Massively Parallel Gene Expression Analysis

大规模并行基因表达分析

• 批准号: 6413039
• 负责人: RICHARD J. QUIGG
• 金额: $ 51.88万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-25 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6413039
• 关键词: animal tissue; biotechnology; computer data analysis; cooperative study; diagnosis design /evaluation; disease /disorder classification; disease /disorder model; endocrine disorder; gastrointestinal disorder; gene expression; genetic mapping; high throughput technology; human tissue; informatics; kidney disorder; microarray technology; subtraction hybridization; technology /technique development

The research proposed in this application will develop and utilize massively parallel gene analysis technology. Both commercially available microarrays from Affymetrix and arrays custom-made in this facility will be used. The basic strategy for these experiments is to identify genes that are relevant for individual experimental projects. These finite gene sets will be included on arrays made in our facility, which will be customized for the individual user. The advantages of such an approach are that multiple experiments will then be performed cost effectively, allowing appropriate statistical analyses of data, as well as systematic variations and manipulations. Ongoing studies are those from NIDDK-funded investigators examining disease processes and models in humans, animals and homogenous cells: 1) Nephropathy occurring during the course of experimental diabetes mellitus in mice; 2) Lymphocytic abnormalities in mice with experimental inflammatory bowel disease (IBD); 3) Molecular basis of intestinal epithelial adaptation to altered luminal sodium load; 4) Activation defects in monocytes from patients with IBD; 5) Effects of steroids on muscle and adipocyte gene expression in polycystic ovary syndrome; 6) Analysis of signaling defects in mouse models of thyroid hormone resistance; 7) Analysis of signaling defects in patients with thyroid hormone resistance; 8) Analyses of mRNA changes in human pancreatic islets prepared for transplantation; 9) Effects of high glucose exposure to pancreatic beta cells; and, 10) Neuronal gene expression in response to gonadotropin-releasing hormone. These analyses will permit the identification of relevant genes that are upregulated or downregulated in these various conditions. Pilot and feasibility studies will be to: 1) Develop cDNA microarrays from pituitary, pancreatic islets and renal glomeruli to allow gene profiling from these anatomically unique sites; and 2) Utilize laser capture microdissection microscopy to obtain glomerular RNA from animal models and human tissue specimens of lupus nephritis, and subsequently apply microarray technology to identify gene changes in this disease. If this NIDDK-sponsored core facility is funded, innovative work investigating gene expression in a variety of renal, endocrinologic and gastrointestinal disease models and processes will be performed.

本应用程序中提出的研究将开发和利用大量平行的基因分析技术。将使用来自Affymetrix的市售微阵列和该设施中定制的阵列。这些实验的基本策略是确定与单个实验项目相关的基因。这些有限的基因集将包含在我们的设施中制作的阵列中，该阵列将为单个用户定制。这种方法的优点是，将有效地进行多个实验，从而可以对数据进行适当的统计分析以及系统的变化和操纵。正在进行的研究是来自NIDDK资助的研究人员检查人类，动物和同质细胞中疾病过程和模型的研究者：1）在小鼠实验性糖尿病过程中发生的肾病； 2）具有实验性炎症性肠病（IBD）的小鼠的淋巴细胞异常； 3）肠上皮上皮适应改变腔钠负荷的分子基础； 4）来自IBD患者的单核细胞的激活缺陷； 5）类固醇对多囊卵巢综合征肌肉和脂肪细胞基因表达的影响； 6）分析甲状腺激素耐药性小鼠模型中的信号缺陷； 7）分析甲状腺激素耐药性患者的信号缺陷； 8）分析准备移植的人类胰岛变化； 9）高葡萄糖暴露于胰腺β细胞的影响； 10）响应促性腺激素释放激素的神经元基因表达。这些分析将允许在各种条件下上调或下调的相关基因鉴定。试点和可行性研究将是：1）从垂体，胰岛和肾肾小球中发展cDNA微阵列，以允许从这些解剖学上独特的位点进行基因分析； 2）利用激光捕获显微镜显微镜从狼疮肾炎的动物模型和人体组织标本中获得肾小球RNA，然后应用微阵列技术来识别该疾病中的基因变化。如果为NIDDK赞助的核心设施提供资金，将进行各种肾脏，内分泌和胃肠道疾病模型和过程中研究基因表达的创新工作。