Epigenetic regulation of decidual inflammation

描述（由申请人提供）：由于我们目前对潜在的致病机制缺乏了解，预防早产和其他人类妊娠并发症的策略的发展受到了极大的阻碍。特别是，关于蜕膜的内在炎症特征知之甚少，蜕膜是包围胎儿和胎盘的特殊子宫间质组织。这一建议是基于我的实验室最近对怀孕小鼠子宫中控制蜕膜炎症和白细胞运输的分子途径的研究。我们发现，子宫内膜基质细胞（ESCs）向蜕膜基质细胞（dsc）的分化需要通过H3K27me3抑制组蛋白标记的启动子积累来抑制炎性趋化因子基因的表观遗传沉默。这种新的发育程序显著降低了蜕膜表现炎症反应和积累活化T细胞的潜力。在未发表的数据中，我们发现DSC分化还需要沉默编码多功能趋化因子Cxcl12的Cxcl12和编码巨噬细胞生长因子CSF-1的Csf1。总之，这些发现提出了类似的“蜕膜基因沉默”程序是否在人类dsc中活跃的问题，并表明该程序的失调可能是各种胎盘/蜕膜病理的基础。该提案分为三个具体目标。Aim I试图更深入地了解蜕膜基因沉默的广度、分子特征和功能意义，再次使用小鼠作为模型生物。这些实验将为评估人类妊娠中的蜕膜基因沉默，以及考虑其对人类妊娠并发症的潜在意义奠定基础。Aim II是该提案的主要部分，它采用新鲜的妊娠早期蜕膜标本和许多生化技术来确定蜕膜基因沉默是否是人类妊娠的一个特征。Aim III开发了一种新的组蛋白标记免疫染色/DNA荧光原位杂交（DNA- fish）方案，用于检测单细胞水平上的单个基因沉默程序，从而最终检测到该程序的潜在失调

DESCRIPTION (provided by applicant): The development of strategies to prevent preterm birth and other complications of human pregnancy has been greatly hindered by our current lack of understanding of underlying pathogenic mechanisms. In particular, little is known about the intrinsic inflammatory characteristics of the decidua, the specialized uterine stromal tissue that surrounds the fetus and placenta. This proposal is based upon recent work from my laboratory on the molecular pathways that control decidual inflammation and leukocyte trafficking in the pregnant mouse uterus. We have found that the differentiation of endometrial stromal cells (ESCs) into decidual stromal cells (DSCs) entails the epigenetic silencing of select inflammatory chemokine genes through their promoter accrual of the H3K27me3 repressive histone mark. This novel developmental program dramatically reduces the potential of the decidua to manifest an inflammatory response and accumulate activated T cells. In unpublished data, we have found that DSC differentiation also entails the silencing of Cxcl12 encoding the multifunctional chemokine CXCL12, and Csf1 encoding the macrophage growth factor CSF-1. Together, these findings raise the question of whether an analogous "decidual gene silencing" program is active in human DSCs, and suggest that dysregulation of this program may underlie a variety of placental/decidual pathologies. The proposal is divided into three Specific Aims. Aim I seeks to gain greater insight into the breadth, molecular characteristics, and functional significance of decidual gene silencing, again using mice as a model organism. These experiments will establish a foundation for evaluating decidual gene silencing in human pregnancy, and for considering its potential significance with regards to human pregnancy complications. Aim II, which constitutes the bulk of the proposal, employs fresh first trimester decidual specimens and a number of biochemical techniques to determine whether decidual gene silencing is a feature of human pregnancy. Aim III develops a novel histone mark immunostaining/DNA fluorescence in situ hybridization (DNA-FISH) protocol for detecting the decidual gene silencing program at the single cell level so that potential dysregulation of this program can ultimately be detected in archived placental/decidual pathological specimens.