17 alpha-Hydroxylase Expression in Human Ovarian Cells

这些实验旨在阐明雄激素的调节机制 正常周期妇女和多囊卵巢妇女卵巢内膜细胞生物合成 综合征（PCOS）。在过去的资助期间，我们获得了令人兴奋的数据，表明增加了 PCOS卵泡膜细胞中雄激素的产生和P450 17 α-羟化酶（CYP 17）基因的表达增强 这是由于MEK 1/ERK通路的激活状态的降低和细胞内信号转导通路的刺激的联合作用。 MKK 3通路的激活状态。我们观察到正常卵泡膜细胞感染 表达显性阴性MEK 1和组成型活性MKK 3的腺病毒重现PCOS卵泡膜 细胞表型我们现在有工具来充分探讨其他组成部分的贡献， MAPK信号通路。在这个提议中，我们将检验卵巢雄激素增加的假说， 在PCOS卵泡膜中， 细胞目的1研究上游信号成分的失调是否 ERK和p38信号传导途径（即，Raf-1、MEKK 1、MEKK 3）直接影响下游 在某种程度上，CYP 17和CYP 11 A1基因 表达和雄激素生物合成在PCOS卵泡膜细胞中上调。我们还将研究 来自一种途径的信号传导组分的失调导致信号传导的代偿性变化 通过平行路径（S）。在目标2中，我们将研究ERK和p38的改变在多大程度上影响了 多囊卵巢综合征中CYP 17和CYP 11 A1基因转录和mRNA稳定性增加 泡膜细胞 在整个文献中，有数据支持肿瘤坏死因子a（TNF）对鞘内 CYP 17基因表达和雄激素生物合成。然而，这些数据与 观察到PCOS妇女循环血浆和卵泡液中TNF水平升高。在目标3中， 我们将研究PCOS卵泡膜细胞中MAPK信号的缺陷在多大程度上影响TNF依赖性 雄激素生物合成的调节。这些研究的结果将提供关于 卵巢雄激素产生增加的原因和分子基础，并导致新的 治疗PCOS女性的目标。

The proposed experiments are designed to elucidate the regulatory mechanisms underlying androgen biosynthesis in theca interna cells from ovaries of normal cycling women and women with polycystic ovary syndrome (PCOS). During the past funding period, we obtained exciting data demonstrating that increased androgen production and augmented P450 17a-hydroxylase (CYP17) gene expression in PCOS theca cells results from a combined reduction in the activation state of the MEK1/ERK pathway and a stimulation in the activation state of the MKK3 pathway. We have observed that infection of normal theca cells with adenovirus expressing dominant negative MEK1 and constitutively active MKK3 recapitulates a PCOS theca cell phenotype. We now have the tools to fully explore the contributions of the other components of the MAPK signaling pathways. In this proposal we will test the hypothesis that increased ovarian androgen production is a consequence of altered mitogen activated protein kinase (MAPK) signaling in PCOS theca cells. Aim 1 studies are designed to investigate whether dysregulation of upstream signaling component(s) of the ERK and p38 signaling pathway(s) (i.e., Raf-1, MEKK1, MEKK3) directly affect the down stream signaling components of each homologous pathway to the extent that CYP17 and CYP11A1 gene expression, and androgen biosynthesis are up-regulated in PCOS theca cells. We will also examine whether dysregulation of signaling components from one pathway result(s) in compensatory changes in signaling through parallel pathway(s). In Aim 2 we will investigate the extent to which alterations in ERK and p38 signaling contribute to increased CYP17 and CYP11A1 gene transcription and mRNA stability in PCOS theca cells. Throughout the literature there are data to support that tumor necrosis factor a (TNF) inhibitsthecal CYP17 gene expression and androgen biosynthesis. However, these data are contradictory with the observation that PCOS women have elevated circulating plasma and follicular fluid levels of TNF. In Aim 3, we will examine the extent to which defects in MAPK signaling in PCOS theca cells affects TNF-dependent regulation of androgen biosynthesis. Results of these studies will provide both new insights regarding the cause and molecular basis for increased ovarian androgen production, and lead to the development of new targets for treatment of women with PCOS.