Methylation analysis of the gene ADRB2

基因表达的表观遗传调控是一个与人类健康相关的新兴概念。该提案的目的是研究与阻塞性肺相关的细胞中ADRI32表达的表观遗传学控制 .=. �‘m 暴露在a-激动剂下，13-激动剂受体ADRB2基因的启动子和编码区的DNA甲基化程度增加。基于DNA甲基化阻断或降低基因表达的经典功能，我们假设HASM细胞长期暴露于13-激动剂会增加ADRB2的甲基化，导致ADRB2表达下降。ADR132的这种减少可能是导致哮喘和COPD患者使用13种激动剂治疗的快速反应的主要机制。在这项拟议的初步研究中，我们将通过以下方式验证我们的假设：1)对ADRI32进行甲基化特异性测序，以定位和定量从暴露于13-激动剂的HASM细胞中分离的DNA中甲基化的CpG位点；2)评估ADR2甲基化对基因表达的影响。 V+3 �：3 W-0 =：R 答：‘ �--、 任何 0-0 疾病。我们最近的初步数据显示，在人类呼吸道平滑肌(HASM)细胞中 U).2 M-0 .4) `Z-_ 阿甘。 职业选手。 -0A� &gt；，Q 汤姆/！ A--

Epigenetic regulation of gene expression is an emerging concept relevant to human health. This proposal's objective is to investigate the epigenetic control of ADRI32 expression in cells relevant to obstructive lung .=. �'m exposed to a a-agonist, the degree of DNA methylation is increased in the promoter and coding region of the gene for the 13-agonist receptor, ADRB2. Based on the classical function that DNA methylation blocks or decreases gene expression, we hypothesize that long term exposure of HASM cells to 13-agonist increases methylation of ADRB2 and resulting in decreased expression of ADRB2. This decrease in ADR132 could be a major mechanism contributing to tachyphylaxis to 13-agonist treatment in asthma and COPD. In this proposed pilot study, we will test our hypothesis by: 1) Performing methylation specific sequencing on ADRI32 to localize and quantitate methylated CpG sites in DNA isolated from HASM cells exposed to 13-agonists; 2) Assessing the effects of ADR2 methylation on gene expression. v+3 �:3 W-0 =:r a:' �--, any 0-0 disease. We have recent preliminary data showing that in human airway smooth muscle (HASM) cells U).2 M-0 .4) `Z-_ Gam. pro. -0a� >,Q tom/! a--