The candidate has a three-year post-doctoral experience and is currently being proposed for an Associate Research Scientist appointment at the School of Medicine of Yale University. This application for a mentored K01 award is submitted with the goal of providing the candidate with the further training experience to function as an independent investigator working in the field of molecular biology and biochemistry of the liver. The project's long-term objective is to advance our understanding of nitric oxide (NO) signaling in liver physiology and pathophysiology. This is important for developing novel strategies to treat complications associated with liver diseases, since abnormal NO production by endothelial NO synthase (eNOS) plays a key role in vascular abnormalities observed in portal hypertension in liver cirrhosis. S-nitrosylation of proteins is an emerging area of investigation for NO-mediated physiological responses. We propose to investigate the mechanisms of S-nitrosylation, focusing on subcellular localization of eNOS and NO production, and identify the profiles of S-nitrosylated proteins in endothelial cells and liver cells using a proteomic approach. Our preliminary results suggest that there is a high concentration of NO gas at the eNOS localized area on the Golgi. This leads us to hypothesize that a gradient of NO formed in proximity to eNOS on the Golgi may be favorable to S-nitrosylation of proteins. The candidate proposes to 1) Elucidate if eNOS generates a gradient of NO in living cells, 2) Identify S-nitrosylated proteins in/on the Golgi complex using a proteomic approach, and 3) Examine protein S-nitrosylation in sinusoidal endothelial cells and hepatocytes. The Section of Digestive Diseases and the Department of Pharmacology as well as the Department of Cell Biology at Yale University are ideal for carrying out these studies because of the quality of their faculty, their experience as mentors, and the core facility at the Yale Liver Center. The School of Medicine has pledged protected time for the candidate during this further training period prior to functioning as an independent investigator.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01DK067933-05
Application #
7431778
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Program Officer
Podskalny, Judith M,
Project Start
2004-08-15
Project End
2010-06-30
Budget Start
2008-07-01
Budget End
2010-06-30
Support Year
5
Fiscal Year
2008
Total Cost
$136,755
Indirect Cost
Name
Yale University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Huang, Hui-Chun; Haq, Omar; Utsumi, Teruo et al. (2012) Intestinal and plasma VEGF levels in cirrhosis: the role of portal pressure. J Cell Mol Med 16:1125-33
Sangwung, Panjamaporn; Greco, Todd M; Wang, Yanzhuang et al. (2012) Proteomic identification of S-nitrosylated Golgi proteins: new insights into endothelial cell regulation by eNOS-derived NO. PLoS One 7:e31564
Iwakiri, Yasuko (2012) Endothelial dysfunction in the regulation of cirrhosis and portal hypertension. Liver Int 32:199-213
Zhang, Dahai; Utsumi, Teruo; Huang, Hui-Chun et al. (2011) Reticulon 4B (Nogo-B) is a novel regulator of hepatic fibrosis. Hepatology 53:1306-15
Iwakiri, Yasuko (2011) S-nitrosylation of proteins: a new insight into endothelial cell function regulated by eNOS-derived NO. Nitric Oxide 25:95-101
Iwakiri, Yasuko (2007) The molecules: mechanisms of arterial vasodilatation observed in the splanchnic and systemic circulation in portal hypertension. J Clin Gastroenterol 41:S288-94
Loureiro-Silva, Mauricio R; Iwakiri, Yasuko; Abraldes, Juan G et al. (2006) Increased phosphodiesterase-5 expression is involved in the decreased vasodilator response to nitric oxide in cirrhotic rat livers. J Hepatol 44:886-93
Iwakiri, Yasuko; Groszmann, Roberto J (2006) The hyperdynamic circulation of chronic liver diseases: from the patient to the molecule. Hepatology 43:S121-31
Abraldes, Juan G; Iwakiri, Yasuko; Loureiro-Silva, Mauricio et al. (2006) Mild increases in portal pressure upregulate vascular endothelial growth factor and endothelial nitric oxide synthase in the intestinal microcirculatory bed, leading to a hyperdynamic state. Am J Physiol Gastrointest Liver Physiol 290:G980-7
Iwakiri, Yasuko; Satoh, Ayano; Chatterjee, Suvro et al. (2006) Nitric oxide synthase generates nitric oxide locally to regulate compartmentalized protein S-nitrosylation and protein trafficking. Proc Natl Acad Sci U S A 103:19777-82