Postgenomic approaches to diabetic complications

糖尿病并发症的后基因组学方法

• 批准号: 6666784
• 负责人: MICHAEL D UHLER
• 金额: $ 33.98万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6666784
• 关键词: apoptosis; biological signal transduction; diabetes mellitus genetics; diabetic neuropathy; disease /disorder model; experimental designs; gene expression; genetic promoter element; genetic regulation; genetic transcription; glucose; high throughput technology; laboratory rat; microarray technology; model design /development; neuroblastoma; neuroregulation; spinal ganglion; technology /technique development; tissue /cell culture; transfection

DESCRIPTION (provided by applicant): The microvascular complications of type 1 diabetes carry a high morbidity and, when coupled with macrovascular complications, high mortality. The most common microvascular complication is neuropathy. While exact prevalence depends on the diagnostic criteria used to identify neuropathy, most studies suggest that 50% of patients with either type 1 or type 2 diabetes have neuropathy. It is generally assumed that the large variations in circulating glucose evidenced in type I diabetics contribute to the development of diabetic neuropathy and that transcriptional control of gene expression by glucose plays a major role in the progression of diabetic neuropathy. Although mechanisms have been identified for glucose regulation of gene expression in hepatocytes and pancreatic beta cells, virtually nothing is know about the molecular mechanism by which glucose regulates gene expression in neurons. This proposal will develop model systems and new technologies for the study of glucose regulation of neuronal gone expression. This work will involve a new collaboration between the Feldman and Uhler laboratories. Three specific aims will serve to focus the proposed studies which will employ two neuroblastoma cells lines as well as rat dorsal root ganglion neurons in primary culture. In the first specific aim, a new high-throughput transfection procedure (Surface Transfection and Expression Protocol or STEP) will be adapted to the neuroblastoma cell lines and primary neurons. In the second specific aim, candidate genes transcriptionally regulated by glucose will be identified using DNA microarray hybridization. In the third specific aim, the regulation of promoters from these candidate genes as well as other well-characterized promoter elements by glucose will be studied using STEP transfection. The completion of the studies will not only identify critical genomic mechanisms responsible for the development of neurological complications of diabetes, but they will lay the foundation for the development of drug screening assays that will be essential for the testing of drugs to interfere with these mechanisms.

描述（由申请人提供）： 1型糖尿病的微血管并发症具有高发病率，并且当与大血管并发症结合时，具有高死亡率。最常见的微血管并发症是神经病变。虽然确切的患病率取决于用于识别神经病变的诊断标准，但大多数研究表明，50%的1型或2型糖尿病患者患有神经病变。 一般认为，在I型糖尿病患者中证实的循环葡萄糖的大的变化有助于糖尿病神经病变的发展，并且葡萄糖对基因表达的转录控制在糖尿病神经病变的进展中起主要作用。尽管已经确定了葡萄糖调节肝细胞和胰腺β细胞中基因表达的机制，但实际上对葡萄糖调节神经元中基因表达的分子机制一无所知。本研究将为葡萄糖调节神经元Gone表达的研究开发模型系统和新技术。这项工作将涉及费尔德曼和乌勒实验室之间的新合作。 三个具体的目标将集中在拟议的研究，将采用两个神经母细胞瘤细胞系以及大鼠背根神经节神经元的原代培养。在第一个具体目标中，一种新的高通量转染程序（表面转染和表达协议或STEP）将适用于神经母细胞瘤细胞系和原代神经元。在第二个具体目标中，将使用DNA微阵列杂交来鉴定受葡萄糖转录调控的候选基因。在第三个具体目标中，将使用STEP转染研究葡萄糖对来自这些候选基因的启动子以及其他充分表征的启动子元件的调节。 这些研究的完成不仅将确定负责糖尿病神经系统并发症发展的关键基因组机制，而且还将为开发药物筛选试验奠定基础，这些试验对于检测干扰这些机制的药物至关重要。