Project 3: Cellular Effects of Aldehydic Products of Lipid Peroxidation

来自几个实验室的新证据表明，脂质氧化产物起主要作用 人类疾病。一类重要的脂质氧化产物是亲电体， 亲核试剂包括谷胱甘肽、蛋白质和DNA。虽然谷胱甘肽和DMA修饰已经被 经过多年的研究，不充分的技术限制了我们研究蛋白质修饰的能力， 与生物学结果的关系。蛋白质组学和基因组学的技术进步使其成为可能 组装由脂质亲电体修饰的蛋白质的全面概况以及转录 他们带来的变化。使用系统方法比较这些密集数据集， 将蛋白质修饰与细胞反应联系起来。跨世纪计划项目研究 在过去的两年中，已经产生了用于鉴定脂质电泳修饰的关键试剂和新方法。 蛋白质及其生物学后果。我们现在准备发起一场激进的运动 定义脂质亲电体修饰蛋白质的化学和生物学及其与 炎症是许多人类疾病的主要原因。项目3的作用是确定 从项目1和2的研究中产生的生物学上有趣的脂质亲电体，并定义 它们在人巨噬细胞系THP-1中诱导的转录变化。此信息将 与将在项目4中确定的相同试剂的蛋白质修饰模式相比， 以及将蛋白质修饰与生物学结果联系起来的假设。这些假设将是 使用诸如RNA敲低、异位蛋白表达和 生化分析这将有助于阐明脂质对蛋白质修饰的机制， 嗜电物质会导致促炎和抗炎反应。

Emerging evidence from several laboratories indicates that lipid oxidation products make major contributions to human disease. An important class of lipid oxidation products are electrophiles that react with cellular nucleophiles including glutathione, proteins, and DNA. Although glutathione and DMA modification has been studied for many years, inadequate technology has limited our ability to investigate protein modification and its relation to biological outcome. Technological advances in proteomics and genomics now make it possible to assemble comprehensive profiles of proteins modified by lipid electrophiles as well as the transcriptional changes they induce. Comparison of these dense datasets using systems approaches enables hypotheses to be generated linking protein modification to cellular response. Research in the Program Project over the past two years has generated critical reagents and novel approaches for identifying lipid electrophilemodified proteins and their biological consequences. We are now poised to mount an aggressive campaign to define the chemistry and biology of protein modification by lipid electrophiles and its relationship to inflammation, a major contributor to many human diseases. The role of Project 3 is to identify the most biologically interesting lipid electrophiles emanating from research in Projects 1 and.2 and to define the transcriptional changes that they induce in the human macrophage cell line, THP-1. This information will be compared to the pattern of protein modification by the same agents, which will be determined in Project 4, and hypotheses formulated linking protein modification to biological outcome. These hypotheses will be tested using a combination of approaches such as RNA knockdown, ectopic protein expression, and biochemical assays. This will enable the elucidation of mechanisms by which protein modifications by lipid lectrophiles lead to pro- and anti-inflammatory responses.