AGE-REALTED CHANGES IN GONADOTROPIN GLYCOSYLATION AND FUNCTION

促性腺激素糖基化和功能的年龄相关变化

• 批准号: 7651594
• 负责人: GEORGE R BOUSFIELD
• 金额: $ 27.21万
• 依托单位: WICHITA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-15 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7651594
• 关键词: Affinity; Age; Aging; Attenuated; Binding; Biochemical Markers; Biological; Biological Assay; Data; Early Diagnosis; Estrogen Replacement Therapy; Estrogens; Exhibits; Face; Feedback; Fertility; Follicle Stimulating Hormone Receptor; Future; Genitourinary system; Goals; Gonadotropins; Human; Human Follicle Stimulating Hormone; Hybrids; Impairment; Individual; Infertility; Lead; Mass Spectrum Analysis; Measurement; Measures; Menopause; Methods; Minor; Morbidity - disease rate; Outcome; Ovarian; Ovary; Patients; Pattern; Perimenopause; Pharmaceutical Preparations; Physiological; Pituitary Gland; Polysaccharides; Population; Postmenopause; Preparation; Primates; Principal Investigator; Procedures; Protein Isoforms; Rattus; Receptor Activation; Recombinants; Regulation; Relative (related person); Reporting; Research; Role; Sampling; Sialic Acids; Site; Steroid biosynthesis; Structure; Symptoms; Techniques; Testing; Time; Urine; Uterine hemorrhage; Variant; Vasomotor; Western Blotting; Woman; Work; age related; assisted reproduction; base; bone mass; experience; glycosylation; granulosa cell; improved; innovation; method development; mullerian-inhibiting hormone; novel; older women; pituitary gonadal axis; programs; protein protein interaction; psychologic; receptor; receptor binding; reproductive; urinary

Our long-term goal is to determine how gonadotropins function. Gonadotropins require N-glycans to fully activate their cognate receptors even though high affinity binding involves only protein-protein interactions. Gonadotropins are used to treat infertility, but become progressively less effective as patients age. Urinary and recombinant hFSH preparations lack a novel, highly active glycoform, di-glycosylated hFSH, that possesses only a subunit N-glycans and which predominates in pituitaries of young women. The relative abundance of di-glycosylated and tetra-glycosylated hFSH appear to be physiologically regulated. This proposal will define these changes and study possible mechanisms to explain differences in activity via 2 specific aims. 1. Characterize cyclic- and age-related changes in hFSH glycoform abundance. First, we will measure urinary FSH glycoform abundance in young healthy cycling women. Our working hypothesis is that ovarian feedback will alter FSH glycoform abundance. Second, we will evaluate glycoform abundance during the perimenopausal period, as the timing of the shift in glycoform abundance is unknown. One hypothesis is that FSH glycoform abundance changes gradually during the perimenopausal period, thereby contributing to declining fertility. An alternative hypothesis is that glycoform abundance changes abruptly at the menopausal transition. Finally, we will examine glycoform abundance in post-menopausal women and compare individuals receiving estrogen replacement therapy with those receiving none. Our working hypothesis is estrogen selectively inhibits hFSHfi glycosylation. 2. Identify a and /? subunit glycosylation patterns modulating FSH receptor-binding and FSH-stimulated steroidogenesis. We will compare receptor-binding and steroidogenic activities of di- and tetra-glycosylated hFSH. Our hypothesis is that di-glycosylated hFSH on rate is faster than tetra-glycosylated hFSH. The relationship of FSHR binding and steroidogenic activity will be compared. Our working hypothesis is that di-glycosylated hFSH will exhibit greater biological activity than predicted by its significatly higher receptor-binding activity. This project is centered around glycoforms readily identified by western blotting and lays the ground for future studies by contributing to the development of methods to rapidly characterize FSH glycosylation. A perimenopausal marker and more effective preparations for treating infertility are potential practical outcomes of this project.

我们的长期目标是确定促性腺激素如何发挥作用。促性腺激素需要 N-聚糖 即使高亲和力结合仅涉及蛋白质-蛋白质，也能完全激活其同源受体 互动。促性腺激素用于治疗不孕症，但随着患者的治疗效果逐渐减弱 年龄。尿和重组 hFSH 制剂缺乏一种新型的、高活性的糖型，即二糖基化 hFSH， 它只含有一个 N-聚糖亚基，在年轻女性的垂体中占主导地位。这 二糖基化和四糖基化 hFSH 的相对丰度似乎受到生理调节。 该提案将定义这些变化并研究可能的机制来解释活动的差异 2具体目标。 1. 表征 hFSH 糖型丰度的周期性和年龄相关变化。 首先，我们将测量年轻健康骑行女性的尿 FSH 糖型丰度。我们的工作 假设卵巢反馈会改变 FSH 糖型丰度。其次，我们会评估 围绝经期期间的糖型丰度，因为糖型丰度转变的时间是 未知。一种假设是 FSH 糖型丰度在 围绝经期，从而导致生育能力下降。另一种假设是 糖型丰度在绝经过渡期突然变化。最后，我们将检查糖型 绝经后妇女的丰度并比较接受雌激素替代治疗的个体 与那些没有收到的人。我们的工作假设是雌激素选择性抑制 hFSHfi 糖基化。 2. 识别一个和/？亚基糖基化模式调节 FSH 受体结合和 FSH 刺激类固醇生成。我们将比较二元的受体结合和类固醇生成活性 和四糖基化 hFSH。我们的假设是二糖基化 hFSH 的作用速度比 四糖基化 hFSH。将比较 FSHR 结合和类固醇生成活性的关系。我们的 工作假设是二糖基化 hFSH 将表现出比其预测更高的生物活性 显着更高的受体结合活性。 该项目以通过蛋白质印迹法轻松识别的糖型为中心，并奠定了基础 通过开发快速表征 FSH 糖基化的方法，为未来的研究做出贡献。一个 围绝经期标志物和更有效的治疗不孕症的制剂具有潜在的实用性 该项目的成果。