(Verbatim from the application): The long-term goals are to identify at the biochemical and cellular levels the signal transduction pathway(s) which link Angiotensin II (Ang II) to the production of nitric oxide (NO), a potent vasodilator, in endothelial cells. This effect of Ang II is in sharp contrast to its well-known role in the cardiovascular system as a potent vasoconstrictor. The underlying hypothesis is that these apparently contradictory effects of Ang II reflect signaling events coupled to the AT1 and AT2 receptors, respectively, and that the vasodilatory effects of Ang II are mediated by the AT2 receptors via currently unidentified signaling pathways. This emerges from the novel findings that Ang II stimulates NO production in pulmonary endothelial cells, via increased expression of endothelial nitric oxide synthase (eNOS). Preliminary data indicate that this is mediated via the AT2 receptor, and support a mechanism by which Ang II binding to a G protein-coupled AT2 receptor leads to an increase in tyrosine phosphorylation, which ultimately results in increased eNOS protein expression. Furthermore, signaling via the AT1 receptor appears to negatively regulate eNOS protein expression.
Aim I is directed at the characterization of AT1/AT2 receptor subtype distribution on endothelial cells, and further investigation of the mechanism of Ang II-stimulated eNOS mRNA and protein expression. The role of the Ang II receptor subtypes in mediating Ang II dependent enhancement of hypoxic vasoconstriction in pulmonary endothelium will be determined. The goal of the second aim is to identify the pertussis toxin-sensitive G protein that is linked to AT2 receptor.
In Aim 3 the tyrosine kinase(s) that are activated, or protein tyrosine phosphatase(s) that are inhibited, and that provide the signaling linkage between Ang II and NO production, will be identified. Receptor antagonists, binding studies, and analysis of mRNA and protein levels will be used to identify the receptor subtypes. Pharmacological inhibitors, antisense oligonucleotides and antibody immunoneutralization experiments will be used to investigate the role of various signaling components (Mg II receptors, GTP binding proteins and protein kinases). These studies could provide a molecular basis for a novel hypothesis that would rationalize the physiologically important but opposing effects of Ang II on smooth muscle cells versus endothelial cells. These studies are of significant medical importance to heart and pulmonary disease, which involve pathological changes in blood pressure.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL063182-03
Application #
6627535
Study Section
Experimental Cardiovascular Sciences Study Section (ECS)
Program Officer
Gail, Dorothy
Project Start
2001-01-01
Project End
2004-12-31
Budget Start
2003-01-01
Budget End
2003-12-31
Support Year
3
Fiscal Year
2003
Total Cost
$313,000
Indirect Cost
Name
New York Medical College
Department
Biochemistry
Type
Schools of Medicine
DUNS #
041907486
City
Valhalla
State
NY
Country
United States
Zip Code
10595
Li, Jianyu; Zhao, Xiangmin; Li, Xinmei et al. (2007) Angiotensin II type 2 receptor-dependent increases in nitric oxide synthase expression in the pulmonary endothelium is mediated via a G alpha i3/Ras/Raf/MAPK pathway. Am J Physiol Cell Physiol 292:C2185-96
Oeckler, Richard A; Arcuino, Elizabeth; Ahmad, Mansoor et al. (2005) Cytosolic NADH redox and thiol oxidation regulate pulmonary arterial force through ERK MAP kinase. Am J Physiol Lung Cell Mol Physiol 288:L1017-25
Zhao, Xiangmin; Li, Xinmei; Trusa, Sandra et al. (2005) Angiotensin type 1 receptor is linked to inhibition of nitric oxide production in pulmonary endothelial cells. Regul Pept 132:113-22
Olson, Susan; Oeckler, Richard; Li, Xinmei et al. (2004) Angiotensin II stimulates nitric oxide production in pulmonary artery endothelium via the type 2 receptor. Am J Physiol Lung Cell Mol Physiol 287:L559-68
Li, Xinmei; Lerea, Kenneth M; Li, Jianyu et al. (2004) Src kinase mediates angiotensin II-dependent increase in pulmonary endothelial nitric oxide synthase. Am J Respir Cell Mol Biol 31:365-72