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子，胰岛素抵抗和肥胖的易感基因。为此，我们研究了骨骼肌基因表达谱在非糖尿病的纯种皮马印第安人使用1）人寡核苷酸微阵列含有6800个基因和35000表达序列标签（EST）和2）差异显示PCR，然后通过真实的时间RT-PCR。这项工作已发表在Diabetologia，DOI 10.1007/s 00125 -002-0905-7。通过检查1）它们在相关代谢途径中的功能，如IRS-1（Diabetes 51，Supplement 2：A266）和2）它们在Pima群体中的全基因组扫描中相对于与胰岛素作用和糖尿病的连锁峰的染色体位置，如ORP 150（Diabetes 51：1618-1621），评价差异表达基因/EST在胰岛素抗性中的潜在作用。 我们还将使用原代人骨骼肌培养系统来鉴定胰岛素信号通路的潜在新成员，并比较来自相同供体的人肌肉活检和培养的肌肉细胞之间的基因谱。 为了确定肥胖的易感基因，我们进行了基因表达谱实验，使用新鲜分离的脂肪细胞从20瘦和19肥胖皮马印第安人。我们目前正在分析这些数据，并将根据与肥胖相关的功能和染色体位置来寻找肥胖的潜在易感基因。

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.