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）：本次更新资助的目标是将我们对啮齿动物骨形态发生蛋白-15（BMP-15）生物学的开创性研究转化为人类。支持这项资助的基本原理是，bmp 15基因突变会导致女性不孕，这是由于卵泡发育缺陷。BMP-15在调节女性颗粒细胞增殖和分化中具有关键的生理作用。目前，关于BMP-15如何控制人类颗粒细胞功能几乎一无所知。我们最近已经确定了翻译后修饰的重组人BMP-15（rhBMP-15）的一级结构的蛋白质组学分析，最值得注意的是在成熟区域的磷酸化。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rhBMP-15还抑制重组人生长和分化因子-9（rhGDF-9）和rhBMP-7的生物活性，rhGDF-9和rhBMP-7与rhBMP-15共享共同的BMP II型受体。然而，去磷酸化rhBMP-15不会损害rhActivin A的生物活性，rhActivin A与rhBMP-15不共享I型和II型受体。从这些新的发现出现的假设是，BMP-15的蛋白磷酸化可能是一个主要的生理决定因素，卵巢功能和生育能力的妇女，和非磷酸化的BMP-15可能是一个功能性拮抗剂，不仅BMP-15，但也为其他选择性成员的TGF-？超家族提出了四个具体目标来检验这些假设。目的1：确定rhBMP-15在人卵泡发育的特定阶段的颗粒细胞中的生物学功能。目的2：研究磷酸化对rhBMP-15在人卵巢颗粒细胞中生物活性的影响。目标3：研究非磷酸化rhBMP-15广泛拮抗选择性BMP/GDF配体活性的分子和细胞机制。目标4：确定在卵巢早衰（POF）患者中鉴定的BMP-15突变体的生化特征和生物学功能，并探索突变在多大程度上与POF患者的表型功能相关。拟议的研究应提供重大进展，了解卵泡生长和发育的调控机制的卵母细胞特异性因子，BMP-15，并可能持有承诺的新策略，为治疗不孕症和调节妇女的生育能力。公共卫生关系：骨形态发生蛋白-15（BMP-15）是一种重要的生长因子，控制卵泡的生长和发育，导致正常排卵，从而提高女性生育力。在这项资助申请中，我们计划揭示BMP-15在人类卵巢中的生物学功能和潜在的分子机制。拟议的研究将在理解BMP-15对卵泡生长和发育的调节机制方面取得重大进展，并最终开发出治疗不孕症和调节女性生育力的新方案。