In Vivo Role of Nitric Oxide in Muscosal Inflammation and Cancer

项目4为了研究炎症、氧化应激、 而一氧化氮尤其会增加癌症的风险，我们已经开发出一系列小鼠 结肠炎和结肠炎相关癌症的模型。我们将使用高度敏感的复合突变体 白介素10(IL10)和重组激活基因2(RAG2)缺陷的小鼠 129/SvEv背景。这些129/SvEv ILIO‘^RAG“”小鼠会出现类似于 炎症性肠病(IBD)和感染肝螺杆菌或癌症 用CD4*CD45Rb高效性T细胞(T细胞)过继移植后。我们还将使用IL10“‘”小鼠 感染弓形螺杆菌的C57BL/6小鼠和野生型C57BL/6小鼠的研究 感染了轮状柠檬酸杆菌。对于所有三种鼠标模型，gpt Delta记者 转基因将被用来量化和描述体内出现的体细胞突变。一本小说 还将产生荧光黄色直接重复序列(FYDR)转基因以量化 体内重组事件。我们将描述诱导型一氧化氮合酶的作用 (INOS)在129/SvEv ILIO^‘rag“”感染His的小鼠中的作用 结肠炎相关癌症，感染传染性法氏囊虫的C57BL/6 IL10“”小鼠， 以及感染了齿状念珠菌的C57BL/6小鼠，这种小鼠会发展为自限性结肠炎。这将是 通过药物抑制一氧化氮合酶以及通过交叉我们的C57BL/6 INOS“‘”小鼠模型。还将进行耗竭实验，以确定 在这些模型系统中，结肠炎和癌症中的中性粒细胞和巨噬细胞。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，表征化学生物标记物，并确定DNA损伤和修复在 诱变。通过确定由炎性细胞产生的一氧化氮和通过 在大肠上皮细胞中，我们会对其机制有更好的了解 IBD和一般粘膜炎症的癌症风险的基础。这些研究和 与计划中其他项目的互动应转化为新的战略，以 预防或延缓炎症相关癌症的发生或进展。

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 ILIO'^Rag"'" mice develop lesions that resemble inflammatory bowel disease (IBD) and cancer when infected with Helicobacter hepaticus or after adoptive transfer with CD4*CD45RBhigh 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 ILIO^'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 newstrategies to prevent the initiation or delay the progression of inflammation-associated cancer.