Hypoxia (low oxygen) and Notch signaling are 2 important regulators of vascular development, but how they interact in controlling the choice between arterial and venous fates for endothelial cells during vasculogenesis is less well understood. In this report, we show that hypoxia and Notch signaling intersect in promotion of arterial differentiation. Hypoxia upregulated expression of the Notch ligand Dll4 and increased Notch signaling in a process requiring the vasoactive hormone adrenomedullin. Notch signaling also upregulated Dll4 expression, leading to a positive feedback loop sustaining Dll4 expression and Notch signaling. In addition, hypoxia-mediated upregulation of the arterial marker genes Depp, connexin40 (Gja5), Cxcr4, and Hey1 required Notch signaling. In conclusion, the data reveal an intricate interaction between hypoxia and Notch signaling in the control of endothelial cell differentiation, including a hypoxia/adrenomedullin/Dll4 axis that initiates Notch signaling and a requirement for Notch signaling to effectuate hypoxia-mediated induction of the arterial differentiation program.
Get full access to this article
View all access options for this article.
References
1.
RisauW, FlammeI. 1995. Vasculogenesis. Ann Rev Cell Dev Biol, 11:73–91.
2.
CarmelietP. 2003. Angiogenesis in health and disease. Nat Med, 9:653–660.
3.
AtkinsGB, JainMK, HamikA. 2011. Endothelial differentiation: molecular mechanisms of specification and heterogeneity. Arterioscler Thromb Vasc Biol, 31:1476–1484.
LannerF, SohlM, FarneboF. 2007. Functional arterial and venous fate is determined by graded VEGF signaling and notch status during embryonic stem cell differentiation. Arterioscler Thromb Vasc Biol, 27:487–493.
6.
PetersonRT, ShawSY, PetersonTA, MilanDJ, ZhongTP, SchreiberSL, MacRaeCA, FishmanMC. 2004. Chemical suppression of a genetic mutation in a zebrafish model of aortic coarctation. Nat Biotechnol, 22:595–599.
7.
SeoS, FujitaH, NakanoA, KangM, DuarteA, KumeT. 2006. The forkhead transcription factors, Foxc1 and Foxc2, are required for arterial specification and lymphatic sprouting during vascular development. Dev Biol, 294:458–470.
8.
WeinsteinBM, StempleDL, DrieverW, FishmanMC. 1995. Gridlock, a localized heritable vascular patterning defect in the zebrafish. Nat Med, 1:1143–1147.
9.
CoultasL, ChawengsaksophakK, RossantJ. 2005. Endothelial cells and VEGF in vascular development. Nature, 438:937–945.
10.
LendahlU, LeeKL, YangH, PoellingerL. 2009. Generating specificity and diversity in the transcriptional response to hypoxia. Nat Rev Genet, 10:821–832.
GaleNW, DominguezMG, NogueraI, PanL, HughesV, ValenzuelaDM, MurphyAJ, AdamsNC, LinHCet al.2004. Haploinsufficiency of delta-like 4 ligand results in embryonic lethality due to major defects in arterial and vascular development. Proc Natl Acad Sci U S A, 101:15949–15954.
17.
XueY, GaoX, LindsellCE, NortonCR, ChangB, HicksC, Gendron-MaguireM, RandEB, WeinmasterG, GridleyT. 1999. Embryonic lethality and vascular defects in mice lacking the Notch ligand Jagged1. Hum Mol Genet, 8:723–730.
18.
OkaC, NakanoT, WakehamA, de la PompaJL, MoriC, SakaiT, OkazakiS, KawaichiM, ShiotaK, MakTW, HonjoT. 1995. Disruption of the mouse RBP-J kappa gene results in early embryonic death. Development, 121:3291–3301.
19.
GustafssonMV, ZhengX, PereiraT, GradinK, JinS, LundkvistJ, RuasJL, PoellingerL, LendahlU, BondessonM. 2005. Hypoxia requires notch signaling to maintain the undifferentiated cell state. Dev Cell, 9:617–628.
