HMG-1/Y Regulation of NOS2 Expression in Endotoxemia

HMG-1/Y 在内毒素血症中 NOS2 表达的调节

• 批准号: 6886305
• 负责人: Rebecca M Baron
• 金额: $ 12.85万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6886305
• 关键词: enzyme biosynthesis; genetic regulatory element; genetically modified animals; immunoprecipitation; interferon gamma; laboratory mouse; lipopolysaccharides; macrophage; nitric oxide; nitric oxide synthase; protein binding; septicemia; tissue /cell culture; transcription factor; transfection; vascular smooth muscle

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：败血症是一种疾病过程，代表严重感染的全身反应，导致高发病率和死亡率。在一部分脓毒症患者中，细菌释放毒素如脂多糖（LPS）并激活炎症反应，包括释放干扰素(IFN)- γ等细胞因子。一氧化氮合酶（NOS2）的诱导形式通过过量产生一氧化氮（NO）在内毒素血症中起重要作用，这与内毒素血症的许多病理生理机制有关。该提案的目标是：(a)获得研究内毒素血症条件下建筑转录因子高迁移率组- i /Y （HMG-I/Y）在NOS2表达中的作用的技能和知识；(b)获得教导他人的经验；(c)获得领导独立研究小组所需的专业知识。在布里格姆妇女医院进行的培训将包括参加几项有组织的研究活动，从而实现科学独立。该实验室配备了细胞和分子生物学、动物生理学和组织学研究所需的资源。我们提出的工作的总体假设是HMG-I/Y促进LPS/ ifn - γ协同诱导NOS2，并且可能是内毒素血症中NO产生的重要调节剂。在AIM 1中，我们将通过体外实验确定HMG-I/Y是否与LPS/ ifn - γ诱导NOS2的重要转录因子相互作用。在AIM 2中，我们将确定在体外和体内改变HMG-I/Y与DNA的结合是否会改变内毒素血症条件下NOS2基因的表达和动物的存活。在AIM 3中，我们将通过在血管中表达显性阴性HMG-I/Y的转基因小鼠，阐明HMG-I/Y的作用及其对内毒素血症中NOS2表达和动物存活的影响。这些实验将使我们能够确定HMG-I/Y在内毒素血症期间调节NOS2表达的作用，并可能揭示改善败血症预后的新靶点。