MOLECULAR TARGETS OF PERINATAL ENDOCRINE DISRUPTION

围产期内分泌干扰的分子目标

• 批准号: 2881611
• 负责人: WILLIAM J HENDRY
• 金额: $ 10.72万
• 依托单位: WICHITA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-15 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2881611
• 关键词: diethylstilbestrol; drug adverse effect; female reproductive system disorder; gene expression; hamsters; hormone regulation /control mechanism; hormone related neoplasm /cancer; molecular cloning; newborn animals; reproductive development; uterus; uterus preneoplastic state

From our data showing that neonatal treatment with the classic endocrine-disrupting agent diethylstilbestrol (DES) acts to alter estrogen responsiveness in the adult hamster uterus by a direct mechanism, we propose two related working hypotheses are: 1) The altered estrogen responsiveness that occurs in the adult hamster uterus is the result of neonatal DES-induced imbalances in the expression of key regulatory genes during early development; and 2) Stromal-epithelial interactions make it likely that DES- altered patterns of early gene expression will differ between the two tissue compartments. Based on these hypotheses, we will pursue the Specific Aim to: Identify the immediate cellular and molecular targets of the neonatal DES insult that cause inappropriate estrogen responses in the adult hamster uterus. Fortunately, improved versions of powerful new genetic screening and cloning strategies are now available in kit form and require very little total cellular RNA as starting material. Also, determining the cellular site of the immediate DES-induced alterations will be facilitated by our ability to cleanly separate the simple luminal epithelium from the relatively undifferentiated stromal compartment that is present in neonatal uteri. This screening effort should 1) generate a succession of newly identified genetic elements, several of which must be involved in important biological functions (estrogen-responsive proliferation/apoptosis/neoplasia), and 2) be followed by an ordered determination of each element s molecular structure and functional dynamics at the genomic, RNA, and protein level. The spectrum of research opportunities defined by each such process would encompass efforts at many levels including those appropriate for several of our independent faculty members, for post-doctoral research associates, or for our students at either the graduate or undergraduate level. Thus this project is particularly relevant to the stated purpose of the AREA Award Program. In addition to providing important mechanistic information about the controversial topic of perinatal endocrine disruption, this project will surely benefit our larger long-term objectives to: 1) Delineate the basic mechanisms whereby estrogen regulates normal uterine growth and morphogenesis, and 2) Identify mechanistic alterations that cause degeneration of the normal growth process and thereby leads to the unregulated neoplastic state. These are biomedically important because: 1) Successful conception and gestation demands normal uterine form and function, and 2) Estrogen-dependent uterine neoplasms are responsible for considerable morbidity and mortality in contemporary American society.

根据我们的数据显示，使用经典内分泌干扰剂己烯雌酚 (DES) 进行新生儿治疗可通过直接机制改变成年仓鼠子宫中的雌激素反应性，我们提出两个相关的工作假设：1) 成年仓鼠子宫中发生的雌激素反应性改变是新生儿 DES 诱导的早期发育过程中关键调控基因表达失衡的结果； 2) 基质-上皮相互作用使得 DES 改变的早期基因表达模式可能在两个组织区室之间有所不同。 基于这些假设，我们将追求的具体目标是：确定导致成年仓鼠子宫中不适当雌激素反应的新生儿 DES 损伤的直接细胞和分子靶点。幸运的是，强大的新基因筛选和克隆策略的改进版本现已以试剂盒形式提供，并且只需要很少的总细胞 RNA 作为起始材料。 此外，我们能够将简单的管腔上皮与新生儿子宫中存在的相对未分化的基质区室干净地分离，从而有助于确定 DES 诱导的直接改变的细胞位点。 这种筛选工作应该1）产生一系列新鉴定的遗传元件，其中一些必须涉及重要的生物功能（雌激素反应性增殖/细胞凋亡/肿瘤），2）随后在基因组、RNA和蛋白质水平上有序确定每个元件的分子结构和功能动态。 每个这样的过程定义的研究机会范围将涵盖许多层面的努力，包括适合我们的几位独立教员、博士后研究员或我们的研究生或本科生的努力。 因此，该项目与 AREA 奖励计划的既定目的特别相关。 除了提供有关围产期内分泌干扰这一有争议话题的重要机制信息外，该项目肯定有利于我们更大的长期目标：1）描述雌激素调节正常子宫生长和形态发生的基本机制，2）识别导致正常生长过程退化并从而导致不受调节的肿瘤状态的机制改变。 这些在生物医学上很重要，因为：1）成功的受孕和妊娠需要正常的子宫形态和功能，2）雌激素依赖性子宫肿瘤是当代美国社会相当大的发病率和死亡率的原因。