Functional Significance of the HIF1 Transcriptome in Granulosa Cells

颗粒细胞中 HIF1 转录组的功能意义

基本信息

批准号：
8113398
负责人：
Mary E Hunzicker-Dunn
金额：
$ 29.29万
依托单位：
WASHINGTON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-01 至 2015-07-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8113398
关键词：
1-Phosphatidylinositol 3-Kinase Basement membrane Biological Assay Blood capillaries Cell Culture Techniques Cell Extracts Cell Hypoxia Cell Maturation Cell physiology Cells Chorionic Gonadotropin Contraceptive Agents Cyclic AMP-Dependent Protein Kinases Development Dominant-Negative Mutation Enzymes Female Fertility Follicle Stimulating Hormone G-Protein-Coupled Receptors Gene Expression Microarray Analysis Gene Expression Profile Gene Targeting Genes Graafian Follicles Growing Follicle Growth Growth Factor Hormones Housing Hypoxia Hypoxia Inducible Factor Infertility Luteinizing Hormone Maps Molecular Profiling Mus Nutrient Oocytes Ovarian Follicle Oxygen Pathway interactions Pharmaceutical Preparations Phase Phenotype Phospho-Specific Antibodies Pituitary Hormones Post-Translational Protein Processing Pregnancy loss Process Proteins Regulation Role Signal Transduction Site Staging Steroids Surface Testing Transcription Coactivator Translating Translations Vascular Endothelial Growth Factor Receptor Vascular Endothelial Growth Factors Vascularization Western Blotting autocrine base cancer cell capillary chromatin immunoprecipitation cohort effective therapy genome sequencing granulosa cell growth hormone regulating factor hormone regulation hypothalamic pituitary axis hypoxia inducible factor 1 inhibin inhibitor/antagonist mRNA Expression mTOR protein novel strategies oocyte maturation public health relevance receptor recombinase research study response small hairpin RNA theca cell uterine receptivity

项目摘要

DESCRIPTION (provided by applicant): Fertility in females requires controlled maturation of the oocyte, supporting granulosa cells (GCs), and theca cells that comprise the ovarian follicle. Nutrients and hormones to support follicle maturation come from capillaries in the theca layer; vascularization terminates at the basement membrane. Thus, as follicles grow, GCs are exposed to increasingly hypoxic conditions. Follicle growth is a dynamic process that demands exquisite regulation. Follicles are restrained at the preantral stage until they are stimulated by the pituitary hormone follicle stimulating hormone (FSH). In response to FSH, GCs produce steroid and protein hormones and growth factors that regulate the hypothalamic/pituitary axis and uterine receptivity and promote oocyte maturation and development of the follicle to a preovulatory phenotype. We have shown that FSH promotes enhanced translation leading to an accumulation of the transcriptional factor hypoxia-inducible factor-1a (HIF1a) in GCs. HIF1a is normally absent under normoxia due to oxygen-dependent post-translational modifications that target it for proteosomal degradation, and accumulates only under hypoxic conditions. HIF1a, together with constitutively expressed HIF1b, form the heterodimeric transcriptional factor HIF1 that activates a hierarchy of target genes that permit cells to survive under reduced oxygen concentrations. The aims of this application test the overarching hypothesis that proper GC function and follicular maturation require a HIF1-responsive transcriptome optimally regulated by FSH and hypoxia. Support for these aims comes from our evidence that FSH-stimulated HIF1 activity appears to be necessary for induction of a diverse group of target genes that include vascular endothelial growth factor (Vegf), inhibin-a (officially Inha), luteinizing hormone/choriogonadotropin receptor (officially Lhcgr), and protein kinase A regulatory subunit RIIb (officially Prkar2b). With the exception of Vegf, these apparent HIF1 targets in GCs are distinct from other common targets of this ubiquitous transcriptional activator. Thus, elucidation of the functional role of HIF1 is important not only to GC maturation and fertility but may also contribute to our understanding of the misregulation of genes such as the Lhcgr seen in many different cancer cells that express high levels of HIF1a. Moreover, understanding how FSH signals to direct follicular maturation can translate into safer and more effective treatments of infertility and early pregnancy loss as well as new approaches for contraceptive drugs. PUBLIC HEALTH RELEVANCE: FSH signaling to mature follicles to the preovulatory phenotype is required for fertility. Understanding how FSH signals to direct follicular maturation can translate into safer and more effective treatments of infertility and early pregnancy loss as well as new approaches for contraceptive drugs.

描述（由申请人提供）：女性的生育能力需要卵母细胞、支持颗粒细胞（GC）和构成卵泡的卵泡膜细胞的受控成熟。支持卵泡成熟的营养物质和激素来自卵泡膜层的毛细血管；血管化终止于基底膜。因此，随着卵泡的生长，GC 暴露于越来越缺氧的环境中。卵泡生长是一个动态过程，需要精细的调节。卵泡在窦前阶段受到抑制，直到受到垂体激素卵泡刺激素（FSH）的刺激。为了响应 FSH，GC 产生类固醇和蛋白质激素以及生长因子，调节下丘脑/垂体轴和子宫容受性，并促进卵母细胞成熟和卵泡发育至排卵前表型。我们已经证明，FSH 促进翻译增强，导致转录因子缺氧诱导因子 1a (HIF1a) 在 GC 中积累。 HIF1a 在常氧条件下通常不存在，这是由于氧依赖性翻译后修饰靶向 HIF1a 进行蛋白体降解，并且仅在缺氧条件下积累。 HIF1a 与组成型表达的 HIF1b 一起形成异二聚体转录因子 HIF1，它激活一系列靶基因，使细胞能够在低氧浓度下生存。本申请的目的是测试一个总体假设，即适当的 GC 功能和卵泡成熟需要一个由 FSH 和缺氧最佳调节的 HIF1 响应转录组。对这些目标的支持来自于我们的证据，即 FSH 刺激的 HIF1 活性似乎对于诱导多种靶基因是必要的，这些靶基因包括血管内皮生长因子 (Vegf)、抑制素-a（官方称为 Inha）、黄体生成素/绒毛膜促性腺激素受体（正式名称为 Lhcgr）和蛋白激酶 A 调节亚基 RIIb（正式名称为 Prkar2b）。除了 Vegf 之外，GC 中这些明显的 HIF1 靶标与这种普遍存在的转录激活因子的其他常见靶标不同。因此，阐明 HIF1 的功能作用不仅对 GC 成熟和生育能力很重要，而且可能有助于我们理解基因的错误调节，例如在表达高水平 HIF1a 的许多不同癌细胞中观察到的 Lhcgr。此外，了解 FSH 如何发出引导卵泡成熟的信号可以转化为更安全、更有效的不孕不育和早期流产治疗方法以及避孕药物的新方法。公共健康相关性：FSH 信号将成熟卵泡转变成排卵前表型是生育能力所必需的。了解 FSH 信号如何指导卵泡成熟可以转化为更安全、更有效的不孕不育和早期流产治疗方法以及避孕药物的新方法。