Lo Sasso, F. Veglio, L. Primo, E. Hirsch, F. Bussolino, F. Morello, Liver X receptor activation reduces angiogenesis by impairing lipid raft localization and signaling of vascular endothelial growth factor receptor-2. Arterioscler. Thromb. Vasc. Biol. 32, 2280-2288 (2012).
A. Tiwari, J. J. Jung, S. M. Inamdar, D. Nihalani, A. Choudhury, The myosin motor Myo1c is required for VEGFR2 delivery to the cell surface and for angiogenic signaling. Am. J. Physiol. Heart Circ. Physiol. 304, H687-H696 (2013).
F. Shi, J. Sottile, Caveolin-1-dependent b1 integrin endocytosis is a critical regulator of fibronectin turnover. J. Cell Sci. 121, 2360-2371 (2008).
F. Margheri, A. Chillà, A. Laurenzana, S. Serratì, B. Mazzanti, R. Saccardi, M. Santosuosso, G. Danza, N. Sturli, F. Rosati, L. Magnelli, L. Papucci, L. Calorini, F. Bianchini, M. Del Rosso, G. Fibbi, Endothelial progenitor cell-dependent angiogenesis requires localization of the full-length form of uPAR in caveolae. Blood 118, 3743-3755 (2011).
A. Nykjær, M. Conese, E. I. Christensen, D. Olson, O. Cremona, J. Gliemann, F. Blasi, Recycling of the urokinase receptor upon internalization of the uPA: Serpin complexes. EMBO J. 16, 2610-2620 (1997).
E. A. Jaffe, R. L. Nachman, C. G. Becker, C. R. Minick, Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J. Clin. Invest. 52, 2745-2756 (1973).
M. Dewerchin, A. V. Nuffelen, G. Wallays, A. Bouché, L. Moons, P. Carmeliet, R. C. Mulligan, D. Collen, Generation and characterization of urokinase receptor-deficient mice. J. Clin. Invest. 97, 870-878 (1996).
A. Passaniti, R. M. Taylor, R. Pili, Y. Guo, P. V. Long, J. A. Haney, R. R. Pauly, D. S. Grant, G. R. Martin, A simple, quantitative method for assessing angiogenesis and antiangiogenic agents using reconstituted basement membrane, heparin, and FGF. Lab. Invest. 67, 519-528 (1992).