NOTCH Decoys That Selectively Block DLL/NOTCH or JAG/NOTCH Disrupt Angiogenesis by Unique Mechanisms to Inhibit Tumor Growth

A proangiogenic role for Jagged (JAG)-dependent activation of NOTCH signaling in the endothelium has yet to be described. Using proteins that encoded different NOTCH1 EGF-like repeats, we identified unique regions of Delta-like ligand (DLL)–class and JAG-class ligand–receptor interactions, and developed NOTCH decoys that function as ligand-specific NOTCH inhibitors. N110–24 decoy blocked JAG1/JAG2–mediated NOTCH1 signaling, angiogenic sprouting in vitro, and retinal angiogenesis, demonstrating that JAG-dependent NOTCH signal activation promotes angiogenesis. In tumors, N110–24 decoy reduced angiogenic sprouting, vessel perfusion, pericyte coverage, and tumor growth. JAG–NOTCH signaling uniquely inhibited expression of antiangiogenic soluble (s) VEGFR1/sFLT1. N11–13 decoy interfered with DLL1–DLL4-mediated NOTCH1 signaling and caused endothelial hypersprouting in vitro, in retinal angiogenesis, and in tumors. Thus, blockade of JAG- or DLL-mediated NOTCH signaling inhibits angiogenesis by distinct mechanisms. JAG–NOTCH signaling positively regulates angiogenesis by suppressing sVEGFR1–sFLT1 and promoting mural–endothelial cell interactions. Blockade of JAG-class ligands represents a novel, viable therapeutic approach to block tumor angiogenesis and growth. Significance: This is the first report identifying unique regions of the NOTCH1 extracellular domain that interact with JAG-class and DLL-class ligands. Using this knowledge, we developed therapeutic agents that block JAG-dependent NOTCH signaling and demonstrate for the first time that JAG blockade inhibits experimental tumor growth by targeting tumor angiogenesis. Cancer Discov; 5(2); 182–97. ©2014 AACR. See related commentary by Briot and Iruela-Arispe, p. 112 This article is highlighted in the In This Issue feature, p. 97