UTERINE DISRUPTION: DNA METHYLATION & EPITHELIA-STROMA

描述（由申请方提供）：长期目标是：1）确定雌激素调节生殖道器官中细胞增殖和组织形态发生正常模式的基本机制; 2）确定正常雌激素反应中断与特定细胞事件和分子因子改变之间的因果关系。PI已经确定，使用已确定的内分泌干扰剂己烯雌酚（DES）对新生儿进行治疗，直接并永久性地改变了发育中的仓鼠子宫（起始阶段），从而使成年仓鼠子宫对天然雌激素雌二醇的刺激（促进阶段）产生异常反应。与该现象一致并且响应于该计划公告的胎儿/发育焦点的主要工作假设（WH）是：1'WH-早期发育期间DNA甲基化的改变是新生儿DES暴露永久破坏子宫形态发生和雌激素反应性的机制的一部分。第二个工作假设，它结合了关于调节雌激素反应器官的发育和功能的机制的新信息：2'WH-子宫发育和功能中上皮-基质（间充质）相互作用的参与使得新生儿DES改变的甲基化的程度或模式可能在两个组织区室之间不同。以下一组具体目标是一种创新但可行的战略，以检验这些相关的假设。SA#1-筛查新生儿DES诱导的子宫破裂起始阶段发生的DNA甲基化组织特异性差异的证据。SA#2-确定筛选产物是否与已知基因或先前未识别（新）基因相关，这些基因在对照与经DES破坏的仓鼠子宫中差异表达。SA#3-在组织特异性水平上，评价新生DES诱导的DNA甲基化改变与仓鼠子宫内基因表达随后变化之间的关系。对这种细胞间和基因组动力学的更全面理解将有助于产生新的策略：1）更好地评估疾病进展的阶段/等级，2）更好地选择和提供治疗药物。它还应该对“内分泌干扰”这一主题产生重要的新见解。

DESCRIPTION (provided by applicant): The long-term objectives are to: 1) determine the basic mechanisms whereby estrogen regulates normal patterns of cell proliferation and tissue morphogenesis in reproductive tract organs; and 2) identify causal links between disruption of normal estrogen responses and alterations in specific cellular events and molecular factors. The PI has determined that neonatal treatment with the established endocrine-disrupting agent, diethylstilbestrol (DES), directly and permanently alters the developing hamster uterus (initiation phase) so that the adult hamster uterus responds abnormally to stimulation (promotion phase) with the natural estrogen, estradiol. A primary working hypothesis (WH) that is consistent with that phenomenon and is responsive to the fetal/developmental focus of the Program Announcement is: 1' WH-Alterations in DNA methylation during early development are part of the mechanism by which neonatal DES exposure permanently disrupts uterine morphogenesis and estrogen responsiveness. A secondary working hypothesis that incorporates new information about the mechanisms that regulate development and function of estrogen-responsive organs is: 2' WH-The involvement of epithelial-stromal (mesenchymal) interactions in uterine development and function makes it likely that the extent or pattern of neonatal DES-altered methylation will differ between the two tissue compartments. The following set of Specific Aims (SA) represents an innovative but feasible strategy to test those linked hypotheses. SA#1-Screen for evidence of tissue specific differences in DNA methylation that occur during the initiation phase of neonatal DES-induced uterine disruption. SA#2-Determine if products from the screen are linked either to known genes or to previously unidentified (novel) genes that are differentially expressed in the control vs. neonatally DES-disrupted hamster uterus. SA#3-At the tissue-specific level, evaluate the relationship between neonatal DES-induced alterations in DNA methylation and subsequent changes in gene expression within the hamster uterus. A more complete understanding of such intercellular and genomic dynamics will help generate new strategies to: 1) better evaluate the stage/grade of disease progression, and 2) better choose and deliver therapeutic agents. It should also yield important new insight into the topic of 'endocrine disruption'.