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Blockade of Dll4 inhibits tumour growth by promoting non-productive angiogenesis


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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Figure 1: Dll4 is expressed in tumour vessels, and its expression is dependent on VEGF signalling.
Figure 2: Blockade of Dll4/Notch signalling results in smaller C6 tumours with increased vessel density.
Figure 3: Despite an increase in blood vessel density, tumours overexpressing Dll4–Fc have increased hypoxia and poor vascular perfusion.
Figure 4: Systemic delivery of Dll4–Fc using adenovirus results in smaller C6 tumours and increased vessel density, similar to effects of local tumour overexpression.
Figure 5: Systemic delivery of Dll4–Fc or blocking Dll4 antibodies to mice bearing tumours that are resistant to blockade of VEGF results in decreased tumour growth and dramatic changes in tumour vessels.


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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.

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Correspondence to Gavin Thurston.

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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).

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