Gene Expression in Cancer by Microarray Hybridization

从一开始，该计划的目标就是开发和应用全基因组 癌症基因表达模式的系统和定量分析方法。在这八年里 自从我们开始这个项目以来，全球基因系统研究的实验和分析工具 转录丰度水平的表达模式已经戏剧性地发展起来，它们已经成为 广泛使用并广泛用于癌症研究。由此产生的研究已经产生了丰富的 对癌症的新见解，他们甚至开始影响病人的护理。分析mRNA转录本 然而，水平只提供了全球基因表达计划的部分情况。趁有机会 大量证据表明，对信使核糖核酸的翻译、亚细胞定位和衰退的调控至关重要 生物调控的要素，实用和强大的全基因组方法来研究这些水平的 监管仍有待制定。因此，我们对系统体系结构、分子 这些调节机制的机制和生物学作用，包括它们在人类中所起的作用 癌症，几乎还处于初级阶段。因此，我们建议开发实用、健壮、高吞吐量的方法 使用DNA微阵列描述转录后水平上全球调控的三个关键方面： 翻译调控、信使核糖核酸降解调控和RNA结合的特异性相互作用 蛋白质和它们的靶标。这些研究将建立在我们已经在 酿酒酵母模型系统，但将重点放在人类细胞上。作为基本的方法论 ，我们将应用它们来开发基本的知识框架，调查 这些调节机制在基本生理和发育中的应用模式 程序，它们如何从一种细胞类型到另一种细胞类型和个体之间的差异，并开始调查 潜在的分子机制。我们的目标是开发实验方法和 解释框架，以使这些转录后水平的基因表达 在全基因组范围内系统地分析和研究，几乎和转录水平一样例行公事 今天。

From the start, the goal of this program has been the development and application of genome-wide approaches to systematic and quantitative analysis of gene expression patterns in cancer. In the eight years since we began this project, the experimental and analytical tools for systematic studies of global gene expression patterns at the level of transcript abundance have developed dramatically and they have become widely available and widely used for the study of cancer. The resulting studies have produced a wealth of new insight into cancer and they are even beginning to influence patient care. Profiling mRNA transcript levels, however, provides only a partial picture of the global gene expression program. While there is abundant evidence that regulation of the translation, subcellular localization and decay of mRNA are critical elements of biological regulation, practical and robust genome-wide approaches to studying these levels of regulation remain to be developed. As a result, our knowledge of the systems architecture, molecular mechanisms and biological roles of these regulatory mechanisms, including the roles they play in human cancer, is barely in its infancy. We therefore propose to develop practical, robust, high-throughput methods using DNA microarraysto profile three critical aspects of global regulation at the post-transcriptional level: translational regulation, regulation of mRNA degradation, and the specific interactions of RNA binding proteins with their targets. These studies will build on preliminary studies we've already begun in the Saccharomyces cerevisiae model system, but will focus on human cells. As the essential methodologies are developed, we will apply them to developing a foundational framework of knowledge, investigating the patterns in which these regulatory mechanisms are used in basic physiological and developmental programs, how they vary from one cell type to another and between individuals, and beginning to investigate the underlying molecular mechanisms. The goal is to develop both the experimental methodology and an interpretive framework to the point that these post-transcriptional levels of gene expression can be systematically profiled and studied on a genome-wide scale almost as routinely as transcript levels are today.