Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis (original) (raw)
- Letter
- Published: 28 January 2007
- Li-Kun Phng4,
- Jennifer J. Hofmann5,
- Elisabet Wallgard1,2,
- Leigh Coultas6,
- Per Lindblom4 nAff8,
- Jackelyn Alva5,
- Ann-Katrin Nilsson1 nAff8,
- Linda Karlsson1 nAff8,
- Nicholas Gaiano7,
- Keejung Yoon7,
- Janet Rossant6,
- M. Luisa Iruela-Arispe5,
- Mattias Kalén1,2 na1,
- Holger Gerhardt4 na1 &
- …
- Christer Betsholtz2,3 na1
Nature volume 445, pages 776–780 (2007)Cite this article
- 16k Accesses
- 1313 Citations
- 13 Altmetric
- Metrics details
Abstract
In sprouting angiogenesis, specialized endothelial tip cells lead the outgrowth of blood-vessel sprouts towards gradients of vascular endothelial growth factor (VEGF)-A1,2. VEGF-A is also essential for the induction of endothelial tip cells2, but it is not known how single tip cells are selected to lead each vessel sprout, and how tip-cell numbers are determined. Here we present evidence that delta-like 4 (Dll4)–Notch1 signalling regulates the formation of appropriate numbers of tip cells to control vessel sprouting and branching in the mouse retina. We show that inhibition of Notch signalling using γ-secretase inhibitors, genetic inactivation of one allele of the endothelial Notch ligand Dll4, or endothelial-specific genetic deletion of Notch1, all promote increased numbers of tip cells. Conversely, activation of Notch by a soluble jagged1 peptide leads to fewer tip cells and vessel branches. Dll4 and reporters of Notch signalling are distributed in a mosaic pattern among endothelial cells of actively sprouting retinal vessels. At this location, Notch1-deleted endothelial cells preferentially assume tip-cell characteristics. Together, our results suggest that Dll4–Notch1 signalling between the endothelial cells within the angiogenic sprout serves to restrict tip-cell formation in response to VEGF, thereby establishing the adequate ratio between tip and stalk cells required for correct sprouting and branching patterns. This model offers an explanation for the dose-dependency and haploinsufficiency of the Dll4 gene3,4,5, and indicates that modulators of Dll4 or Notch signalling, such as γ-secretase inhibitors developed for Alzheimer’s disease, might find usage as pharmacological regulators of angiogenesis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Additional access options:
Similar content being viewed by others
References
- Ruhrberg, C. et al. Spatially restricted patterning cues provided by heparin-binding VEGF-A control blood vessel branching morphogenesis. Genes Dev. 16, 2684–2698 (2002)
Article CAS Google Scholar - Gerhardt, H. et al. VEGF guides angiogenic sprouting utilizing endothelial tip-cell filopodia. J. Cell Biol. 161, 1163–1177 (2003)
Article CAS Google Scholar - Gale, N. W. et al. Haploinsufficiency of delta-like 4 ligand results in embryonic lethality due to major defects in arterial and vascular development. Proc. Natl Acad. Sci. USA 101, 15949–15954 (2004)
Article ADS CAS Google Scholar - Krebs, L. T. et al. Haploinsufficienct lethality and formation of arteriovenous malformations in Notch pathway mutants. Genes Dev. 18, 2469–2473 (2004)
Article CAS Google Scholar - Duarte, A. et al. Dosage-sensitive requirement for mouse Dll4 in artery development. Genes Dev. 18, 2474–2478 (2004)
Article CAS Google Scholar - Uv, A., Cantera, R. & Samakovlis, C. Drosophila tracheal morphogenesis: intricate cellular solutions to basic plumbing problems. Trends Cell Biol. 13, 301–309 (2003)
Article CAS Google Scholar - Carmeliet, P. & Tessier-Lavigne, M. Common mechanisms of nerve and blood vessel wiring. Nature 436, 193–200 (2005)
Article ADS CAS Google Scholar - Lu, X. et al. The netrin receptor UNC5B mediates guidance events controlling morphogenesis of the vascular system. Nature 432, 179–186 (2004)
Article ADS CAS Google Scholar - Torres-Vazquez, J. et al. Semaphorin-plexin signaling guides patterning of the developing vasculature. Dev. Cell 7, 117–123 (2004)
Article CAS Google Scholar - Llimargas, M. The Notch pathway helps to pattern the tips of the Drosophila tracheal branches by selecting cell fates. Development 126, 2355–2364 (1999)
