Ad-vectors and Granulosa Cell Signaling

Ad 载体和颗粒细胞信号传导

基本信息

批准号：
7271162
负责人：
Anthony J Zeleznik
金额：
$ 25.34万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-01 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7271162
关键词：
1-Phosphatidylinositol 3-Kinase Activins Address Adopted Affect Aromatase Cell Differentiation process Cell Proliferation Cell physiology Complex Contraceptive Agents Cyclic AMP Cyclic AMP-Dependent Protein Kinases Development Differentiation and Growth Dominant-Negative Mutation Enzymes Estradiol Estrogens Female Fertility Agents Follicle Stimulating Hormone Follicle Stimulating Hormone Receptor Funding Gene Expression Homologous Gene Hormone Receptor Individual LH Receptors Liver Luteinizing Hormone Mediating Nuclear Orphan Receptor Ovarian Ovarian Follicle PCNA gene Pathway interactions Pattern Phosphoinositide-3-Kinase, Catalytic, Gamma Polypeptide Play Process Production Progesterone Proteins Proto-Oncogene Proteins c-akt Regulation Reproduction Research Role Signal Pathway Signal Transduction Signaling Protein Specificity Steroid biosynthesis System cell type cyclin D2 defective adenoviral vector folliculogenesis granulosa cell in vivo inhibitor/antagonist next generation novel protein expression receptor vector

项目摘要

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）：虽然在了解促卵泡激素（FSH）和促黄体生成激素（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，但仍未得到解答的一个主要问题是阐明细胞内促黄体生成素（LH）的作用。 FSH 和 LH 在调节卵泡发育过程中发生的复杂基因表达模式时利用的信号通路。本文提出的研究将利用复制缺陷型腺病毒载体来探索颗粒细胞增殖和分化所涉及的信号通路。由于这些载体感染颗粒细胞并高效指导蛋白质的表达，因此可以表达组成型激活的信号蛋白以及显性失活信号蛋白，并直接评估它们对颗粒细胞类固醇生成和 cAMP 产生以及 mRNA 表达的影响。编码分化和增殖相关蛋白。目标 1 将阐明 FSH 激活颗粒细胞中 PI3-激酶/PKB 细胞内信号通路的机制。目标 2 将鉴定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。目标 3 将确定 LRH-1 和/或 SF-1 对于 cAMP 诱导的 mRNA 表达是否是必要且充分的，这些酶涉及颗粒细胞分化过程中黄体酮（而非雌激素）的产生，并确定 SF-1 和 LRH-1 是否是必要且充分的。 1 在这方面同样有效，目标 4 将确定负责激活素在 FSH 刺激的颗粒细胞增殖中协同作用的细胞内信号传导途径。了解 FSH 受体和 LH 受体利用的细胞内信号传导途径将有助于开发下一代避孕药以及直接针对 FSH 和 LH 信号传导途径的下一代“生育药物”。此外，结果可能提供有关 PCOS 中可能受影响的信号通路的新信息。