Trophoblasts and Inflammation: An Epigenetic Approach

描述(申请人提供)：事实证明，人类基因组是一个非常忙碌和有趣的地方。基因组不是基因转录的被动模板，而是细胞功能的组成部分，对外部环境信号高度敏感，并能够通过阅读和自我调节来对这些信号做出反应。我们假设，许多以慢性、低级别炎症为特征的人类疾病的出现，是因为环境诱导的表观遗传变化损害了基因组读取和调节自身的能力。虽然许多关于表观遗传对基因转录的影响的研究都集中在蛋白质编码基因上，但我们现在知道，表观遗传改变和RNA表达都发生在整个基因组中，而不仅仅是在注释良好的基因内或附近。因此，当我们的团队在体外人类绒毛膜癌JEG-3细胞炎症模型中发现与基因表达变化相关的甲基化变化时，我们并不惊讶地发现，甲基化发生在距离差异表达基因相当远的基因组区域。我们急于了解这些差异甲基化区域的情况，因为它们可能会告诉我们很多关于滋养层细胞如何在全基因组基础上对炎症做出反应的信息。然而，在我们做其他任何事情之前，我们需要在一个与病理相关的细胞中复制这些数据，即来自胎膜的绒毛膜滋养层细胞。在我们这样做的同时，我们也有机会对滋养细胞转录组有更丰富和更好的了解。我们建议使用最先进的测序技术对足月绒毛滋养细胞进行基因组范围的甲基化分析和基因组范围的转录组分析。我们将把RNA转录物映射到基因组，并将转录物和转录物水平与炎症引起的甲基化变化联系起来。两年后，我们将拥有一个非常有用的数据集，从中我们可以组织面向患者的研究和体外研究，重点是妊娠期间的炎症和功能基因组学。

DESCRIPTION (provided by applicant): The human genome, it turns out, is a very busy and interesting place. Rather than serving as a passive template for gene transcription, the genome is an integral part of cell function, highly sensitive to external environmental cues and capable o responding to those cues by reading and regulating itself. We have hypothesized that many human diseases characterized by chronic, low-grade inflammation, emerge because the capacity of the genome to read and regulate itself is compromised by environmentally-induced epigenetic alterations. While multiple investigations into epigenetic effects on gene transcriptio have focused on protein-coding genes, we now know that both epigenetic alterations and RNA expression occur throughout the genome, not simply within or adjacent to well-annotated genes. Thus, when our group found methylation changes associated with gene expression changes in an in vitro model of inflammation in human choriocarcinoma Jeg-3 cells, we were not surprised to find that that methylation occurred in genomic regions quite distant from the differentially-expressed genes. We are anxious to know what's in those differentially-methylated regions, as they are likely to tell us a great deal about how trophoblasts respond to inflammation on a genome-wide basis. Before we do anything else, however, we need to reproduce these data in a pathologically-relevant cell, i.e., term chorionic trophoblasts from fetal membranes. While we're doing that, we also have the opportunity to geta richer and better understanding of the trophoblast transcriptome. We propose to perform genome-wide methylation analysis and genome-wide transcriptome analysis on term chorionic trophoblasts using state-of-the art sequencing techniques. We will map RNA transcripts to the genome and correlate transcripts and transcript levels with methylation changes that occur as a result of inflammation. After 2 years, we will have an extraordinarily useful data set from which we can organize both patient-oriented and in vitro studies focusing on inflammation and functional genomics in pregnancy.