20.
SahlgrenC, GustafssonMV, JinS, PoellingerL, LendahlU. 2008. Notch signaling mediates hypoxia-induced tumor cell migration and invasion. Proc Natl Acad Sci U S A, 105:6392–6397.
21.
ColemanML, McDonoughMA, HewitsonKS, ColesC, MecinovicJ, EdelmannM, CookKM, CockmanME, LancasterDEet al.2007. Asparaginyl hydroxylation of the Notch ankyrin repeat domain by factor inhibiting hypoxia-inducible factor. J Biol Chem, 282:24027–24038.
22.
DiezH, FischerA, WinklerA, HuCJ, HatzopoulosAK, BreierG, GesslerM. 2007. Hypoxia-mediated activation of Dll4-Notch-Hey2 signaling in endothelial progenitor cells and adoption of arterial cell fate. Exp Cell Res, 313:1–9.
23.
DongX, WangYS, DouGR, HouHY, ShiYY, ZhangR, MaK, WuL, YaoLB, CaiY, ZhangJ. 2011. Influence of Dll4 via HIF-1alpha-VEGF signaling on the angiogenesis of choroidal neovascularization under hypoxic conditions. PLoS One, 6:e18481.
24.
CarmelietP, FerreiraV, BreierG, PollefeytS, KieckensL, GertsensteinM, FahrigM, VandenhoeckA, HarpalKet al.1996. Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature, 380:435–439.
25.
DasD, LannerF, MainH, AnderssonER, BergmannO, SahlgrenC, HeldringN, HermansonO, HanssonEM, LendahlU. 2010. Notch induces cyclin-D1-dependent proliferation during a specific temporal window of neural differentiation in ES cells. Dev Biol, 348:153–166.
26.
MainH, LeeKL, YangH, Haapa-PaananenS, EdgrenH, JinS, SahlgrenC, KallioniemiO, PoellingerL, LimB, LendahlU. 2010. Interactions between Notch- and hypoxia-induced transcriptomes in embryonic stem cells. Exp Cell Res, 316:1610–1624.
27.
PajusolaK, KunnapuuJ, VuorikoskiS, SoronenJ, AndreH, PereiraT, KorpisaloP, Yla-HerttualaS, PoellingerL, AlitaloK. 2005. Stabilized HIF-1 alpha is superior to VEGF for angiogenesis in skeletal muscle via adeno-associated virus gene transfer. FASEB J, 19:1365.
28.
JakobssonL, KreugerJ, HolmbornK, LundinL, ErikssonI, KjellenL, Claesson-WelshL. 2006. Heparan sulfate in trans potentiates VEGFR-mediated angiogenesis. Dev Cell, 10:625–634.
29.
ChuaSW, VijayakumarP, NissomPM, YamCY, WongVV, YangH. 2006. A novel normalization method for effective removal of systematic variation in microarray data. Nucleic Acids Res, 34:e38.
30.
LiX, EdholmD, LannerF, BreierG, FarneboF, DimbergA, Claesson-WelshL. 2007. Lentiviral rescue of vascular endothelial growth factor receptor-2 expression in flk1−/− embryonic stem cells shows early priming of endothelial precursors. Stem Cells, 25:2987–2995.
31.
YehHI, DupontE, CoppenS, RotheryS, SeversNJ. 1997. Gap junction localization and connexin expression in cytochemically identified endothelial cells of arterial tissue. J Histochem Cytochem, 45:539–550.
32.
ShinD, AndersonDJ. 2005. Isolation of arterial-specific genes by subtractive hybridization reveals molecular heterogeneity among arterial endothelial cells. Dev Dyn, 233:1589–1604.
33.
KrebsLT, XueY, NortonCR, ShutterJR, MaguireM, SundbergJP, GallahanD, ClossonV, KitajewskiJet al.2000. Notch signaling is essential for vascular morphogenesis in mice. Genes Dev, 14:1343–1352.
