PEPTIDE GROWTH FACTORS IN POLYCYSTIC OVARIAN DISEASE

多囊卵巢疾病中的肽生长因子

基本信息

批准号：
3426740
负责人：
JOHN YEH
金额：
$ 3.41万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-08-01 至 1993-07-31
项目状态：
已结题

项目摘要

Polycystic ovarian disease (PCOD) is a common gynecologic problem in which patients suffer from hyperandrogenism, hirsutism and infertility. The role of peptide growth factors as autocrine or paracrine contributors to this disease is unclear. E. Anderson (see collaboration letter) and others have developed a rat model for polycystic ovary in which the in vitro and in vivo effects of hyperandrogenism on granulosa cells can be tested experimentally. The goal of this proposal is to study, by using the rat model, the potential roles of transforming growth factor-alpha (TGF-a), epidermal growth factor (EGF) (the biochemical and functional analog of TGF-a), and their common receptor, the EGF receptor, in the pathophysiology of PCOD. Specific aims are: I. To study TGF-a and granulosa function in cells subjected to hyperandrogenic environments. Preliminary results indicate that normal granulosa cells contain TGF-a mRNA and respond to exogenously added TGF-a. These experiments are planned: (1) In vitro, rat granulosa cells cultured with androgens and TGF-a or EGF will be studied for changes in TGF-a mRNA production, in estrogen and progesterone production, in levels of mRNA of the enzymes of steroidogenesis, in the kinetics of DNA synthesis and in cellular morphology. (2) In vivo, the change in TGF-a gene expression during polycystic ovary formation in rats injected with dehydroepiandrosterone will be analyzed by in situ hybridization and immunohistochemistry. II. To understand TGF-a signal transduction in hyperandrogenic granulosa cells. Normal granulosa cells contain receptors for TGF-a. The effects of hyperandrogenism on TGF-a signal transduction are unknown. These experiments are planned: (1) Analysis of changes in EGF receptors in hyperandrogenism in terms of change in receptor binding affinity, in level of EGF receptor mRNA production, in cellular localization and in subcellular (vesicular/cytoskeletal) localization. (2) Study of blockage of TGF-a signal transduction in hyperandrogenism. TGF-a signal transduction will be blocked with anti-TGF-a antibodies and with genistein (a tyrosine kinase inhibitor) and the cells will be analyzed for these changes: TGF-a mRNA expression; c-fos and c-myc proto-oncogene mRNA expression; steroid production; and morphology. III. To investigate the regulation of TGF-a gene expression in hyperandrogenic granulosa cells. TGF-a gene expression is inducible with FSH and with estradiol. The rat TGF-a gene (6 exons, 85 kilobases in length) has been mapped and the promoter region has recently been sequenced. It is planned that the FSH and estradiol responsive sites of the TGF-a gene be identified and elucidated. After that, the regulation of TGF-a gene expression in hyperandrogenism will be compared to normal.

多囊卵巢疾病（PCOD）是一个常见的妇科问题，其中患者患有高狂力，多毛和不育。角色肽生长因子是自分泌或旁分泌因素疾病尚不清楚。 E.安德森（请参阅协作信）等开发了一种大鼠模型的多囊卵巢模型，其中体外和中可以测试高狂力主义对颗粒细胞的体内影响实验。该建议的目的是使用老鼠研究模型，转化生长因子-Alpha（TGF-A）的潜在作用，表皮生长因子（EGF）（生化和功能类似物 TGF-A）及其公共受体EGF受体在病理生理学中 PCOD。具体目的是： I.在受到的细胞中研究TGF-A和颗粒功能高雄激素环境。初步结果表明正常颗粒细胞含有TGF-A mRNA，并响应外源添加的TGF-A。这些实验是计划的：（1）在体外，大鼠颗粒细胞培养将研究使用雄激素和TGF-A或EGF进行TGF-A mRNA的更改在雌激素和孕酮产生中生产mRNA的水平类固醇生成的酶，在DNA合成的动力学和细胞形态。（2）在体内，TGF-A基因表达的变化在注射大鼠的多囊卵巢形成期间将通过原位杂交和免疫组织化学。 ii。了解高雄激素颗粒中的TGF-A信号转导细胞。正常的颗粒细胞包含用于TGF-A的受体。效果 TGF-A信号转导的过度狂力尚不清楚。这些计划实验：（1）分析EGF受体的变化在受体结合亲和力的变化方面，高雄激素在水平上 EGF受体mRNA的产生，细胞定位和亚细胞（囊泡/细胞骨架）定位。（2）堵塞研究高雄激素中TGF-A信号转导的。 TGF-A信号转导将用抗TGF-A抗体阻塞，并用染料木黄酮阻塞（酪氨酸激酶抑制剂），将分析细胞变化：TGF-A mRNA表达； C-FOS和C-MYC原始癌基因mRNA 表达;类固醇产生；和形态。 iii。研究TGF-A基因表达中的调节高雌激素颗粒细胞。 TGF-A基因表达可诱导 FSH和雌二醇。大鼠TGF-A基因（6个外显子，85千碱基长度）已被映射，启动子区域最近已测序。据计划，FSH和雌二醇的响应位置鉴定和阐明TGF-A基因。之后，调节高雄激素中的TGF-A基因表达将与正常情况进行比较。