Tumour growth requires accompanying expansion of the host vasculature, with tumour progression often correlated with vascular density. Vascular endothelial growth factor (VEGF) is the best-characterized inducer of tumour angiogenesis. We report that VEGF dynamically regulates tumour endothelial expression of Delta-like ligand 4 (Dll4), which was previously shown to be absolutely required for normal embryonic vascular development. To define Dll4 function in tumour angiogenesis, we manipulated this pathway in murine tumour models using several approaches. Here we show that blockade resulted in markedly increased tumour vascularity, associated with enhanced angiogenic sprouting and branching. Paradoxically, this increased vascularity was non-productive—as shown by poor perfusion and increased hypoxia, and most importantly, by decreased tumour growth—even for tumours resistant to anti-VEGF therapy. Thus, VEGF-induced Dll4 acts as a negative regulator of tumour angiogenesis; its blockade results in a striking uncoupling of tumour growth from vessel density, presenting a novel therapeutic approach even for tumours resistant to anti-VEGF therapies.
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Folkman, J. The role of angiogenesis in tumor growth. Semin. Cancer Biol. 3, 65–71 (1992)
Ferrara, N. Vascular endothelial growth factor as a target for anticancer therapy. Oncologist 9, (Suppl. 1)2–10 (2004)
Rudge, J. S. et al. VEGF trap as a novel antiangiogenic treatment currently in clinical trials for cancer and eye diseases, and VelociGene-based discovery of the next generation of angiogenesis targets. Cold Spring Harb. Symp. Quant. Biol. 70, 411–418 (2005)
Holash, J. et al. VEGF-Trap: a VEGF blocker with potent antitumor effects. Proc. Natl Acad. Sci. USA 99, 11393–11398 (2002)
Hurwitz, H. et al. Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N. Engl. J. Med. 350, 2335–2342 (2004)
Laskin, J. J. & Sandler, A. B. First-line treatment for advanced non-small-cell lung cancer. Oncology 19, 1671–6; discussion 1678–80. (2005)
Casanovas, O., Hicklin, D. J., Bergers, G. & Hanahan, D. Drug resistance by evasion of antiangiogenic targeting of VEGF signaling in late-stage pancreatic islet tumors. Cancer Cell 8, 299–309 (2005)
Jain, R. K., Duda, D. G., Clark, J. W. & Loeffler, J. S. Lessons from phase III clinical trials on anti-VEGF therapy for cancer. Nature Clin. Pract. Oncol. 3, 24–40 (2006)
Kerbel, R. S. et al. Possible mechanisms of acquired resistance to anti-angiogenic drugs: implications for the use of combination therapy approaches. Cancer Metastasis Rev. 20, 79–86 (2001)
Yancopoulos, G. D. et al. Vascular-specific growth factors and blood vessel formation. Nature 407, 242–248 (2000)
Jain, R. K. Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science 307, 58–62 (2005)
Carmeliet, P. Angiogenesis in life, disease and medicine. Nature 438, 932–936 (2005)
Shawber, C. J. & Kitajewski, J. Notch function in the vasculature: insights from zebrafish, mouse and man. Bioessays 26, 225–234 (2004)
Artavanis-Tsakonas, S., Rand, M. D. & Lake, R. J. Notch signaling: cell fate control and signal integration in development. Science 284, 770–776 (1999)
Gridley, T. Notch signaling during vascular development. Proc. Natl Acad. Sci. USA 98, 5377–5378 (2001)
Duarte, A. et al. Dosage-sensitive requirement for mouse Dll4 in artery development. Genes Dev. 18, 2474–2478 (2004)
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)
Krebs, L. T. et al. Haploinsufficient lethality and formation of arteriovenous malformations in Notch pathway mutants. Genes Dev. 18, 2469–2473 (2004)
Mailhos, C. et al. Delta4, an endothelial specific notch ligand expressed at sites of physiological and tumor angiogenesis. Differentiation 69, 135–144 (2001)
