TY - JOUR
T1 - VEGFR-3 controls tip to stalk conversion at vessel fusion sites by reinforcing Notch signalling
AU - Tammela, Tuomas
AU - Zarkada, Georgia
AU - Nurmi, Harri
AU - Jakobsson, Lars
AU - Heinolainen, Krista
AU - Tvorogov, Denis
AU - Zheng, Wei
AU - Franco, Claudio A.
AU - Murtomäki, Aino
AU - Aranda, Evelyn
AU - Miura, Naoyuki
AU - Ylä-Herttuala, Seppo
AU - Fruttiger, Marcus
AU - Mäkinen, Taija
AU - Eichmann, Anne
AU - Pollard, Jeffrey W.
AU - Gerhardt, Holger
AU - Alitalo, Kari
N1 - Funding Information:
We would like to thank T. Petrova (CePO, CHUV and University of Lausanne, Switzerland) for the Foxc2+/− mice, M. Achen and S. Stacker (Peter MacCallum Cancer Centre, Melbourne, Australia) for the Vegfd−/− mice, B. Pytowski at Eli Lilly for VEGFR-2-and VEGFR-3-blocking antibodies, M. Jeltsch (Molecular/Cancer Biology Laboratory, University of Helsinki, Finland) for generating VEGF-C antibodies, S. Kaijalainen (Molecular/Cancer Biology Laboratory, University of Helsinki, Finland) for generating mDll4-Fc and mDll4–ECTM–eGFP expression vectors, A. Alitalo (Institute of Pharmaceutical Sciences, ETH Zurich, Switzerland) for valuable help with experiments and K. Helenius for critical comments on the manuscript. The Biomedicum Molecular Imaging Unit is acknowledged for microscopy services, and N. Ihalainen, T. Laakkonen, K. Salo and T. Tainola for excellent technical assistance, as well as personnel of the Meilahti Experimental Animal Center (University of Helsinki) for expert animal husbandry. We also thank I. Rosewell (London Research Institute, UK) for generation of chimaeric mice. This work was supported by grants from the Academy of Finland, the Association for International Cancer Research, the Finnish Cancer Organizations, the Helsinki University Research Fund, the Sigrid Juselius Foundation, the Louis-Jeantet Foundation and the European Research Council (ERC-2010-AdG-268804-TX-FACTORS). T.T. was supported by personal grants from the Emil Aaltonen Foundation, the K. Albin Johansson Foundation, the Finnish Medical Foundation, the Maud Kuistila Foundation, the Orion-Farmos Research Foundation and the Paulo Foundation. G.Z. was supported by personal grants from the K. Albin Johansson Foundation, the Finnish Medical Foundation, The Paulo Foundation, the Ida Montin Foundation and the Orion-Farmos Research Foundation. H.G. was supported by Cancer Research UK, the Lister Institute of Preventive Medicine, the European Molecular Biology Organization (EMBO) Young Investigator Programme and the Leducq Transatlantic Network ARTEMIS. L.J. was supported by an EMBO long-term postdoctoral fellowship. C.A.F. was supported by a Marie Curie FP7 postdoctoral fellowship.
PY - 2011/10
Y1 - 2011/10
N2 - Angiogenesis, the growth of new blood vessels, involves specification of endothelial cells to tip cells and stalk cells, which is controlled by Notch signalling, whereas vascular endothelial growth factor receptor (VEGFR)-2 and VEGFR-3 have been implicated in angiogenic sprouting. Surprisingly, we found that endothelial deletion of Vegfr3, but not VEGFR-3-blocking antibodies, postnatally led to excessive angiogenic sprouting and branching, and decreased the level of Notch signalling, indicating that VEGFR-3 possesses passive and active signalling modalities. Furthermore, macrophages expressing the VEGFR-3 and VEGFR-2 ligand VEGF-C localized to vessel branch points, and Vegfc heterozygous mice exhibited inefficient angiogenesis characterized by decreased vascular branching. FoxC2 is a known regulator of Notch ligand and target gene expression, and Foxc2 +/-;Vegfr3 +/-' compound heterozygosity recapitulated homozygous loss of Vegfr3. These results indicate that macrophage-derived VEGF-C activates VEGFR-3 in tip cells to reinforce Notch signalling, which contributes to the phenotypic conversion of endothelial cells at fusion points of vessel sprouts.
AB - Angiogenesis, the growth of new blood vessels, involves specification of endothelial cells to tip cells and stalk cells, which is controlled by Notch signalling, whereas vascular endothelial growth factor receptor (VEGFR)-2 and VEGFR-3 have been implicated in angiogenic sprouting. Surprisingly, we found that endothelial deletion of Vegfr3, but not VEGFR-3-blocking antibodies, postnatally led to excessive angiogenic sprouting and branching, and decreased the level of Notch signalling, indicating that VEGFR-3 possesses passive and active signalling modalities. Furthermore, macrophages expressing the VEGFR-3 and VEGFR-2 ligand VEGF-C localized to vessel branch points, and Vegfc heterozygous mice exhibited inefficient angiogenesis characterized by decreased vascular branching. FoxC2 is a known regulator of Notch ligand and target gene expression, and Foxc2 +/-;Vegfr3 +/-' compound heterozygosity recapitulated homozygous loss of Vegfr3. These results indicate that macrophage-derived VEGF-C activates VEGFR-3 in tip cells to reinforce Notch signalling, which contributes to the phenotypic conversion of endothelial cells at fusion points of vessel sprouts.
UR - http://www.scopus.com/inward/record.url?scp=80053564674&partnerID=8YFLogxK
U2 - 10.1038/ncb2331
DO - 10.1038/ncb2331
M3 - Article
C2 - 21909098
SN - 1465-7392
VL - 13
SP - 1202
EP - 1213
JO - Nature Cell Biology
JF - Nature Cell Biology
IS - 10
ER -