Project 2: Functional Profiling of Susceptibility Genes

项目2：易感基因的功能分析

• 批准号: 7089422
• 负责人: CHRISTOPHER D VULPE
• 金额: $ 31.2万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7089422
• 关键词: DNA damage; RNA interference; Saccharomyces cerevisiae; arsenic; benzene; bioinformatics; cadmium; carbopolycyclic compound; computational biology; cytotoxicity; environmental exposure; environmental toxicology; epigenetics; fungal genetics; gene deletion mutation; gene environment interaction; genetic susceptibility; halogenation; hydrocarbons; metabolism; molecular dynamics

Humans vary in their genetic susceptibility to the toxic effects of chemicals found at Superfund sites. Cancers and other forms of toxicity may arise from adverse gene-environment interactions. Genetic susceptibility to the toxic effects of a chemical is likely to be related to the cellular targets of the chemical and its metabolites. However, we still have a limited understanding of cellular targets for many of the priority chemicals on the Superfund list. We will take advantage of the conservation of basic metabolic pathways and fundamental cellular processes between the yeast S. cerevisiae and humans to identify candidate human susceptibility genes. Here, we propose a new approach to discover these targets in yeast and human cells using parallel deletion analysis (PDA) and RNA interference (RNAi), respectively. Deletion strains for almost every yeast gene enable new approaches to determine in parallel (PDA) the relative importance of each yeast gene for susceptibility (sensitivity) to a chemical toxicant. We propose to identify candidate susceptibility genes for selected priority Superfund chemicals that require metabolic activation including benzene, polycyclic aromatic hydrocarbons (PAHs), halogenated aliphatic hydrocarbons, and for selected metals such as arsenic and cadmium, which do not. We will select and prioritize likely human candidate genes by computational analysis of the yeast data sets. The candidate human susceptibility genes will be silenced in appropriate human cell lines using RNAi so that their roles in sensitivity to cytotoxicity, genotoxicity and epigenetic effects of the Superfund chemical and/or its metabolites can be evaluated. We suggest that this approach will identify genes that confer human susceptibility to Superfund chemicals and their metabolites and will enable future work to examine associations between variants in these genes and adverse outcomes. In addition, this work will likely provide important insights in the cellular processes leading to toxicity for priority Superfund chemicals.

人类对在超级基金部位发现的化学物质的毒性作用的遗传敏感性各不相同。 癌症和其他形式的毒性可能来自不良基因环境相互作用。遗传 对化学物质毒性作用的敏感性可能与化学物质的细胞靶标有关 及其代谢物。但是，我们仍然对许多细胞靶标的许多 超级基金列表中的优先化学品。我们将利用基本代谢的保护 酵母菌和人类之间的途径和基本细胞过程以识别 候选人类易感基因。在这里，我们提出了一种新的方法，以发现酵母中的这些靶标 分别使用平行缺失分析（PDA）和RNA干扰（RNAi）和人类细胞。删除 几乎每个酵母基因的菌株都可以使新方法并行确定相对（PDA） 每个酵母基因对化学有毒物质的敏感性（敏感性）的重要性。我们建议确定 需要代谢激活的选定优先超级基金化学物质的候选敏感性基因 包括苯，多环芳烃（PAHS），卤化脂肪液碳氢化合物，以及用于 选定的金属，例如砷和镉，不可能。我们将选择并确定可能的人 通过对酵母数据集的计算分析候选基因。候选人的易感性 基因将在适当的人类细胞系中使用RNAi沉默，从而使其在敏感性中的作用 超级基金化学和/或其代谢物的细胞毒性，遗传毒性和表观遗传学作用可以是 评估。我们建议这种方法将确定具有人类对超级基金敏感性的基因 化学物质及其代谢产物，并将使以后的工作能够检查变体之间的关联 这些基因和不良结果。此外，这项工作可能会在 细胞过程导致优先超级基金化学物质的毒性。