In Vivo Role of NO in Mucosal Inflammation and Cancer

NO 在粘膜炎症和癌症中的体内作用

基本信息

批准号：
6990335
负责人：
DAVID B SCHAUER
金额：
$ 13.17万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-13 至 2008-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6990335
关键词：
Helicobacter T cell receptor carcinogenesis cytokine cytotoxicity enzyme induction /repression enzyme inhibitors genetically modified animals histopathology inflammatory bowel diseases laboratory mouse macrophage neutrophil nitric oxide nitric oxide synthase nitrogen metabolism oxidative stress

项目摘要

This Project will characterize the role of nitric oxide (NO) and NO-derived species in mouse models of inflammatory bowel disease (IBD) and cancer. These studies will use 129/SvEv Recombinase-activating gene 2 (Rag-2) knockout (KO) mice and C57BL/6 T cell receptor (TCR) alphabeta KO mice. Rag-2 KO mice on a 129/SvEv background develop severe IBD and cancer when infected with H. hepaticus. Histopathologic lesions and the pattern of macrophage and neutrophil infiltration will be correlated with biomarkers for nitration, oxidation, and halogenation of DNA and proteins in Rag-2 KO mice. These biomarkers will be further validated by comparing disease in naive and effector CD45RB high T cell-bearing Rag-2 KO mice, and by modulating NO production with N-methylarginine (NMA). TCR alphabeta KO mice develop severe IBD when infected with H. hepaticus. IBD in TCRbeta KO mice, like in Rag-2 KO mice, is a model for Crohn's disease (CD) and features activated macrophages and abundant interferon-gamma (IFN) and tumor necrosis factor-alpha (TNF). TCR alpha KO mice are a model for ulcerative colitis (UC), and are dependent on interleukin-4 (K,-4) but not IFN or TNF for the development of disease. Histopathologic lesions and cytokine levels will be correlated with biomarkers in TCR alpha and TCR beta KO mice. To further characterize the role of NO and NO-derived species in IBD, we will compare pharmacologic inhibition of inducible nitric oxide synthase (iNOS) with genetic inactivation of the enzyme by generating iNOS-deficient TCR alphabeta KO mice. In vitro analysis of somatic mutations arising in vivo in the presence and in the absence of NO and NO-derived species will also be performed. By using a novel IBD associated cancer model and models for UC and CD, we will be able to validate new biomarkers for nitration, oxidation, and halogenation of DNA and proteins. We will also gain a better understanding of the role of NO and NO-derived species across a broad range of carcinogenic events, including genotoxicity, cellular proliferation, cytotoxicity, and angiogenesis. The Specific Aims are: Specific Aim #1. Characterize the role of NO an oxidative stress in IBD and cancer in Rag-2 KO mice Specific Aim #2. Characterize the role of NO and oxidative stress in IBD in TCR alphabeta KO mice

该项目将描述一氧化氮 (NO) 和一氧化氮衍生物质在炎症性肠病 (IBD) 和癌症小鼠模型中的作用。这些研究将使用 129/SvEv 重组酶激活基因 2 (Rag-2) 敲除 (KO) 小鼠和 C57BL/6 T 细胞受体 (TCR) Alphata KO 小鼠。 129/SvEv 背景的 Rag-2 KO 小鼠在感染肝螺杆菌时会出现严重的 IBD 和癌症。 Rag-2 KO 小鼠的组织病理学病变以及巨噬细胞和中性粒细胞浸润的模式将与 DNA 和蛋白质硝化、氧化和卤化的生物标志物相关。这些生物标志物将通过比较原始小鼠和效应 CD45RB 高 T 细胞 Rag-2 KO 小鼠的疾病，并通过用 N-甲基精氨酸 (NMA) 调节 NO 产生来进一步验证。 TCR Alphata KO 小鼠在感染肝螺杆菌后会出现严重的 IBD。 TCRbeta KO 小鼠中的 IBD 与 Rag-2 KO 小鼠一样，是克罗恩病 (CD) 的模型，具有激活的巨噬细胞和丰富的干扰素-γ (IFN) 和肿瘤坏死因子-α (TNF)。 TCR α KO 小鼠是溃疡性结肠炎 (UC) 的模型，疾病的发生依赖于白细胞介素 4 (K,-4)，而不是 IFN 或 TNF。组织病理学病变和细胞因子水平将与 TCR α 和 TCR β KO 小鼠的生物标志物相关。为了进一步表征 NO 和 NO 衍生物种在 IBD 中的作用，我们将通过生成 iNOS 缺陷的 TCR Alphata KO 小鼠，比较诱导型一氧化氮合酶 (iNOS) 的药理学抑制与该酶的基因失活。还将对在存在和不存在NO和NO衍生物种的情况下体内产生的体细胞突变进行体外分析。通过使用新型 IBD 相关癌症模型以及 UC 和 CD 模型，我们将能够验证 DNA 和蛋白质硝化、氧化和卤化的新生物标志物。我们还将更好地了解 NO 和 NO 衍生物种在广泛的致癌事件中的作用，包括基因毒性、细胞增殖、细胞毒性和血管生成。具体目标是：具体目标#1。表征 Rag-2 KO 小鼠中 NO 和氧化应激在 IBD 和癌症中的作用具体目标#2。表征 NO 和氧化应激在 TCR Alphata KO 小鼠 IBD 中的作用