Project 2: Functional Profiling of Susceptibility Genes

人类对超级基金场所发现的化学物质的毒性影响的遗传易感性各不相同。 不利的基因与环境相互作用可能会导致癌症和其他形式的毒性。遗传 对化学品毒性作用的敏感性可能与化学品的细胞靶点有关 及其代谢物。然而，我们对许多肿瘤的细胞靶点的了解仍然有限。 超级基金名单上的优先化学品。我们将利用基础代谢的保守性 途径和基本的细胞过程之间的酵母S。酿酒酵母和人类来鉴定 候选人类易感基因。在这里，我们提出了一种新的方法来发现这些目标在酵母 和人细胞，分别使用平行缺失分析（PDA）和RNA干扰（RNAi）。删除 几乎每一个酵母基因的菌株使新的方法来确定平行（PDA）的相对 每个酵母基因对化学毒物敏感性的重要性。我们建议确定 需要代谢活化的选定优先超级基金化学品的候选易感基因 包括苯、多环芳烃（PAH）、卤代脂族烃，以及 选择的金属，如砷和镉，不。我们将选择和优先考虑可能的人类 候选基因的酵母数据集的计算分析。候选人人类易感性 基因将在适当的人类细胞系中使用RNAi沉默，使得它们在对 超级基金化学品和/或其代谢物的细胞毒性、遗传毒性和表观遗传效应可 评估。我们认为，这种方法将确定基因赋予人类易感性的超级基金 化学物质及其代谢物，并将使未来的工作，以检查之间的关联变异， 这些基因和不良后果。此外，这项工作可能会提供重要的见解， 细胞过程导致超级基金优先化学品的毒性。

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.