Molecular Basis of BMP-15 Function

BMP-15 功能的分子基础

• 批准号: 8097123
• 负责人: SHUNICHI SHIMASAKI
• 金额: $ 8.06万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 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 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8097123
• 关键词: Antral; Applications Grants; Binding; Biochemical; Biological; Biological Process; Biology; Cell Proliferation; Cell physiology; Cells; Characteristics; Data; Defect; Development; Exhibits; Female; Fertility; Genes; Glean; Goals; Golgi Apparatus; Grant; Growth Differentiation Factor 9; Growth Factor; Growth and Development function; Human; Incidence; Infertility; Knock-out; Knowledge; Laboratories; Ligands; Mass Spectrum Analysis; Mitosis; Molecular; Mus; Mutation; Oocytes; Ovarian; Ovarian Follicle; Ovary; Ovulation; Patients; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Play; Positioning Attribute; Post-Translational Protein Processing; Premature Ovarian Failure; Process; Progress Reports; Property; Proteins; Proteomics; Publishing; Recombinants; Regulation; Rodent; Role; Signal Transduction Pathway; Staging; Structure; Techniques; Testing; Time; Translating; Treatment Protocols; Woman; Work; base; bone morphogenetic protein 15; bone morphogenetic protein receptor type II; casein kinase; folliculogenesis; granulosa cell; member; mutant; novel; overexpression; prevent; public health relevance; receptor; research study; treatment strategy

DESCRIPTION (provided by applicant): The goal of this renewal grant is to translate our pioneering studies on the biology of bone morphogenetic protein-15 (BMP-15) in rodents to the human. The rationale underpinning the grant is that mutations in the bmp15 gene cause infertility in women due to defects in folliculogenesis. BMP-15 has a critical physiological role in regulating granulosa cell proliferation and differentiation in women. At present, almost nothing is known about how BMP-15 acts to control human granulosa cell function. We have recently identified posttranslational modifications in the primary structure of recombinant human BMP-15 (rhBMP-15) by proteomics analysis, most notably a phosphorylation in the mature region. Importantly, phosphorylation is essential for the bioactivity of rhBMP-15. Interestingly, de-phosphorylated rhBMP-15 can negate the bioactivity of phosphorylated rhBMP-15. Moreover, de-phosphorylated rhBMP-15 also inhibits the bioactivity of recombinant human growth and differentiation factor-9 (rhGDF-9) and rhBMP-7 that share a common BMP type II receptor with rhBMP-15. However, de-phosphorylated rhBMP-15 does not impair the bioactivity of rhActivin A, which shares neither type I and type II receptors with rhBMP-15. The hypothesis that emerges from these novel findings is that protein phosphorylation of BMP-15 may represent a primary physiological determinant for ovarian function and fertility in women, and that the non-phosphorylated BMP-15 may be a functional antagonist for not only BMP-15 but also for other selective members of the TGF-? superfamily. Four Specific Aims are proposed to test these hypotheses. Aim 1: Determine the biological function of rhBMP-15 in human granulosa cells at defined stages of follicle development. Aim 2: Establish the role of phosphorylation in the bioactivity of rhBMP-15 in human granulosa cells. Aim 3: Investigate the molecular and cellular mechanisms by which non-phosphorylated rhBMP-15 antagonizes broadly the activity of selective BMP/GDF ligands. Aim 4: Determine the biochemical characteristics and the biological functions of the BMP-15 mutants identified in premature ovarian failure (POF) patients and explore to what extent the mutations are functionally associated with the phenotype of POF patients. The proposed studies should provide significant advances in understanding the regulatory mechanisms of follicle growth and development by the oocyte-specific factor, BMP-15, and could hold promise for new strategies for the treatment of infertility and regulation of fertility in women. PUBLIC HEALTH RELEVANCE: Bone Morphogenetic Protein-15 (BMP-15) is a critical growth factor that controls ovarian follicle growth and development, leading to the normal ovulation, thus, female fertility. In this grant application, we propose to uncover the biological functions of BMP-15 and the underlying molecular mechanism in the human ovary. The proposed studies should provide significant advances in understanding the regulatory mechanisms of follicle growth and development by BMP-15, and ultimately develop new regimens for the treatment of infertility and regulation of fertility in women.

描述（由申请人提供）：本续期资助的目标是将我们在啮齿动物骨形态发生蛋白-15 （BMP-15）生物学方面的开创性研究转化为人类。这项资助的基本原理是，bmp15基因的突变会导致女性因卵泡发育缺陷而不孕。BMP-15在调节女性颗粒细胞增殖和分化中具有重要的生理作用。目前，关于BMP-15如何控制人颗粒细胞功能几乎一无所知。我们最近通过蛋白质组学分析确定了重组人BMP-15 （rhBMP-15）初级结构的翻译后修饰，最明显的是成熟区磷酸化。重要的是，磷酸化对rhBMP-15的生物活性至关重要。有趣的是，去磷酸化的rhBMP-15可以否定磷酸化的rhBMP-15的生物活性。此外，去磷酸化的rhBMP-15还抑制了重组人生长与分化因子-9 （rhGDF-9）和rhBMP-7的生物活性，这两种细胞与rhBMP-15具有共同的BMP II型受体。然而，去磷酸化的rhBMP-15不会损害rhActivin A的生物活性，rhBMP-15与rhBMP-15既不共享I型受体，也不共享II型受体。从这些新发现中产生的假设是，BMP-15的蛋白磷酸化可能代表了女性卵巢功能和生育能力的主要生理决定因素，而非磷酸化的BMP-15可能不仅是BMP-15的功能性拮抗剂，也可能是TGF-？总科。提出了四个具体目标来检验这些假设。目的1：确定rhBMP-15在卵泡发育特定阶段的人颗粒细胞中的生物学功能。目的2：建立磷酸化在人颗粒细胞中rhBMP-15生物活性中的作用。目的3：研究非磷酸化的rhBMP-15广泛拮抗选择性BMP/GDF配体活性的分子和细胞机制。目的4：确定卵巢早衰（POF）患者中发现的BMP-15突变体的生化特征和生物学功能，并探讨这些突变在多大程度上与POF患者的表型存在功能关联。这些研究将为了解卵泡生长和发育的卵泡特异性因子BMP-15的调节机制提供重大进展，并有望为治疗不孕症和调节女性生育能力提供新的策略。公共卫生相关性：骨形态发生蛋白-15 （BMP-15）是一种关键的生长因子，控制卵巢卵泡生长和发育，导致正常排卵，从而提高女性生育能力。在本次拨款申请中，我们建议揭示BMP-15的生物学功能及其在人类卵巢中的潜在分子机制。建议的研究将为了解BMP-15对卵泡生长发育的调节机制提供重大进展，并最终为治疗不孕症和调节女性生育能力提供新的方案。