17 alpha-Hydroxylase Expression in Human Ovarian Cells

17 α-羟化酶在人卵巢细胞中的表达

• 批准号: 7766295
• 负责人: Jan M McAllister
• 金额: $ 28.64万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7766295
• 关键词: Abbreviations; Adenoviruses; Affect; Anabolism; Androgens; Area; CYP11A1 gene; Cell physiology; Cells; Cholesterol; Cytochrome P450; Data; Defect; Development; Dominant-Negative Mutation; Dose; Event; Follicular Fluid; Funding; Gene Expression; Genetic Transcription; Goals; Human; Infection; Laboratories; Lead; Lesion; Literature; MAP2K1 gene; MAP3K1 gene; MAPK14 gene; MEKKs; Messenger RNA; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Mitogens; Mixed Function Oxygenases; Molecular; Motion; Ovarian; Ovary; Pathway interactions; Phenotype; Phosphotransferases; Plasma; Polycystic Ovary Syndrome; Production; Regulation; Reporting; Research Design; Resistance; Side; Signal Pathway; Signal Transduction; Steroid 17-alpha-monooxygenase; Stream; System; Testing; Theca folliculi structure; Transcriptional Regulation; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Woman; base; dehydroepiandrosterone; design; insight; mRNA Stability; novel; promoter; receptor; research study; response; theca cell; tool

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.

提出的实验旨在阐明雄激素基础的调节机制 来自正常骑自行车妇女和多囊卵巢妇女的卵巢的theca国内细胞中的生物合成 综合征（PCOS）。在过去的资金期间，我们获得了令人兴奋的数据，表明增加了 PCOS theca细胞中的雄激素产生和增强P450 17A-羟化酶（CYP17）基因表达 MEK1/ERK途径的激活状态的综合减少以及刺激的结果 MKK3途径的激活状态。我们已经观察到与正常theca细胞感染 腺病毒表达显性负MEK1并组成性活性MKK3概括了PCOS theca 细胞表型。现在，我们拥有充分探索其他组件的贡献的工具 MAPK信号通路。在此提案中，我们将检验以下假设，即增加卵巢雄激素 生产是PCOS THECA中有丝分裂原活化蛋白激酶（MAPK）信号改变的结果 细胞。 AIM 1研究旨在研究上游信号传导分量的失调是否失调 ERK和p38信号通路（s）（即RAF-1，MEKK1，MEKK3）直接影响向下流 CYP17和CYP11A1基因的范围，每个同源途径的信号传导成分 在PCOS THECA细胞中，表达和雄激素生物合成被上调。我们还将检查是否 信号传导的补偿性变化中，来自一个途径结果的信号传导分量失调 通过平行路径（S）。在AIM 2中，我们将研究ERK和P38的改变程度 信号导致PCOS中CYP17和CYP11A1基因转录和mRNA稳定性增加 theca细胞。 在整个文献中，有数据支持肿瘤坏死因子A（TNF）抑制了疾病 CYP17基因表达和雄激素生物合成。但是，这些数据与 观察到PCOS妇女的血浆循环升高和TNF的卵泡流体水平升高。在AIM 3中， 我们将研究PCOS theca细胞中MAPK信号中缺陷的程度影响TNF依赖性 调节雄激素生物合成。这些研究的结果将提供有关有关的新见解 增加卵巢雄激素产生的原因和分子基础，并导致新的发展 PCOS女性治疗的靶标。

项目成果

Single-Cell RNA-Seq Identifies Pathways and Genes Contributing to the Hyperandrogenemia Associated with Polycystic Ovary Syndrome.

• DOI: 10.3390/ijms241310611
• 发表时间: 2023-06-25
• 期刊: International journal of molecular sciences
• 影响因子: 5.6

The Cebpd (C/EBPdelta) gene is induced by luteinizing hormones in ovarian theca and interstitial cells but is not essential for mouse ovary function.

• DOI: 10.1371/journal.pone.0001334
• 发表时间: 2007-12-19
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Huang AM;Rudelius M;Sharan S;McAllister JM;Raffeld M;Christenson LK;Sterneck E
• 通讯作者: Sterneck E