Functional Significance of the HIF1 Transcriptome in Granulosa Cells

颗粒细胞中 HIF1 转录组的功能意义

• 批准号: 7942600
• 负责人: Mary E Hunzicker-Dunn
• 金额: $ 30.51万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 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/7942600
• 关键词: 1-Phosphatidylinositol 3-Kinase; Basement membrane; Biological Assay; Blood capillaries; Cell Culture Techniques; Cell Extracts; 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

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控制卵母细胞的成熟，支撑颗粒细胞（GCS）和构成卵巢卵泡的theca细胞。支持卵泡成熟的营养和激素来自毛细血管的毛细血管；血管化终止于基底膜。因此，随着卵泡的生长，GC暴露于越来越多的低氧条件下。卵泡生长是一个动态过程，需要精致的调节。卵泡在尿下阶段受到约束，直到被垂体激素卵泡刺激激素（FSH）刺激。为了响应FSH，GC会产生类固醇和蛋白质激素以及生长因子，这些激素和生长因子调节下丘脑/垂体轴和子宫接受能力，并促进卵母细胞的成熟以及卵泡发展为卵巢型表型。我们已经表明，FSH促进了增强的翻译，从而导致转录因子缺氧诱导因子1a（HIF1A）在GC中的积累。由于氧气依赖性的翻译后修饰，将其用于蛋白质体降解，并且仅在低氧条件下积累，因此HIF1A通常不存在于常氧中。 HIF1A与组成型表达的HIF1B一起形成了异二聚体转录因子HIF1，该因子激活了靶基因的层次结构，该靶基因允许细胞在降低的氧气浓度下生存。该应用程序的目的检验了总体假设，即适当的GC功能和卵泡成熟需要由FSH和缺氧最佳调节的HIF1响应转录组。 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）。除VEGF外，GC中的这些明显的HIF1靶标不同于此无处不在的转录激活剂的其他常见靶标。因此，阐明HIF1的功能作用不仅对GC的成熟和生育重要性很重要，而且还可能有助于我们理解基因的不正直，例如在许多表达高水平HIF1A的癌细胞中看到的LHCGR。此外，了解FSH信号如何指导卵泡成熟度如何转化为对不育和早期妊娠丧失的更安全，更有效的治疗方法，以及避孕药的新方法。 公共卫生相关性：对成熟卵泡的FSH信号传导至排卵前表型才能生育。 了解指导卵泡成熟的信号如何转化为不育和早期妊娠丧失的更安全，更有效的治疗方法，以及避孕药的新方法。