HDL GENE DISCOVERY--GENOME WIDE EXPRESSION SCREENS

HDL 基因发现——全基因组表达筛选

• 批准号: 6499032
• 负责人: EDWARD M RUBIN
• 金额: $ 45.7万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-05 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6499032
• 关键词: adrenal glands; apolipoproteins; blood lipoprotein metabolism; complementary DNA; enzyme activity; gene expression; gene mutation; genetic screening; genetically modified animals; genome; hepatic lipase; high density lipoproteins; laboratory mouse; lipid metabolism; liver; messenger RNA; microarray technology; phosphatidylcholine sterol acyltransferase

Based on the incompleteness of our understanding of High Density Lipoprotein (HDL) metabolism the focus of this proposal is the identification of new genes involved in the metabolism of this lipoprotein through genome-wide expression screens. Mouse cDNA arrays containing >5000 mouse genes will be used to identify genes whose expression is altered in the liver and adrenals of several transgenic and knockout lines of mice chosen based on their characterized abnormalities in HDL metabolism. This will initially include transgenic and knockout mice for apolipoprotein A-I (apo A-I), Scavenger Receptor class b1 (sr- bi), Hepatic lipase (HL), and Lecithin Cholesterol Acyl Transferase (LCAT). A basic assumption in these studies is that alterations in the expression of genes known to be involved in HDL metabolism will affect the expression of other genes also participating in the metabolism of this lipoprotein. The novel genes identified from these studies will be prioritized for further biological characterization based on a variety of parameters including: level of expression change, clustering of expression patterns between mice of different HDL mutant genotypes, and sequence or expression pattern similarities to other genes known to participate in lipoprotein metabolism. The function of a limited number of novel "HDL candidate" genes (approximately 10) will be assessed each year through their over-expression in transgenic mice coupled with careful analysis of the consequence of transgene over-expression over-expression on lipoprotein metabolism. In these studies we will be utilizing a combination of new technologies and previously developed experimental substrates to address the fundamental question of what genes are directly or indirectly involved in the metabolism of HDL in vivo.

基于我们对高密度脂蛋白（HDL）代谢的理解的不完整性，该提案的重点是鉴定了通过全基因组表达筛选的这种脂蛋白代谢的新基因。含有> 5000个小鼠基因的小鼠cDNA阵列将用于识别其表达在肝脏中改变的基因，并根据其根据HDL代谢中的特征异常而选择的几只转基因和基因敲除线的肾上腺。最初，这将包括载脂蛋白A-I（APO A-I），清道夫受体B1（SR- BI），肝脂肪酶（HL）和卵磷脂胆固醇酰基转移酶（LCAT）的转基因和基因敲除小鼠。这些研究中的一个基本假设是，与HDL代谢有关的基因表达的改变将影响其他基因的表达，也参与该脂蛋白的代谢。根据各种参数，将优先考虑从这些研究中发现的新基因，以进一步的生物学特征，包括：表达变化水平，不同HDL突变基因型的小鼠之间表达模式的聚类以及与参与脂蛋白代谢的已知的其他基因的序列或表达模式相似性。每年将通过在转基因小鼠中的过表达来评估有限数量的新型“ HDL候选”基因（约10个）的功能，并仔细分析转基因过表达对脂蛋白代谢的过表达的后果。在这些研究中，我们将利用新技术和以前开发的实验底物的组合来解决哪些基因直接或间接参与体内HDL代谢的基本问题。