Notch alters VEGF responsiveness in human and murine endothelial cells by direct regulation of VEGFR-3 expression.
The Notch family of cell surface receptors and its ligands are highly conserved proteins that regulate cell fate determination, including those involved in mammalian vascular development. We report that Notch induces VEGFR-3 expression in vitro in human endothelial cells and in vivo in mice. In vitro, Notch in complex with the DNA-binding protein CBF-1/suppressor of hairless/Lag1 (CSL) bound the VEGFR-3 promoter and transactivated VEGFR-3 specifically in endothelial cells. Through induction of VEGFR-3, Notch increased endothelial cell responsiveness to VEGF-C, promoting endothelial cell survival and morphological changes. In vivo, VEGFR-3 was upregulated in endothelial cells with active Notch signaling. Mice heterozygous for null alleles of both Notch1 and VEGFR-3 had significantly reduced viability and displayed midgestational vascular patterning defects analogous to Notch1 nullizygous embryos. We found that Notch1 and Notch4 were expressed in normal and tumor lymphatic endothelial cells and that Notch1 was activated in lymphatic endothelium of invasive mammary micropapillary carcinomas. These results demonstrate that Notch1 and VEGFR-3 interact genetically, that Notch directly induces VEGFR-3 in blood endothelial cells to regulate vascular development, and that Notch may function in tumor lymphangiogenesis.
Shawber
Carrie J
CJ
Department of Obstetrics and Gynecology, Columbia University Medical Center, New York, New York 10032, USA.
Funahashi
Yasuhiro
Y
Francisco
Esther
E
Vorontchikhina
Marina
M
Kitamura
Yukari
Y
Stowell
Stephanie A
SA
Borisenko
Valeriya
V
Feirt
Nikki
N
Podgrabinska
Simona
S
Shiraishi
Kazuko
K
Chawengsaksophak
Kallayanee
K
Rossant
Janet
J
Accili
Domenico
D
Skobe
Mihaela
M
Kitajewski
Jan
J
D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D048430:Cell Shape; D002470:Cell Survival; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000033:anatomy & histology / Q000502:physiology; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000235:genetics / Q000378:metabolism*
D051880:Receptors, Notch; D042461:Vascular Endothelial Growth Factor A; D040321:Vascular Endothelial Growth Factor Receptor-3