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，并且与大血管并发症相结合时，死亡率很高。最常见的微血管并发症是神经病。尽管精确的患病率取决于用于鉴定神经病的诊断标准，但大多数研究表明，有50％的1型或2型糖尿病患者患有神经病。 通常认为，I型糖尿病患者所证明的循环葡萄糖的较大变化有助于糖尿病神经病的发展，而葡萄糖对基因表达的转录控制在糖尿病神经病的进展中起主要作用。尽管已经确定了肝细胞和胰腺β细胞中基因表达的葡萄糖调节的机制，但实际上对葡萄糖调节神经元基因表达的分子机制几乎一无所知。该建议将开发模型系统和新技术，以研究神经元消失表达的葡萄糖调节。这项工作将涉及费尔德曼和乌勒实验室之间的新合作。 三个特定的目的将用于集中提出的研究，该研究将采用两种神经母细胞瘤细胞系以及大鼠背根神经神经元在原发性培养中。在第一个特定目的中，新的高通量转染程序（表面转染和表达方案或步骤）将适用于神经母细胞瘤细胞系和原发性神经元。在第二个特定目的中，将使用DNA微阵列杂交鉴定由葡萄糖转录调节的候选基因。在第三个特定目的中，将使用步骤转染研究这些候选基因的启动子以及其他特征良好的启动子元素的调节。 研究的完成不仅将确定导致糖尿病神经并发症发展的关键基因组机制，而且还将为开发药物筛查测定法的基础奠定基础，这对于测试药物以干扰这些机制至关重要。