CAS PubMed Google Scholar - Steneberg, P., Hemphala, J. & Samakovlis, C. Dpp and Notch specify the fusion cell fate in the dorsal branches of the Drosophila trachea. Mech. Dev. 87, 153–163 (1999)
Article CAS Google Scholar - Ghabrial, A. S. & Krasnow, M. A. Social interactions among epithelial cells during tracheal branching morphogenesis. Nature 441, 746–749 (2006)
Article ADS CAS Google Scholar - Shawber, C. J. & Kitajewski, J. Notch function in the vasculature: Insights from zebrafish, mouse and man. Bioessays 26, 225–234 (2004)
Article CAS Google Scholar - Dovey, H. F. et al. Functional γ-secretase inhibitors reduce β-amyloid peptide levels in brain. J. Neurochem. 76, 173–181 (2001)
Article CAS Google Scholar - Searfoss, G. H. et al. Adipsin, a biomarker of gastrointestinal toxicity mediated by a functional γ-secretase inhibitor. J. Biol. Chem. 278, 46107–46116 (2003)
Article CAS Google Scholar - van Es, J. H. et al. Notch/γ-secretase inhibition turns proliferative cells in intestinal crypts and adenomas into goblet cells. Nature 435, 959–963 (2005)
Article ADS CAS Google Scholar - Kopan, R. & Ilagan, M. X. G. γ-secretases: proteasome of the membrane? Nature Rev. Mol. Cell Biol. 5, 499–504 (2004)
Article CAS Google Scholar - Claxton, S. & Fruttiger, M. Periodic Delta-like 4 expression in developing retinal arteries. Gene Expr. Patterns 5, 123–127 (2004)
Article CAS Google Scholar - Shutter, J. R. et al. Dll4, a novel Notch ligand expressed in arterial endothelium. Genes Dev. 14, 1313–1318 (2000)
CAS PubMed PubMed Central Google Scholar - Lawson, N. D. et al. Notch signaling is required for arterial-venous differentiation during embryonic vascular development. Development 128, 3675–3683 (2001)
CAS PubMed Google Scholar - Domenga, V. et al. Notch3 is required for arterial identity and maturation of vascular smooth muscle cells. Genes Dev. 18, 2730–2735 (2004)
Article CAS Google Scholar - Krebs, L. T. et al. Notch signaling is essential for vascular morphogenesis in mice. Genes Dev. 14, 1343–1352 (2000)
CAS PubMed PubMed Central Google Scholar - Limbourg, F. P. et al. Essential role of endothelial Notch1 in angiogenesis. Circulation 111, 1826–1832 (2005)
Article CAS Google Scholar - Monvoisin, A. et al. VE-cadherin-CreERT2 transgenic mouse: A model for inducible recombination in the endothelium. Dev. Dyn. 235, 3413–3422 (2006)
Article CAS Google Scholar - Wolfer, A. et al. Inactivation of Notch1 in immature thymocytes does not perturb CD4 or CD8 T cell development. Nature Immunol. 2, 235–241 (2001)
Article CAS Google Scholar - Duncan, A. W. et al. Integration of Notch and Wnt signaling in hematopoietic stem cell maintenance. Nature Immunol. 6, 314–322 (2005)
Article CAS Google Scholar - Itoh, M. et al. Mind Bomb is a ubiquitin ligase that is essential for efficient activation of Notch signaling by Delta. Dev. Cell 4, 67–82 (2003)
Article CAS Google Scholar - Weijzen, S. et al. The notch ligand Jagged-1 is able to induce maturation of monocyte-derived human dendritic cells. J. Immunol. 169, 4273–4278 (2002)
Article CAS Google Scholar - Grunstein, J., Masbad, J. J., Hickey, R., Giordano, F. & Johnson, R. S. Isoforms of vascular endothelial growth factor act in a coordinate fashion to recruit and expand tumor vasculature. Mol. Cell. Biol. 20, 7282–7291 (2000)
Article CAS Google Scholar - Ferrara, N. Vascular endothelial growth factor: Basic science and clinical progress. Endocr. Rev. 25, 581–611 (2004)
Article CAS Google Scholar
Acknowledgements
We thank F. Radke for providing Notch1floxed/floxed mice. Support from the following foundations and granting agencies is acknowledged: Swedish Cancer Society, Association for International Cancer Research, European Union, the Novo Nordisk, Strategic Research, Söderberg, Hedlund, Wallenberg and Inga-Britt and Arne Lundberg Foundations (to C.B.); National Institutes of Health (US, NIH) and JH (USPHS National Research Service Award) (to L.I.-A.). H.G., L.-K.P. and P.L. are supported by Cancer Research UK. We acknowledge the Swegene Centre for Cellular Imaging at Gothenburg University for the use of imaging equipment, and the Light Microscopy Service and Peptide Synthesis Laboratory, London Research Institute (Cancer Research UK) for technical assistance.
