Inhibition of Dll4 signalling inhibits tumour growth by deregulating angiogenesis

Abstract

Haploinsufficiency of Dll4, a vascular-specific Notch ligand, has shown that it is essential for embryonic vascular development and arteriogenesis1,2,3. Mechanistically, it is unclear how the Dll4-mediated Notch pathway contributes to complex vascular processes that demand meticulous coordination of multiple signalling pathways. Here we show that Dll4-mediated Notch signalling has a unique role in regulating endothelial cell proliferation and differentiation. Neutralizing Dll4 with a Dll4-selective antibody rendered endothelial cells hyperproliferative, and caused defective cell fate specification or differentiation both in vitro and in vivo. In addition, blocking Dll4 inhibited tumour growth in several tumour models. Remarkably, antibodies against Dll4 and antibodies against vascular endothelial growth factor (VEGF) had paradoxically distinct effects on tumour vasculature. Our data also indicate that Dll4-mediated Notch signalling is crucial during active vascularization, but less important for normal vessel maintenance. Furthermore, unlike blocking Notch signalling globally, neutralizing Dll4 had no discernable impact on intestinal goblet cell differentiation4,5, supporting the idea that Dll4-mediated Notch signalling is largely restricted to the vascular compartment. Therefore, targeting Dll4 might represent a broadly efficacious and well-tolerated approach for the treatment of solid tumours.

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Figure 1: Dll4-mediated Notch signalling regulates endothelial cell proliferation.
Figure 2: Dll4-mediated Notch signalling regulates endothelial cell differentiation.
Figure 3: Selective blocking of Dll4 disrupts tumour angiogenesis and inhibits tumour growth.
Figure 4: Dll4/Notch in the homeostasis of mouse small intestine.

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Acknowledgements

We thank G. Fuh and V. Lee for the antibody phage libraries, L. DeGuzman and J. Zavala-Solorio for technical assistance, M. Vasser and P. Ng for oligonucleotide synthesis and purification, Genentech Protein Chemistry, Pathology Lab and core DNA sequencing facility for support services, W. Ye, C. Siebel, N. Ferrara, J. Zha and K. O'Rourke for discussion, and V. Dixit for encouragement.

Author Contributions J. R. and G. Z. designed and performed experiments. Y. W. supervised antibody generation. S. S. and W. L generated antibodies. Y. C., J. K. and R. J. W. performed neonatal mouse retina studies and data analysis. C. C. performed pathological analysis. I. K. provided expertise and assistance on confocal imaging. M. S. and M. C. performed mouse intestine studies. C. T. and J. H. generated analytical reagents. F. de S. supervised mouse intestine studies. G. P. supervised tumour studies of combination therapy. M. Y. designed and conceptualized the study, and wrote the manuscript. All authors made comments on the manuscript.

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Correspondence to Minhong Yan.

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Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

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Ridgway, J., Zhang, G., Wu, Y. et al. Inhibition of Dll4 signalling inhibits tumour growth by deregulating angiogenesis. Nature 444, 1083–1087 (2006). https://doi.org/10.1038/nature05313

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