Novel biosensors for toxicological applications

描述(由申请人提供)： 毒理基因组学已经成为替代缓慢、昂贵和低效的传统毒理学方法的一种彻底的替代方法。毒物基因组学方法是基于这样一个前提，即暴露在有毒物质中会引起独特的基因表达模式，可以通过基因阵列技术进行评估。然而，事实证明，分析数万个基因的表达模式是具有挑战性的。基因表达在很大程度上是在转录水平上调节的，并受转录因子(转录因子)控制，转录因子是结合调节基因元件(启动子和增强子)的蛋白质。据估计，存在数百个不同的转录因子家族，它们控制着由人类基因组编码的大约35,000个基因的表达。由于细胞调控的复杂性在转铁蛋白水平上大大降低，分析在转铁蛋白水平上发生的分子变化将为测试化合物的生物效应提供更简单和更全面的解释。Attagene Inc.已经开发出一种适用于分析多个转录因子活性的多转录因子系统(MTFS)。这项技术的可行性已在我们的初步研究中得到证明。在这里，我们将利用MTFS技术开发重组细胞系，作为各种毒理学应用的生物传感器。基于MTFS的测试系统的一个独特特征是模块化设计，可以组装许多MTF以评估任何组合的MTF。此外，模块化设计允许对现有的MTFS进行简单和快速的升级。在第一阶段的研究中，我们将通过构建一个原型测试系统来建立基于MTFS的生物传感器的方法学，该系统适用于确定几个控制对外源物质、毒物和应激反应的关键TF(例如PXR、PPARa、AhR、P53和NF-kB)。我们将测试这个系统的效用，以分析几种特性良好的毒物的毒性。最后，我们将评估基于MTFS的生物传感器的长期性能。在第二阶段的研究中，我们将推进我们的技术，使毒物学家能够获得新型生物传感器。通过允许分析几乎无限数量的TF，MTFS技术将毒物基因组学方法扩展到新的水平，从而为毒理学研究提供了一种独特的工具。

DESCRIPTION (provided by applicant): Toxicogenomics has emerged as a radical alternative to the slow, expensive and inefficient conventional toxicological approaches. The toxicogenomic approach is based on the premise that exposure to toxic agents elicits unique patterns of gene expression that can be assessed by using gene array technology. However, analyzing expression patterns of tens of thousands genes has been proven challenging. Gene expression is largely regulated on the transcriptional level and is controlled by transcription factors (TFs), proteins that bind regulatory gene elements (promoters and enhancers). An estimated few hundred of distinct TFs families exist that control the expression of approximately 35,000 genes encoded by the human genome. As the complexity of cell regulation is dramatically reduced at the TF level, analyzing the molecular changes occurring at the TF level would provide a much simpler and more comprehensive interpretation of biological effects of tested compounds. ATTAGENE Inc. has developed a multi-transcription factor system (MTFS) suitable for profiling the activities of multiple TFs. The feasibility of this technology has been demonstrated in our preliminary studies. Here, we will utilize the MTFS technology to develop recombinant cell lines that will serve as biosensors for a variety of toxicological applications. A unique feature of the MTFS-based test systems is a modular design that affords assembling many MTFS for assessment of any combination of TFs. Furthermore, the modular design allows for a simple and rapid upgrade of existing MTFSs. In the Phase I research, we will establish the methodology of the MTFS-based biosensors by constructing a prototypical test system suitable for determination of several key TFs controlling the responses to xenobiotics, toxicants, and stress (e.g. PXR, PPARa, AhR, p53, and NF-kB). We will test the utility of this system for profiling the toxicities of a few well-characterized toxicants. Finally, we will assess a long-term performance of the MTFS-based biosensors. In the Phase II research, we will advance our technology to the point where the novel biosensors can be made available for toxicologists. By allowing analyzing practically unlimited numbers of TFs, the MTFS technology expands the toxicogenomic approach to the new level and thus provides a unique tool for toxicological studies.