Sepsis is a disease process representing the systemic response to severe infection and causes high morbidity and mortality. In a subset of sepsis patients, toxins such as lipopolysaccharide (LPS) are released by bacteria and activate an inflammatory response, including the release of cytokines such as interferon (IFN)-gamma. The inducible form of nitric oxide synthase (NOS2) plays an important role in endotoxemia through overproduction of nitric oxide (NO), which has been implicated in a number of pathophysiologic mechanisms of endotoxemia. The objectives of the proposal are: (a) to gain skills and knowledge in the investigation of the role of the architectural transcription factor high mobility group-I/Y (HMG-I/Y) in NOS2 expression under conditions of endotoxemia; (b) to gain experience in teaching others; and (c) to achieve the necessary expertise to lead an independent research group. This training at Brigham and Women's Hospital will incorporate the participation in several structured research activities leading to scientific independence. The laboratory is equipped with the resources required for research in cellular and molecular biology, animal physiology, and histology. The overall hypothesis of our proposed work is that HMG-I/Y facilitates the LPS/IFN-gamma synergistic induction of NOS2 and may be an important modulator of NO production in endotoxemia.
In AIM 1 we will determine whether HMG-I/Y interacts with the transcription factors important for LPS/IFN-gamma induction of NOS2 using in vitro assays.
In AIM 2, we will determine whether altering HMG-I/Y binding to DNA in vitro and in vivo alters NOS2 gene expression and animal survival under conditions of endotoxemia.
In AIM 3, we will elucidate the role of HMG-I/Y and its effect on NOS2 expression and animal survival during endotoxemia using a transgenic mouse expressing a dominant-negative form of HMG-I/Y in the vasculature. These experiments will allow us to determine the role of HMG-I/Y in regulating NOS2 expression during endotoxemia and may reveal a novel target for improving outcomes from sepsis. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08AI054465-03
Application #
6886305
Study Section
Allergy & Clinical Immunology-1 (AITC)
Program Officer
Prograis, Lawrence J
Project Start
2003-05-01
Project End
2008-04-30
Budget Start
2005-05-01
Budget End
2006-04-30
Support Year
3
Fiscal Year
2005
Total Cost
$128,520
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
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Baron, Rebecca M; Lopez-Guzman, Silvia; Riascos, Dario F et al. (2010) Distamycin A inhibits HMGA1-binding to the P-selectin promoter and attenuates lung and liver inflammation during murine endotoxemia. PLoS One 5:e10656
Grant, Marianne A; Baron, Rebecca M; Macias, Alvaro A et al. (2009) Netropsin improves survival from endotoxaemia by disrupting HMGA1 binding to the NOS2 promoter. Biochem J 418:103-12
Takamiya, Rina; Baron, Rebecca M; Yet, Shaw-Fang et al. (2008) High mobility group A1 protein mediates human nitric oxide synthase 2 gene expression. FEBS Lett 582:810-4
Baron, Beverly W; Zeleznik-Le, Nancy; Baron, Miriam J et al. (2007) Repression of the PDCD2 gene by BCL6 and the implications for the pathogenesis of human B and T cell lymphomas. Proc Natl Acad Sci U S A 104:7449-54
Wang, Ying; Baron, Rebecca M; Zhu, Guangli et al. (2006) PU.1 regulates cathepsin S expression in professional APCs. J Immunol 176:275-83
Fredenburgh, L E; Baron, R M; Carvajal, I M et al. (2005) Absence of heme oxygenase-1 expression in the lung parenchyma exacerbates endotoxin-induced acute lung injury and decreases surfactant protein-B levels. Cell Mol Biol (Noisy-le-grand) 51:513-20
Baron, Rebecca M; Carvajal, Irvith M; Fredenburgh, Laura E et al. (2004) Nitric oxide synthase-2 down-regulates surfactant protein-B expression and enhances endotoxin-induced lung injury in mice. FASEB J 18:1276-8
Baron, Rebecca M; Carvajal, Irvith M; Liu, Xiaoli et al. (2004) Reduction of nitric oxide synthase 2 expression by distamycin A improves survival from endotoxemia. J Immunol 173:4147-53