In Vivo Role of Nitric Oxide in Muscosal Inflammation and Cancer

项目4为了研究炎症、氧化应激和一氧化氮特别是导致癌症风险的机制，我们开发了一系列结肠炎和结肠炎相关癌症的小鼠模型。我们将在129/SvEv背景下使用白细胞介素10（IL 10）和重组激活基因2（Rag 2）缺陷的高度易感复合突变小鼠。这些129/SvEv IL 10 [-/-]Rag[-/-]小鼠在感染肝螺杆菌或过继转移CD 4 * CD 45 RB [高]效应T细胞（Teff）后发生类似炎症性肠病（IBD）和癌症的病变。我们还将使用感染幽门螺杆菌的C57 BL/6背景的IL 10 [-/-]小鼠和感染啮齿类柠檬酸杆菌的野生型C57 BL/6小鼠。对于所有三种小鼠模型，gpt δ报告基因转基因将用于定量和表征体内出现的体细胞突变。还将产生新的荧光黄色直接重复序列（FYDR）转基因，以定量体内重组事件。我们将描述诱导型一氧化氮合酶（iNOS）在感染H. C57 BL/6 IL 10 [-/-]小鼠感染H. trogontum感染IBD的C57 BL/6小鼠和C.会发展成自限性结肠炎这将通过NOS酶的药理学抑制以及通过将我们的C57 BL/6模型与iNOS[-/-]小鼠杂交来实现。还将进行消耗实验以确定中性粒细胞和巨噬细胞在这些模型系统中的结肠炎和癌症中的作用。这些实验将为本计划中的其他项目提供样本，以验证计算模型，表征化学生物标志物，并确定DNA损伤和修复在诱变中的作用。通过确定炎症细胞和大肠上皮细胞产生的一氧化氮的作用，我们将更好地了解IBD和一般粘膜炎症中癌症风险的机制基础。这些研究以及与该计划中其他项目的相互作用应转化为新的策略，以预防炎症相关癌症的发生或延迟其进展。

Project 4 In order to investigate the mechanisms by which inflammation, oxidative stress, and nitric oxide in particular contribute to cancer risk, we have developed a series of mouse models of colitis and colitis-associated cancer. We will use highly susceptible compound mutant mice deficient for interleukin 10 (IL10) and recombination activating gene 2 (Rag2) on a 129/SvEv background. These 129/SvEv IL10[-/-]Rag[-/-] mice develop lesions that resemble inflammatory bowel disease (IBD) and cancer when infected with Helicobacter hepaticus or after adoptive transfer with CD4*CD45RB[high] effector T cells (Teff). We will also use IL10[-/-] mice on a C57BL/6 background infected with Helicobacter trogontum and wild type C57BL/6 mice infected with Citrobacter rodentium. For all three mouse models, the gpt delta reporter transgene will be used to quantify and characterize somatic mutations that arise in vivo. A novel Fluorescent Yellow Direct Repeat (FYDR) transgene will also be generated in order to quantify recombination events in vivo. We will characterize the role of inducible nitric oxide synthase (iNOS) in 129/SvEv IL10[-/-]Rag[-/-] mice infected with H. hepaticus that develops a rapid onset of colitis-associated cancer, C57BL/6 IL10[-/-] mice infected with H. trogontum that develops IBD, and C57BL/6 mice infected with C. rodentium that develops a self-limiting colitis. This will be accomplished by pharmacologic inhibition of NOS enzymes as well as by crossing our C57BL/6 models with iNOS[-/-] mice. Depletion experiments will also be carried out to ascertain the role of neutrophils and macrophages in colitis and cancer in these model systems. These experiments will provide specimens for the other Projects in this Program in order to validate computational models, characterize chemical biomarkers, and establish the role of DNA damage and repair in mutagenesis. By establishing the role of nitric oxide produced by inflammatory cells and by epithelial cells in the large intestine, we will gain a better understanding of the mechanistic basis of cancer risk in IBD, and in mucosal inflammation in general. These studies and the interactions with the other Projects in the Program should translate into new strategies to prevent the initiation or delay the progression of inflammation-associated cancer.