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: 7227530
• 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 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7227530
• 关键词: AT Rich Sequence; Address; All Sites; Animals; Bacteria; Binding; Binding Sites; Blood Pressure; Blood Vessels; Carboxypeptidase; Clinical; Co-Immunoprecipitations; Complex; Condition; Consensus; Cultured Cells; DNA; DNA Binding; DNA Sequence; DNA-Binding Proteins; Development; Disease; Disruption; Distamycin; Dominant-Negative Mutation; Educational process of instructing; Electrophoretic Mobility Shift Assay; End Point; Endotoxemia; Gene Expression; Gene Proteins; Histology; Hospitals; IRF1 gene; In Vitro; Infection; Inflammatory Response; Interferon Type II; Interferons; Interleukins; Investigation; Knowledge; Laboratories; Lead; Lipopolysaccharides; Maintenance; Measurement; Molecular and Cellular Biology; Morbidity - disease rate; Mus; NOS2A gene; Nitric Oxide; Nitric Oxide Synthase; Nuclear; Nuclear Protein; Nuclear Proteins; Numbers; Organ; Outcome; Patients; Physiology; Play; Process; Production; Promoter Regions; Protein Overexpression; Regulation; Research; Research Activity; Research Personnel; Resources; Role; Sepsis; Site; Smooth Muscle Myocytes; Structure; Testing; Tissues; Toxin; Training; Trans-Activators; Transfection; Transgenic Mice; Up-Regulation; Woman; Work; cis acting element; cytokine; experience; gene induction; human NOS2A protein; immunoregulation; improved; in vitro Assay; in vivo; inhibitor/antagonist; macrophage; mortality; novel; programs; promoter; research study; response; skills; transcription factor; transgene expression

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（由申请方提供）：脓毒症是一种疾病过程，代表对严重感染的全身反应，并导致高发病率和死亡率。在败血症患者的子集中，毒素如脂多糖（LPS）由细菌释放并激活炎症反应，包括释放细胞因子如干扰素（IFN）-γ。诱导型一氧化氮合酶（inducible form of nitric oxide synthase，NOS 2）通过过量产生一氧化氮（nitric oxide，NO）在内毒素血症中起重要作用，参与了内毒素血症的多种病理生理机制。该提案的目的是：（a）获得技能和知识，在调查的作用，建筑转录因子高迁移率组-I/Y（HMG-I/Y）在NOS 2表达条件下的内毒素血症;（B）获得教学经验;和（c）实现必要的专业知识，领导一个独立的研究小组。在布里格姆妇女医院的培训将包括参与几项结构化的研究活动，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NOS 2，并可能是内毒素血症中NO产生的重要调节剂。在AIM 1中，我们将使用体外测定来确定HMG-I/Y是否与对于LPS/IFN-γ诱导NOS 2重要的转录因子相互作用。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NOS 2基因表达和内毒素血症条件下动物的存活率。在AIM 3中，我们将阐明HMG-I/Y的作用及其对NOS 2表达和内毒素血症期间动物存活的影响，使用在血管系统中表达显性阴性形式HMG-I/Y的转基因小鼠。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NOS 2表达的作用，并可能揭示改善脓毒症结局的新靶点。