COMPREHENSIVE GENE EXPRESSION PROFILING FOR TOXICOLOGY

分子毒理学研究，特别是研究化学对基因表达的影响的研究，提供了对毒性的许多机制的深入了解，并有助于评估环境中化学品的潜在健康风险。本提案的目的是进行和评估高通量基因表达测定，以监测细胞对潜在危险（有毒）化学物质的反应。在I期研究中，新型质量标记探针与质谱技术的结合被证明能够准确、灵敏地检测哺乳动物细胞中特定基因表达水平的变化。在第二阶段，该检测将作为一个高通量、自动化的系统实施，能够在每天数千个处理过的样品中评估对多个基因（10-25）的影响。基因将根据其与潜在公司合作伙伴和NIEHS感兴趣的毒理学终点的关联进行选择。除了基因选择外，该项目还将探索剂量反应分析、时间过程和不同细胞类型的使用在多大程度上提供了全面的表达谱。对这些特征的分析可用于将新化合物与已知化合物进行比较，以获得预测价值并更好地了解毒理学机制。拟议的商业应用：GeneTrace打算提供合同研究项目，使用本提案中描述的技术分析基因表达的化学效应。如果该分析是高通量自动化的，并且每个样品的成本足够低，则有很大的商业应用潜力。这些信息对毒理学家来说是很重要的，可以根据在不同浓度和时间内不同细胞类型中特定基因的诱导来对化学品进行分类。由于现代化学化合物库的规模，如果该分析是有效的，那么大型项目将对农业和化学工业的公司具有吸引力。

Studies in molecular toxicology, particularly those investigating chemical effects on gene expression, provide insight into the many mechanisms of toxicity and contribute to the assessment of potential health risks of chemicals in the environment. The objective of this proposal is to conduct and evaluate high-throughput gene expression assays for monitoring cellular responses to potentially hazardous (toxic) chemicals. in Phase I, the use of novel mass-labeled probes in conjunction with mass spectrometry was shown to be feasible for the accurate and sensitive detection of changes in the expression levels of selected sets of genes in mammalian cells. In Phase II, the assay will be implemented as a high- throughput, automated system capable of evaluating effects on multiple genes (10-25) in thousands of treated samples per day. Genes will be chosen for their association with toxicological endpoints that are of interest to potential corporate partners and to NIEHS. In addition to gene selection, the project will explore the extent to which analysis of dose response, time courses, and the use of different cell types provide comprehensive expression profiles. Analysis of these profiles may then be used for the comparison of novel against known compounds for predictive value and greater understanding of toxicological mechanisms. PROPOSED COMMERCIAL APPLICATION: GeneTrace intends to offer contract research programs for the analysis of chemical effects on gene expression using the technology described in this proposal. There is significant potential for commercial application if the assay is automated for high throughput and the cost per sample is sufficiently low. The information is important to toxicologists for classification of chemicals based on the induction of specific genes in a variety of cell types over a range of treatment concentrations and time. Because of the size of the modern chemical compound libraries, if the assay is efficient, then large-scale projects will be attractive to companies in the agricultural and chemical industries.