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（由申请人提供）：尽管在理解卵泡刺激激素（FSH）的个体作用方面取得了进展 发展。本文提出的研究将利用复制缺陷的腺病毒载体探索与颗粒细胞增殖和分化有关的信号传导途径。由于这些载体感染了颗粒细胞并以高效率指导蛋白质的表达，因此有可能表达组成型激活以及显性阴性信号蛋白，并直接评估它们对颗粒类细胞类固醇生成和cAMP产生的影响，以及编码分化和扩增蛋白质的MRNA的表达。 AIM 1WILL阐明了FSH激活颗粒细胞中PI3-激酶/PKB细胞内信号通路的机制。 AIM 2将识别控制颗粒细胞分化的颗粒细胞中PI3-激酶/ PKB信号系统的下游效应子。 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.了解FSH受体和LH受体使用的细胞内信号通路将有助于下一代避孕药的发展以及下一代的“生育药物”，其中FSH和LH信号传导途径是直接针对的。此外，结果可能会提供有关PCOS可能影响的信号通路的新信息。