34.
WuQY, DrouetL, CarrierJL, RothschildC, BerardM, RouaultC, CaenJP, MeyerD. 1987. Differential distribution of von Willebrand factor in endothelial cells. Comparison between normal pigs and pigs with von Willebrand disease. Arteriosclerosis, 7:47–54.
35.
Mecha DisassaN, Styp-RekowskaB, HinzB, Da Silva-AzevedoL, PriesAR, ZakrzewiczA. 2009. Differential expression of VEGFA, TIE2, and ANG2 but not ADAMTS1 in rat mesenteric microvascular arteries and veins. Physiol Res, 58:193–202.
36.
Yurugi-KobayashiT, ItohH, SchroederT, NakanoA, NarazakiG, KitaF, YanagiK, Hiraoka-KanieM, InoueEet al.2006. Adrenomedullin/cyclic AMP pathway induces Notch activation and differentiation of arterial endothelial cells from vascular progenitors. Arterioscler Thromb Vasc Biol, 26:1977–1984.
37.
KalogerisTJ, KevilCG, LarouxFS, CoeLL, PhiferTJ, AlexanderJS. 1999. Differential monocyte adhesion and adhesion molecule expression in venous and arterial endothelial cells. Am J Physiol, 276:L9–L19.
JiaW, LiH, HeYW. 2005. The extracellular matrix protein mindin serves as an integrin ligand and is critical for inflammatory cell recruitment. Blood, 106:3854–3859.
41.
KatoH, TaniguchiY, KurookaH, MinoguchiS, SakaiT, Nomura-OkazakiS, TamuraK, HonjoT. 1997. Involvement of RBP-J in biological functions of mouse Notch1 and its derivatives. Development, 124:4133–4141.
42.
YouLR, LinFJ, LeeCT, DeMayoFJ, TsaiMJ, TsaiSY. 2005. Suppression of Notch signalling by the COUP-TFII transcription factor regulates vein identity. Nature, 435:98–104.
43.
CaronKM, SmithiesO. 2001. Extreme hydrops fetalis and cardiovascular abnormalities in mice lacking a functional Adrenomedullin gene. Proc Natl Acad Sci U S A, 98:615–619.
44.
DackorRT, Fritz-SixK, DunworthWP, GibbonsCL, SmithiesO, CaronKM. 2006. Hydrops fetalis, cardiovascular defects, and embryonic lethality in mice lacking the calcitonin receptor-like receptor gene. Mol Cell Biol, 26:2511–2518.
45.
NicoliS, TobiaC, GualandiL, De SenaG, PrestaM. 2008. Calcitonin receptor-like receptor guides arterial differentiation in zebrafish. Blood, 111:4965–4972.
46.
GuidolinD, AlbertinG, SpinazziR, SoratoE, MascarinA, CavalloD, AntonelloM, RibattiD. 2008. Adrenomedullin stimulates angiogenic response in cultured human vascular endothelial cells: involvement of the vascular endothelial growth factor receptor 2. Peptides, 29:2013–2023.
47.
NakayamaM, TakahashiK, MurakamiO, ShiratoK, ShibaharaS. 1998. Induction of adrenomedullin by hypoxia and cobalt chloride in human colorectal carcinoma cells. Biochem Biophys Res Commun, 243:514–517.
48.
Cormier-RegardS, NguyenSV, ClaycombWC. 1998. Adrenomedullin gene expression is developmentally regulated and induced by hypoxia in rat ventricular cardiac myocytes. J Biol Chem, 273:17787–17792.
49.
LiJL, SainsonRC, OonCE, TurleyH, LeekR, SheldonH, BridgesE, ShiW, SnellCet al.2011. DLL4-Notch signaling mediates tumor resistance to anti-VEGF therapy in vivo. Cancer Res, 71:6073–6083.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
