The differentiation of arterial vs. venous endothelial cells (EC) occurs early during vertebrate cardiovascular development, and appears to involve interplay of both intrinsic genetic factors and environmental stimuli. Biomechanical forces generated by blood flow are strong modulators of the endothelial phenotype in health and disease. Our preliminary studies in vitro have revealed that venous EC can be modulated towards an arterial identity when exposed to a biomechanical stimulus that simulates components of the arterial circulation. In addition, a mouse model of venous arterialization has demonstrated the expression of arterial identity markers in EC of veins exposed to an arterial circulation. Thus, we hypothesize that hemodynamic forces can act as cell differentiation stimuli controlling the establishment and maintenance of endothelial arterial and venous identities. To test this hypothesis we propose three Specific Aims.
In Aim 1, the transcriptional programs evoked when cultured human venous EC are exposed to arterial shear stress waveforms will be characterized and compared with those observed in EC freshly isolated from human arteries and veins.
In Aim 2, the role of Notch signaling in flow-mediated venous-to-arterial endothelial differentiation will be investigated utilizing gain-of-function and loss-of-function mutants of Notch and its downstream effectors.
In Aim 3, a novel mouse model will be used to test the in vivo relevance of Notch and two of its transcriptional targets, the bHLH transcription factors Hey-1 and Hey-2, in venous arterialization. Our long-term goal is to understand the flow-mediated molecular and cellular mechanisms that control arterial vs. venous specification in the vascular system. These studies should provide important mechanistic insights into the control of endothelial identity by hemodynamic forces, and may suggest new therapeutic strategies for diseases in which dysregulation of this process leads to serious pathologies, including saphenous vein bypass graft failure and arterio-venous malformations.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL076686-04
Application #
7193403
Study Section
Experimental Cardiovascular Sciences Study Section (ECS)
Program Officer
Goldman, Stephen
Project Start
2004-04-01
Project End
2009-03-31
Budget Start
2007-04-01
Budget End
2008-03-31
Support Year
4
Fiscal Year
2007
Total Cost
$282,840
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Gimbrone Jr, Michael A; García-Cardeña, Guillermo (2013) Vascular endothelium, hemodynamics, and the pathobiology of atherosclerosis. Cardiovasc Pathol 22:9-15
Wu, Wei; Xiao, Han; Laguna-Fernandez, Andrés et al. (2011) Flow-Dependent Regulation of Kruppel-Like Factor 2 Is Mediated by MicroRNA-92a. Circulation 124:633-41
Adamo, Luigi; Garcia-Cardena, Guillermo (2011) Directed stem cell differentiation by fluid mechanical forces. Antioxid Redox Signal 15:1463-73
Gracia-Sancho, Jorge; Russo, Lucia; Garcia-Caldero, Hector et al. (2011) Endothelial expression of transcription factor Kruppel-like factor 2 and its vasoprotective target genes in the normal and cirrhotic rat liver. Gut 60:517-24
Bu, De-xiu; Tarrio, Margarite; Grabie, Nir et al. (2010) Statin-induced Krüppel-like factor 2 expression in human and mouse T cells reduces inflammatory and pathogenic responses. J Clin Invest 120:1961-70
Gracia-Sancho, Jorge; Villarreal Jr, Guadalupe; Zhang, Yuzhi et al. (2010) Flow cessation triggers endothelial dysfunction during organ cold storage conditions: strategies for pharmacologic intervention. Transplantation 90:142-9
Villarreal Jr, Guadalupe; Zhang, Yuzhi; Larman, H Benjamin et al. (2010) Defining the regulation of KLF4 expression and its downstream transcriptional targets in vascular endothelial cells. Biochem Biophys Res Commun 391:984-9
Gracia-Sancho, Jorge; Villarreal Jr, Guadalupe; Zhang, Yuzhi et al. (2010) Activation of SIRT1 by resveratrol induces KLF2 expression conferring an endothelial vasoprotective phenotype. Cardiovasc Res 85:514-9
Young, Angela; Wu, Wei; Sun, Wei et al. (2009) Flow activation of AMP-activated protein kinase in vascular endothelium leads to Krüppel-like factor 2 expression. Arterioscler Thromb Vasc Biol 29:1902-8
Liu, Meng; Kluger, Martin S; D'Alessio, Alessio et al. (2008) Regulation of arterial-venous differences in tumor necrosis factor responsiveness of endothelial cells by anatomic context. Am J Pathol 172:1088-99

Showing the most recent 10 out of 14 publications