Patel, N. S. et al. Up-regulation of delta-like 4 ligand in human tumor vasculature and the role of basal expression in endothelial cell function. Cancer Res. 65, 8690–8697 (2005)
Hicks, C. et al. A secreted Delta1–Fc fusion protein functions both as an activator and inhibitor of Notch1 signaling. J. Neurosci. Res. 68, 655–667 (2002)
Taylor, K. L., Henderson, A. M. & Hughes, C. C. Notch activation during endothelial cell network formation in vitro targets the basic HLH transcription factor HESR-1 and downregulates VEGFR-2/KDR expression. Microvasc. Res. 64, 372–383 (2002)
Iso, T., Kedes, L. & Hamamori, Y. HES and HERP families: multiple effectors of the Notch signaling pathway. J. Cell. Physiol. 194, 237–255 (2003)
Shawber, C. J., Das, I., Francisco, E. & Kitajewski, J. Notch signaling in primary endothelial cells. Ann. NY Acad. Sci. 995, 162–170 (2003)
Karsan, A. The role of notch in modeling and maintaining the vasculature. Can. J. Physiol. Pharmacol. 83, 14–23 (2005)
Lamar, E. et al. Nrarp is a novel intracellular component of the Notch signaling pathway. Genes Dev. 15, 1885–1899 (2001)
Krebs, L. T., Deftos, M. L., Bevan, M. J. & Gridley, T. The Nrarp gene encodes an ankyrin-repeat protein that is transcriptionally regulated by the Notch signaling pathway. Dev. Biol. 238, 110–119 (2001)
Krneta, J. et al. Dissociation of angiogenesis and tumorigenesis in follistatin- and activin-expressing tumors. Cancer Res. 66, 5686–5695 (2006)
Lee, C. G. et al. Anti-vascular endothelial growth factor treatment augments tumor radiation response under normoxic or hypoxic conditions. Cancer Res. 60, 5565–5570 (2000)
Jain, R. K. Tumor angiogenesis and accessibility: role of vascular endothelial growth factor. Semin. Oncol. 29, 3–9 (2002)
Jain, R. K. Antiangiogenic therapy for cancer: current and emerging concepts. Oncology. 19, 7–16 (2005)
Valenzuela, D. M. et al. High-throughput engineering of the mouse genome coupled with high-resolution expression analysis. Nature Biotechnol. 21, 652–659 (2003)
Thurston, G., Baluk, P., Hirata, A. & McDonald, D. M. Permeability-related changes revealed at endothelial cell borders in inflamed venules by lectin binding. Am. J. Physiol. 271, H2547–H2562 (1996)
Liu, Z. J. et al. Inhibition of endothelial cell proliferation by Notch1 signaling is mediated by repressing MAPK and PI3K/Akt pathways and requires MAML1. FASEB J. 20, 1009–1011 (2006)
We acknowledge the following Regeneron colleagues: Y. Wei for gene expression analysis, A. Adler, A. Rafique, B. Li, H. Huang, E. Pasnikowski, J. McClain, E. Burova, D. Hylton, P. Burfeind and J. Griffiths for technical assistance, S. Staton for assistance with graphics, and S. Wiegand, I. Lobov, T. Daly, S. Davis, E. Ioffe, J. Holash and J. Rudge for scientific input.
Author Contributions I. N.-T. directed and helped perform tumour experiments, generation of tumour lines, immunohistochemical staining, and data analysis. C.D. directed, helped perform, and analysed in vitro experiments. N.J.P. helped develop protein reagents and biochemical assays. S.C. performed and helped analyse tumour experiments and construction of tumour cell lines. P.B. performed and helped analyse immunohistochemical studies. N.W.G. helped perform and analyse experiments with gene-targeted mice. H.C.L. helped perform and analyse gene expression studies. G.D.Y. helped analyse and interpret results. G.T. helped design experiments, analyse data and interpret results.
All authors are employees of Regeneron Pharmaceuticals.
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Noguera-Troise, I., Daly, C., Papadopoulos, N. et al. Blockade of Dll4 inhibits tumour growth by promoting non-productive angiogenesis. Nature 444, 1032–1037 (2006). https://doi.org/10.1038/nature05355
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