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Methylation analysis of the gene ADRB2

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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 disease. We have recent preliminary data showing that in human airway smooth muscle (HASM) cells 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. .=. 0-0 'm :3 M-0 W-0 .4) v+3 'Z-_ U).2 Gam. =:r a:' pro. a-- --, >,Q -0a any tom/!
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