Gene Expression Profiling On Insulin Resistance And Obes

我们进行了全球基因表达谱分析，以补充正在进行的定位克隆工作，以确定糖尿病及其预测因素、胰岛素抵抗和肥胖症的易感基因。为此，我们使用了1)包含6,800个基因和35000个表达序列标签(EST)的人类寡核苷酸微阵列和2)差异显示聚合酶链式反应(Differential Display PCR2)，随后采用实时RT-PCR法检测了非糖尿病全血PIMA印第安人的骨骼肌基因表达谱。这项研究发表在《糖尿病》杂志上，DOI 10.1007/s00125002-0905-7。通过检测1)差异表达基因/EST在相关代谢途径中的功能，例如IRS-1(糖尿病51，补充材料2：A266)，以及2)它们在全基因组扫描中与胰岛素作用和糖尿病相关的峰值的染色体位置，例如ORP150(糖尿病51：1618-1621)，评估差异表达基因/EST在胰岛素抵抗中的潜在作用。 我们还将使用原代人类骨骼肌培养系统来鉴定潜在的胰岛素信号通路新成员，并比较来自相同捐赠者的人类肌肉活检组织和培养的肌肉细胞之间的基因图谱。 为了确定肥胖的易感基因，我们使用20名精瘦和19名肥胖的皮马印第安人的新鲜分离的脂肪细胞进行了基因表达谱实验。我们目前正在分析这些数据，并将根据与肥胖指标相关联的功能和染色体位置来寻找潜在的肥胖易感基因。

We performed global gene expression profiling to complement the ongoing positional cloning effort to identify susceptibility genes for diabetes and its predictors, insulin resistance and obesity. To this end, we have 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, followed by Real Time RT-PCR. This work has been published in Diabetologia, DOI 10.1007/s00125-002-0905-7. The potential roles of the differentially expressed genes/ESTs in insulin resistance are evaluated by examining 1) their functions in relevant metabolic pathways, such as IRS-1 (Diabetes 51, Supplement 2: A266) and 2) their chromosomal locations relative to peaks of linkage to insulin action and diabetes in a genome-wide scan in the Pima population, such as ORP150 (Diabetes 51:1618-1621). We will also use primary human skeletal muscle culture system to identify potentially novel members of insulin signaling pathway and to compare the gene profiles between human muscle biopsies and cultured muscle cells from the same donors. To identify susceptibility genes for obesity, we have performed a gene expression profiling experiment using freshly isolated adipocytes from 20 lean and 19 obese Pima Indians. We are currently analyzing the data, and will pursue potential susceptibility genes for obesity based on functions and chromosomal locations with linkage to obesity measures.