Genome wide DNA methylation and Hydroxymethylation of F1 mouse lung prenatally exposed to house dust mite. Genome wide DNA methylation and Hydroxymethylation of F1 mouse lung prenatally exposed to house dust mite

We used a mouse model of allergic lung disease to examine the effects of pre- and perinatal house dust mite (HDM) allergen exposure on offspring lung phenotypic and transcriptional outcomes. We showed that maternal HDM exposure (F0) acts synergistically with adult HDM exposure, leading to enhanced airway hyperresponsiveness (AHR) and lung inflammation when compared to mice exposed solely in adulthood. To examine the role of epigenetic inheritance of asthma susceptibility induced by maternal HDM exposure, we utilized a genome-wide MeDIP-seq and hMeDIP-seq analysis to identify genes differentially methylated (DMG) and hydroxymethylated (DHG), and their association with the enhanced AHR. In addition, we validated the relationship between DNA methylation and mRNA expression of the DMGs and DHGs in the male progenies. Overall design: We compared the lung DNA methylation and hydroxymethylation patterns from F1 progenies from dams exposed to saline or HDM (F0-Saline_F1-HDM or F0-HDM_F1-HDM) when compared to their counterparts who did not received HDM challenges in their adulthood (F0-Saline_F1-Saline or F0-HDM_F1-Saline). Genomic DNA from each treatment group (pooled from mice of 3 individual dams) was subject for MeDIP-seq and hMeDIP-seq. Validation of airway responsiveness, cell counts and relative gene expression were performed on six mice per treatment group.

我们采用过敏性肺病小鼠模型，探究产前与围产期屋尘螨（house dust mite, HDM）过敏原暴露对子代肺表型及转录组特征的影响。研究发现，母代（F0）HDM暴露与成年期HDM暴露存在协同效应：相较于仅在成年期暴露于HDM的小鼠，子代的气道高反应性（airway hyperresponsiveness, AHR）与肺部炎症水平均显著升高。为探究母代HDM暴露诱导的哮喘易感性状的表观遗传传递机制，我们采用全基因组MeDIP-seq与hMeDIP-seq分析，筛选差异甲基化基因（differentially methylated gene, DMG）与差异羟甲基化基因（differentially hydroxymethylated gene, DHG），并分析其与气道高反应性增强的关联。此外，我们在雄性子代中验证了差异甲基化基因与差异羟甲基化基因的DNA甲基化水平与其mRNA表达量之间的关联。实验整体设计如下：我们对F1子代的肺部DNA甲基化与羟甲基化模式进行分组比较，分组依据为母代暴露生理盐水或HDM，以及子代成年期是否接受HDM刺激：具体包括母代生理盐水暴露+子代成年HDM暴露组（F0-Saline_F1-HDM）、母代HDM暴露+子代成年HDM暴露组（F0-HDM_F1-HDM），以及对应的对照组：母代生理盐水暴露+子代成年未接受HDM刺激组（F0-Saline_F1-Saline）、母代HDM暴露+子代成年未接受HDM刺激组（F0-HDM_F1-Saline）。每个处理组的基因组DNA（由3只母代小鼠的子代混合提取制备）均进行MeDIP-seq与hMeDIP-seq测序。我们对每个处理组的6只小鼠开展了气道反应性、细胞计数与相关基因相对表达量的验证实验。