Project 2: Oxidative DNA Damage and Genetic Instability In Models Of Intestinal

项目2：肠道模型中的氧化DNA损伤和遗传不稳定性

基本信息

批准号：
7511051
负责人：
Paul William Doetsch
金额：
$ 28.42万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7511051
关键词：
Address Agar Gel Electrophoresis Alkaline Single-Cell Gel Electrophoresis Assay Animals Attention Cancer Model Cell Culture System Cell Separation Cells Chromosome abnormality Chromosomes Chronic Colon Carcinoma Colonic Neoplasms DNA DNA Damage DNA Microarray Chip DNA Microarray format DNA Repair DNA Sequence Rearrangement DNA-Directed DNA Polymerase Development Diagnostic Neoplasm Staging Enzymes Excision Flow Cytometry Fluorescent Probes Fluorescent in Situ Hybridization Gel Genetic Genetic Recombination Genome Genomics Intestinal Cancer Intestinal Neoplasms Intestines Mammalian Cell Maps Mediating Mediator of activation protein Methodology Mitochondria Mitochondrial DNA Modeling Monitor Mus Mutation Nature Normal Cell Oxidative Stress Pathway interactions Personal Satisfaction Phenotype Physiological Point Mutation Reciprocal Translocation Recurrence Role Saccharomyces cerevisiae Series Stress System Tissues Tumor stage Yeasts biological adaptation to stress cancer cell comparative genomic hybridization design mouse model mutant oxidative DNA damage prevent programs repaired research study response stress management tumor

项目摘要

Oxidative DMA Damage and Genetic Instability in Models of Intestinal Tumor Development It is well-established that oxidative DMA damage can cause mutations that have been implicated in tumor development. During the previous program project support period, we exploited isogenic yeast strains to determine the relationships between those pathways involved in removal/repair of oxidative DMA damage and those pathways that tolerate its presence in the genome via DNA polymerase-mediated translesion synthesis and recombination leading to genetic instability. In addition, we have established that the chronic presence of unrepaired DNA damage causes a "stress adaptation" response that leads to an increase in intracellular ROS accompanied by further increases in genetic instability and a host of physiological changes similar to the cancer cell phenotype. In project 2, we will further define the relevance of the DNA damagemediated ROS/genetic instability response within the context of it being a driver of small- and large-scale genomic rearrangements in both yeast and mammalian cells. We will also determine the relationships among oxidative DNA damage, DNA repair and genetic instability activities during various stages of tumor development in a mouse colon cancer system where mitochondrial- and Nox-mediated generators of ROS have been activated. We will also continue our collaborative studies to further elucidate the connections between oxidative mtDNA damage, mtDNA genetic instability and those systems that respond (or fail to respond) to such damage. The specific aims of this project are designed to exploit yeast to rapidly identify key ROS mediators of genetic instability and to define the nature of abnormal chromosomal changes caused by ROS as a result of DNA damage caused by exogenous and endogenous agents. The results of the yeast studies will be used to focus attention on similar relationships that we hypothesize will exist during the development of mammalian colon tumors where increasing intracellular ROS functions as a driver (via oxidative DNA damage) of small- and large-scale genomic changes, from point mutations to chromosomal aberrations, including amplifications, deletions, and non-reciprocal translocations. The results of these studies should provide a picture of the extent to which ROS functions as the mediator of genetic instability during colon tumor development in mammalian intestinal cells.

肠道肿瘤发展模型中的氧化DMA损伤和遗传不稳定众所周知的是，氧化DMA损伤可能导致与肿瘤有关的突变发展。在上一个计划的项目支持期间，我们将等源性酵母菌菌株利用确定涉及去除/修复DMA损伤的途径之间的关系以及那些通过DNA聚合酶介导的转移耐受其在基因组中存在的途径合成和重组导致遗传不稳定性。此外，我们已经确定了慢性未修复的DNA损伤的存在会导致“应力适应”响应，导致增加细胞内ROS伴随着遗传不稳定性的进一步增加和许多生理变化与癌细胞表型相似。在项目2中，我们将进一步定义DNA Damagemed的相关性 ROS/遗传不稳定性反应在小规模和大规模的驱动器中酵母和哺乳动物细胞中的基因组重排。我们还将确定关系在肿瘤各个阶段的氧化DNA损伤中，DNA修复和遗传不稳定性活性在小鼠结肠癌系统中开发线粒体和NOX介导的ROS的发电机已被激活。我们还将继续我们的协作研究，以进一步阐明联系在氧化mtDNA损伤，mtDNA遗传不稳定性与响应的系统之间（或无法响应）这种损害。该项目的具体目的旨在利用酵母来快速识别关键的ROS介质遗传不稳定并定义由ROS引起的异常染色体变化的性质由外源和内源性剂引起的DNA损伤。酵母研究的结果将被使用将注意力集中在我们假设的类似关系上哺乳动物结肠肿瘤，其中增加细胞内ROS充当驱动器（通过氧化DNA 损害）小型和大型基因组变化，从点突变到染色体畸变，包括放大，缺失和非转换易位。这些研究的结果应该提供一张ROS在结肠过程中充当遗传不稳定性介体的程度哺乳动物肠细胞的肿瘤发展。