Development of in vitro toxicity expression assays

描述(由申请人提供)： 这项提案的目标是开发和验证一种用于毒理学的体外基因表达分析方法。具体地说，我们建议使用体外大鼠原代肝细胞的全基因组表达谱来获得可以根据特定的肝脏病理终点对化学物质进行分类的基因签名，并将基因签名转换为高效、高通量、有效的分析方法，作为一种经济高效、快速的筛选工具。该项目将利用ICONIX研究人员在使用复合治疗和全球微阵列基因表达分析识别毒理学基因特征方面的联合专业知识，以及Althea研究人员将这些特征转化为使用高度多路复用、通用启动的RTPCR(UP-RTPCR)的高通量基因表达分析的联合专业知识。在这个第一阶段的提案中，我们打算演示一个普通的肝毒性终点，胆固酮，和一个普通毒性终点，DNA损伤的过程。

DESCRIPTION (provided by applicant): The goal of this proposal is to develop and validate a process for generating in vitro, gene expression assays for toxicology. Specifically, we propose to derive gene signatures, that can classify chemicals according to specific liver pathology endpoints, using whole genome expression profiling in rat primary hepatocytes in vitro and to convert the gene signatures into efficient, high throughput, validated assays for use as a cost efficient, fast screening tool. The project will utilize the joint expertise of Iconix researchers in identifying toxicological gene signatures using compound treatment and global, microarray gene expression analysis, and Althea researchers for translating these signatures into high throughput gene expression assays using highly multiplexed, universal-primed rtPCR (UP-rtPCR). In this Phase I proposal, we intend to demonstrate the process for one general hepatoxicity endpoint, Cholestatis, and one general toxicity endpoint, DNA damage.