Adrenomedullin/cyclic AMP pathway induces Notch activation and differentiation of arterial endothelial cells from vascular progenitors.
OBJECTIVE
The acquisition of arterial or venous identity is highlighted in vascular development. Previously, we have reported an embryonic stem (ES) cell differentiation system that exhibits early vascular development using vascular endothelial growth factor (VEGF) receptor-2 (VEGFR2)-positive cells as common vascular progenitors. In this study, we constructively induced differentiation of arterial and venous endothelial cells (ECs) in vitro to elucidate molecular mechanisms of arterial-venous specification.


RESULTS
ECs were induced from VEGFR2+ progenitor cells with various conditions. VEGF was essential to induce ECs. Addition of 8bromo-cAMP or adrenomedullin (AM), an endogenous ligand-elevating cAMP, enhanced VEGF-induced EC differentiation. Whereas VEGF alone mainly induced venous ECs, 8bromo-cAMP (or AM) with VEGF supported substantial induction of arterial ECs. Stimulation of cAMP pathway induced Notch signal activation in ECs. The arterializing effect of VEGF and cAMP was abolished in recombination recognition sequence binding protein at the Jkappa site deficient ES cells lacking Notch signal activation or in ES cells treated with gamma-secretase inhibitor. Nevertheless, forced Notch activation by the constitutively active Notch1 alone did not induce arterial ECs.


CONCLUSIONS
Adrenomedullin/cAMP is a novel signaling pathway to activate Notch signaling in differentiating ECs. Coordinated signaling of VEGF, Notch, and cAMP is required to induce arterial ECs from vascular progenitors.
Yurugi-Kobayashi
Takami
T
Laboratory of Stem Cell Differentiation, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507 Japan.
Itoh
Hiroshi
H
Schroeder
Timm
T
Nakano
Akiko
A
Narazaki
Genta
G
Kita
Fumiyo
F
Yanagi
Kentoku
K
Hiraoka-Kanie
Mina
M
Inoue
Emi
E
Ara
Toshiaki
T
Nagasawa
Takashi
T
Just
Ursula
U
Nakao
Kazuwa
K
Nishikawa
Shin-ichi
S
Yamashita
Jun K
JK
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D053607:Adrenomedullin; D000818:Animals; D001158:Arteries / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism*; D004357:Drug Synergism; D042783:Endothelial Cells / Q000166:cytology*; D051379:Mice; D010455:Peptides / Q000378:metabolism* / Q000494:pharmacology; D051880:Receptors, Notch / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D013234:Stem Cells / Q000166:cytology*; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D014680:Veins / Q000166:cytology
D010455:Peptides; D051880:Receptors, Notch; D042461:Vascular Endothelial Growth Factor A; D053607:Adrenomedullin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000242:Cyclic AMP