Author information
Author notes
- Per Lindblom, Ann-Katrin Nilsson & Linda Karlsson
Present address: Present addresses: Molecular Toxicology, Safety Assessment, AstraZeneca R&D, SE-151 85 Södertälje, Sweden (P.L.); Stem Cell Center, BMC B10, Klinikg. 26, Lund University, SE-221 84 Lund, Sweden (A.-K.N.); Department of Physiology, Göteborg University, P.O. Box 434, SE-405 30 Göteborg, Sweden (L.K.)., - Mattias Kalén, Holger Gerhardt and Christer Betsholtz: These authors contributed equally to this work.
Authors and Affiliations
- AngioGenetics Sweden AB, Scheeles väg 2, SE-171 77 Stockholm, Sweden,
Mats Hellström, Elisabet Wallgard, Ann-Katrin Nilsson, Linda Karlsson & Mattias Kalén - Department of Medical Biochemistry and Biophysics, Division of Matrix Biology, and,
Mats Hellström, Elisabet Wallgard, Mattias Kalén & Christer Betsholtz - Department of Medicine, Karolinska Institutet, SE 171 77 Stockholm, Sweden,
Christer Betsholtz - Vascular Biology Laboratory, London Research Institute, Cancer Research UK, London WC2A 3PX, UK,
Li-Kun Phng, Per Lindblom & Holger Gerhardt - Department of Molecular, Cell and Developmental Biology and Molecular Biology Institute, UCLA, Los Angeles, California 90095, USA,
Jennifer J. Hofmann, Jackelyn Alva & M. Luisa Iruela-Arispe - Program for Developmental Biology, Hospital for Sick Children, Toronto, Ontario M5G 1X8, Canada,
Leigh Coultas & Janet Rossant - Neurology, Neuroscience and Oncology, Institute for Cell Engineering, Johns Hopkins University, Baltimore, Maryland 21205, USA,
Nicholas Gaiano & Keejung Yoon
Authors
- Mats Hellström
You can also search for this author inPubMed Google Scholar - Li-Kun Phng
You can also search for this author inPubMed Google Scholar - Jennifer J. Hofmann
You can also search for this author inPubMed Google Scholar - Elisabet Wallgard
You can also search for this author inPubMed Google Scholar - Leigh Coultas
You can also search for this author inPubMed Google Scholar - Per Lindblom
You can also search for this author inPubMed Google Scholar - Jackelyn Alva
You can also search for this author inPubMed Google Scholar - Ann-Katrin Nilsson
You can also search for this author inPubMed Google Scholar - Linda Karlsson
You can also search for this author inPubMed Google Scholar - Nicholas Gaiano
You can also search for this author inPubMed Google Scholar - Keejung Yoon
You can also search for this author inPubMed Google Scholar - Janet Rossant
You can also search for this author inPubMed Google Scholar - M. Luisa Iruela-Arispe
You can also search for this author inPubMed Google Scholar - Mattias Kalén
You can also search for this author inPubMed Google Scholar - Holger Gerhardt
You can also search for this author inPubMed Google Scholar - Christer Betsholtz
You can also search for this author inPubMed Google Scholar
Corresponding authors
Correspondence toMats Hellström or Holger Gerhardt.
Ethics declarations
Competing interests
[Competing Interests Statement: M.H. and M.K. are employed by AngioGenetics Sweden AB. C.B. receives funding from and is a consultant of AngioGenetics Sweden AB.]
Supplementary information
Supplementary Information
This file contains Supplementary Methods, Supplementary Figures S1-S11 with Legends and additional references. (PDF 7910 kb)
Rights and permissions
About this article
Cite this article
Hellström, M., Phng, LK., Hofmann, J. et al. Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis.Nature 445, 776–780 (2007). https://doi.org/10.1038/nature05571
- Received: 16 September 2006
- Accepted: 05 January 2007
- Published: 28 January 2007
- Issue Date: 15 February 2007
- DOI: https://doi.org/10.1038/nature05571