COMPREHENSIVE GENE EXPRESSION PROFILING FOR TOXICOLOGY

分子毒理学研究，特别是那些调查化学品对基因表达的影响的研究，有助于深入了解毒性的许多机制，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检测，用于监测细胞对潜在危险（有毒）化学品的反应。在第一阶段中，使用新型的质量标记探针结合质谱法被证明是可行的，可以准确和灵敏地检测哺乳动物细胞中选定的基因组的表达水平的变化。在第II阶段，该测定将作为高通量自动化系统实施，该系统能够评价每天数千个处理样品中对多个基因（10-25个）的影响。将选择与潜在企业合作伙伴和NIEHS感兴趣的毒理学终点相关的基因。除了基因选择之外，该项目还将探索剂量反应分析、时间进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。然后，这些特征的分析可用于比较新化合物与已知化合物，以获得预测值并更好地理解毒理学机制。拟定商业应用：GeneTrace打算提供合同研究计划，使用本提案中描述的技术分析化学品对基因表达的影响。如果该测定是自动化的，以获得高通量，并且每个样品的成本足够低，则商业应用有很大的潜力。这些信息对于毒理学家根据在一系列处理浓度和时间内诱导各种细胞类型中的特定基因对化学品进行分类非常重要。由于现代化合物库的规模，如果分析是有效的，那么大规模的项目将吸引农业和化学工业的公司。

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.