Ad-vectors and Granulosa Cell Signaling

Ad 载体和颗粒细胞信号传导

• 批准号: 7099505
• 负责人: Anthony J Zeleznik
• 金额: $ 26.1万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7099505
• 关键词: Adenoviridae; RNase protection assay; activins; biological signal transduction; cell differentiation; cell proliferation; cyclic AMP; developmental genetics; drug interactions; enzyme activity; estrogens; follicle stimulating hormone; gene expression; genetic regulation; granulosa cell; hormone regulation /control mechanism; laboratory rat; luteinizing hormone; messenger RNA; nuclear receptors; phosphatidylinositol 3 kinase; progesterone; radioimmunoassay; serine threonine protein kinase; steroid hormone biosynthesis; tissue /cell culture; transfection /expression vector

DESCRIPTION (provided by applicant): Although progress has been made in understanding the individual roles of follicle stimulating hormone (FSH) an luteinizing hormone (LH) in governing the process of preovulatory folliculogenesis, a major question which remains unanswered is the elucidation of the intracellular signaling pathways utilized by FSH and LH in their regulation of the complex pattern of gene expression that occurs during follicular development. The research proposed herein will make use of replication-defective adenovirus vectors to explore the signaling pathways involved in granulosa cell proliferation and differentiation. Because these vectors infect granulosa cells and direct the expression of proteins with high efficiency it is possible to express constitutively activated as well as dominant-negative signaling proteins and directly assess their influences on granulosa cell steroidogenesis and cAMP production as well as the expression of mRNAs that encode for differentiation and proliferation-associated proteins. Aim 1will elucidate the mechanism by which FSH activates the PI3- kinase/PKB intracellular signaling pathway in granulosa cells. Aim 2 will identify downstream effectors of PI3-kinase/ PKB signaling system in granulosa cells that govern granulosa cell differentiation. Aim 3 will determine if LRH-1 and/or SF-1 are necessary and sufficient for cAMP-induced expression of mRNAs for enzymes involved in progesterone, but not estrogen, production during granulosa cell differentiation and to determine if SF-1 and LRH-1 are equally effective in this regard and Aim 4 will identify the intracellular signaling pathways responsible for the synergistic role of activin in FSH-stimulated granulosa cell proliferation. Understanding the intracellular signaling pathways utilized by the FSH receptor and the LH receptor will be instrumental for the development of the next generation of contraceptives as well as the next generation of "fertility drugs" in which FSH and LH signaling pathways are targeted directly. In addition, results may provide new information regarding signaling pathways that may be affected in PCOS.

描述(申请人提供)：虽然在了解卵泡刺激素(FSH)和黄体生成素(LH)在控制排卵前卵泡发生过程中的个体作用方面已经取得了进展，但一个尚未回答的主要问题是FSH和LH在调节卵泡发育过程中复杂的基因表达模式时所使用的细胞内信号通路的阐明。本研究将利用复制缺陷型腺病毒载体探索颗粒细胞增殖和分化的信号转导途径。由于这些载体可以高效地感染颗粒细胞并指导蛋白质的表达，因此可以表达结构性激活的和显性负相关的信号蛋白，并直接评估它们对颗粒细胞类固醇合成和cAMP产生的影响，以及编码分化和增殖相关蛋白的mRNAs的表达。目的1阐明卵泡刺激素(FSH)激活颗粒细胞内PI3K/PKB信号通路的机制。目的2将确定颗粒细胞中控制颗粒细胞分化的PI3-激酶/PKB信号系统的下游效应因子。目的3将确定LRH-1和/或SF-1是否是cAMP诱导颗粒细胞分化过程中孕酮(而不是雌激素)相关酶表达的必要条件和充分条件，并确定SF-1和LRH-1在这方面是否同样有效，目的4将确定激活素在FSH刺激的颗粒细胞增殖中协同作用的细胞内信号通路。了解卵泡刺激素受体和促黄体生成素受体所利用的细胞内信号通路将有助于开发下一代避孕药以及直接针对卵泡刺激素和促黄体生成素信号通路的下一代生育药物。此外，结果可能提供有关多囊卵巢综合征可能受影响的信号通路的新信息。