Gene Expression Profiling on Insulin Resistance and Obes

胰岛素抵抗和肥胖的基因表达谱

• 批准号: 6532223
• 负责人: Clifton Bogardus
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6532223
• 关键词: Native Americans; adipocytes; clinical research; differential display technique; gene expression; genetic regulation; genetic susceptibility; genome; human genetic material tag; human subject; hyperinsulinism; insulin sensitivity /resistance; linkage mapping; microarray technology; muscle cells; muscle metabolism; myoblasts; myotubes; obesity; polymerase chain reaction; striated muscles; tissue /cell culture

To identify novel genes that may contribute to insulin resistance, we examined skeletal muscle gene expression profiles in non-diabetic full-blooded Pima Indians using 1) human oligonucleotide microarrays containing 6800 genes and 35000 expressed sequence tags (ESTs), and 2) differential display PCR. We have also performed gene expression profiling in skeletal muscle and adipocyte samples from selected insulin resistant and insulin sensitive Swedish subjects in collaboration with Dr. Ulf Smith (Sahlgrenska University Hospital, Sweden)and Dr. Samuel Cushman (NIDDK). The potential roles of the differentially expressed genes/ESTs in insulin resistance will be evaluated by examining 1) their functions in relevant metabolic pathways, and additionally for Pima samples: 2) their chromosomal locations relative to peaks of linkage to insulin action and diabetes in a genome-wide scan in the Pima population. Skeletal muscle expression levels of selected genes were further investigated using real-time PCR. The manuscript on this project has been submitted. Previous studies have indicated strong correlations between biochemical characteristics of primary cultured skeletal muscle cells and clinical parameters of the donors. To determine which of the differentially expressed genes in muscle tissue of insulin resistant vs. insulin sensitive subjects may be primary, rather than secondary abnormalities, we propose also to compare the gene profiles of the cognate cultured myoblasts and myotubes. We have started to perform similar experiments to identify obesity susceptibility genes using freshly isolated adipocytes and cultured pre-adipocytes from lean and obese Pima Indians. Using oligonucleotide microarrays, we compared the expression profiles of adipocytes from 10 lean vs. 10 obese Pima Indians. We are analyzing the current data and collecting more samples for this project.

为了鉴定可能导致胰岛素抵抗的新型基因，我们使用1）使用1）含有6800个基因的人类寡核苷酸微阵列检查了非糖尿病全血PIMA印第安人中的骨骼肌基因表达谱，并且具有35000个基因和35000个表达的序列标签（ESTS），以及2）差异显示PCR。我们还与与瑞典Sahlgrenska大学医院的Ulf Smith博士和Samuel Cushman博士（NIDDK）合作，在骨骼肌和脂肪细胞样品中进行了基因表达分析。差异表达的基因/EST在胰岛素抵抗中的潜在作用将通过检查1）1）它们在相关代谢途径中的功能，以及PIMA样品的额外作用：2）相对于与胰岛素链接到胰岛素作用的峰值和PIMA中基因组全基因组扫描的染色体位置。使用实时PCR进一步研究了选定基因的骨骼肌表达水平。该项目的手稿已提交。先前的研究表明，原发培养的骨骼肌细胞的生化特征与供体的临床参数之间的相关性很强。为了确定胰岛素耐药性和胰岛素敏感受试者的肌肉组织中哪些差异表达的基因可能是主要的，而不是次级异常，我们还建议比较含同源培养的肌细胞和肌管的基因谱。我们已经开始执行类似的实验，以鉴定出新的分离的脂肪细胞和来自瘦肉和肥胖皮马印第安人的培养的脂肪细胞的肥胖敏感性基因。使用寡核苷酸微阵列，我们比较了10个瘦肉与10个肥胖皮马印第安人的脂肪细胞的表达谱。我们正在分析当前数据并为该项目收集更多样本。