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.


CONCLUSIONS
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.
Department of Obstetrics/Gynecology, Columbia University Medical Center, Columbia University, New York, New York. Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand.
Thaned
T
Kangsamaksin
Department of Obstetrics/Gynecology, Columbia University Medical Center, Columbia University, New York, New York. Division of Genetics, Department of Biosciences, Viikki Biocenter, University of Helsinki, Helsinki, Finland.
Aino
A
Murtomaki
Department of Obstetrics/Gynecology, Columbia University Medical Center, Columbia University, New York, New York.
Natalie M
NM
Kofler
Department of Obstetrics/Gynecology, Columbia University Medical Center, Columbia University, New York, New York.
Henar
H
Cuervo
Department of Obstetrics/Gynecology, Columbia University Medical Center, Columbia University, New York, New York.
Reyhaan A
RA
Chaudhri
Department of Obstetrics/Gynecology, Columbia University Medical Center, Columbia University, New York, New York.
Ian W
IW
Tattersall
Department of Pathology and Cellular Biology, Columbia University Medical Center, Columbia University, New York, New York.
Paul E
PE
Rosenstiel
Department of Obstetrics/Gynecology, Columbia University Medical Center, Columbia University, New York, New York. Herbert Irving Comprehensive Cancer Center, Columbia University Medical Center, Columbia University, New York, New York. Department of Surgery, Columbia University Medical Center, Columbia University, New York, New York.
Carrie J
CJ
Shawber
Department of Obstetrics/Gynecology, Columbia University Medical Center, Columbia University, New York, New York. Department of Pathology and Cellular Biology, Columbia University Medical Center, Columbia University, New York, New York. Herbert Irving Comprehensive Cancer Center, Columbia University Medical Center, Columbia University, New York, New York. jkk9@columbia.edu.
Jan
J
Kitajewski
D020533:Angiogenesis Inhibitors / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D000818:Animals; D005260:Female; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009369:Neoplasms / Q000098:blood supply* / Q000378:metabolism / Q000628:therapy*; D009389:Neovascularization, Pathologic / Q000378:metabolism / Q000628:therapy; D011485:Protein Binding; D051881:Receptor, Notch1 / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D051880:Receptors, Notch / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D015398:Signal Transduction
D020533:Angiogenesis Inhibitors; D007141:Immunoglobulin Fc Fragments; D036341:Intercellular Signaling Peptides and Proteins; D051881:Receptor, Notch1; D051880:Receptors, Notch; D011993:Recombinant Fusion Proteins