Project 2: Functional Profiling of Susceptibility Genes

人类对超级基金网站发现的化学物质的有毒影响的遗传易感性各不相同。 癌症和其他形式的毒性可能是由不利的基因-环境相互作用引起的。遗传 对一种化学物质毒性作用的敏感性可能与该化学物质的细胞靶标有关。 及其代谢物。然而，我们对许多细胞靶点的了解仍然有限。 超级基金名单上的优先化学品。我们将利用基本代谢守恒的优势 酿酒酵母与人类之间的途径和基本细胞过程 候选人类易感基因。在这里，我们提出了一种在酵母中发现这些靶点的新方法。 和人类细胞分别使用平行缺失分析(PDA)和RNA干扰(RNAi)。删除 几乎每个酵母基因的菌株都使新的方法能够并行(PDA)确定亲缘关系 每个酵母基因对化学毒物敏感性的重要性。我们建议确定 选择需要代谢激活的优先超级基金化学品的候选易感基因 包括苯、多环芳烃(PAHs)、卤代脂肪族碳氢化合物以及 选定的金属，如砷和镉，不会。我们将选择可能的人类并对其进行优先排序 通过对酵母数据集的计算分析，得到候选基因。候选的人类易感性 在适当的人类细胞系中，使用RNAi将使基因沉默，从而使它们在敏感性方面的作用 超级基金化学物质和/或其代谢物的细胞毒性、遗传毒性和表观遗传效应 已评估。我们认为，这种方法将识别使人类对超级基金易感的基因。 化学物质及其代谢物，并将使未来的工作，以检查变种之间的联系 这些基因和不良后果。此外，这项工作可能会为 导致超级基金优先化学品毒性的细胞过程。

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.