UTERINE DISRUPTION: DNA METHYLATION & EPITHELIA-STROMA

描述(由申请人提供)：长期目标是：1)确定雌激素调节生殖道器官细胞增殖和组织形态形成的正常模式的基本机制；以及2)确定正常雌激素反应中断与特定细胞事件和分子因素变化之间的因果联系。PI已经确定，用已建立的内分泌干扰剂己烯雌酚(DES)治疗新生儿可以直接和永久性地改变发育中的仓鼠子宫(发育期)，使成年仓鼠子宫对天然雌激素雌二醇的刺激(促进期)做出异常反应。符合这一现象并响应计划公告的胎儿/发育重点的主要工作假说(WH)是：1‘WH--在早期发育期间DNA甲基化的变化是新生儿DES暴露永久扰乱子宫形态发生和雌激素反应的机制的一部分。第二个工作假说结合了有关雌激素反应器官发育和功能调节机制的新信息：2‘WH-上皮-间质(间质)相互作用参与子宫发育和功能，使得新生儿甲基化的程度或模式在两个组织区段之间可能不同。下面的一组具体目标(SA)代表了一种创新但可行的策略来检验这些相互关联的假设。SA#1-筛选在新生儿DES诱导的子宫破裂的初始阶段发生的DNA甲基化的组织特异性差异的证据。SA#2-确定来自筛查的产物是否与已知基因或先前未识别的(新的)基因相关联，这些基因在对照和新生儿DES干扰的仓鼠子宫中差异表达。SA#3-在组织特异性水平上，评估新生儿DES诱导的DNA甲基化变化与随后仓鼠子宫内基因表达变化之间的关系。对这种细胞间和基因组动力学的更全面的了解将有助于产生新的战略：1)更好地评估疾病进展的阶段/等级，以及2)更好地选择和提供治疗剂。它还应该为“内分泌干扰”这一话题带来重要的新见解。

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'.