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Function of RUNX transcription factors in COCs

RUNX转录因子在COC中的功能

基本信息

批准号：
8484756
负责人：
MISUNG JO
金额：
$ 23万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-24 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8484756
关键词：
Adenoviruses Anabolism Animal Model Cells Collecting Cell Competence Complex Data Development Dominant-Negative Mutation Embryonic Development Enzymes Female Fertility Fertilization Fertilization in Vitro Foundations Future GDF9 gene Gene Expression Gene Expression Profile Gene Targeting Genes Genetic Transcription Goals Gonadotropins Hormonal Human In Vitro Knowledge Laboratories Light Link Mediating Mediator of activation protein Molecular Molecular Profiling Mus Nuclear Family Oocytes Ovarian Ovulation Peptides Physiology Pilot Projects Play Process Prostaglandins Proteins RUNX1 gene Rattus Regulation Role Signal Transduction Small Interfering RNA Testing Woman base clinical application combinatorial cytokine granulosa cell implantation improved in vivo insight novel promoter public health relevance reproductive success success time interval transcription factor

项目摘要

DESCRIPTION (provided by applicant): The overall goal of the present proposal is to determine the molecular/cellular mechanism(s) involved in the maturation process of cumulus oocyte complexes (COCs). Proper maturation of the COC is critical to reproductive success in vivo and in vitro. Our lack of a complete understanding of this maturation process restricts our ability to manage fertility in vivo and hinders implementation of ART such as in vitro fertilization and in vitro maturation. Recent studies by our laboratory and others have begun to shed light on a small family of nuclear transcription factors, RUNX1 and RUNX2, as key transcriptional regulators involved in COC expansion. Our preliminary data demonstrated that Runx1 and Runx2 expression is highly induced in cumulus cells of periovulatory follicles. We also found that RUNX1 and RUNX2 regulate the expression of Ptgs2 (a rate-limiting enzyme in the biosynthesis of prostaglandins) and Hapln1 (a stabilizer of cumulus matrix) in mural granulosa cells and/or cumulus cells. Both gene products are known to play crucial roles in COC expansion. Most importantly, our pilot study demonstrated that over-expression of dominant negative RUNX in cumulus cells blocked COC expansion. Based on these novel findings, we hypothesized that RUNX1/2 are essential transcriptional regulators necessary for the gonadotropin surge-induced COC expansion. This hypothesis will be tested by first demonstrating the functional significance of Runx1/2 expression in cumulus cells (Aim #1). Secondly, we will determine the regulation action(s) of RUNX1 and RUNX2 on their target genes (Aim #2). Next, we will determine the regulatory mechanism(s) of Runx1 and Runx2 expression in cumulus cells (Aim #3). These 3 Aims will be studied using rat COCs. To insure that the data obtained from rat studies is translatable to humans, RUNX1/2 and their target gene expression profile will be also verified using human cumulus cells collected throughout the periovulatory period. We will also examine the correlation of the cumulus expression of RUNX1/2 and their target genes to oocyte quality, fertilization, and embryo development using COCs obtained from women undergoing IVF (Aim #4). These human studies will serve as a foundation for future translational/clinical application. The information obtained from the proposed studies will not only advance our understanding of the mechanism involved in COC maturation, but also be instrumental for future translational/clinical application(s), thus leading to improved management of fertility in vivo and in vitro.

描述（由申请人提供）：本提案的总体目标是确定卵丘卵母细胞复合体（COC）成熟过程中涉及的分子/细胞机制。COC的适当成熟对于体内和体外繁殖成功至关重要。我们对这种成熟过程缺乏完整的了解，限制了我们在体内管理生育能力的能力，并阻碍了ART的实施，如体外受精和体外成熟。我们实验室和其他人最近的研究已经开始揭示一个小家族的核转录因子，RUNX 1和RUNX 2，作为关键的转录调控参与COC扩增。我们的初步数据表明，Runx 1和Runx 2的表达是高度诱导的卵丘细胞的排卵期卵泡。我们还发现，RUNX 1和RUNX 2调节Ptgs 2的表达（一种限速酶的生物合成的三尖杉酯碱）和Hapln 1（稳定剂的卵丘基质）在壁颗粒细胞和/或卵丘细胞。已知这两种基因产物在COC扩增中起关键作用。最重要的是，我们的初步研究表明，卵丘细胞中显性负RUNX的过度表达阻断了COC扩增。基于这些新的发现，我们假设RUNX 1/2是促性腺激素激增诱导的COC扩增所必需的重要转录调节因子。将通过首先证明卵丘细胞中Runx 1/2表达的功能意义来检验该假设（目的#1）。其次，我们将确定RUNX 1和RUNX 2对其靶基因的调节作用（目标#2）。接下来，我们将确定卵丘细胞中Runx 1和Runx 2表达的调控机制（目标#3）。将使用大鼠COC研究这3个目的。为了确保从大鼠研究中获得的数据可转化为人类，还将使用整个排卵期收集的人卵丘细胞验证RUNX 1/2及其靶基因表达谱。我们还将使用从接受IVF的妇女中获得的COC来检查RUNX 1/2及其靶基因的卵丘表达与卵母细胞质量、受精和胚胎发育的相关性（目标#4）。这些人体研究将作为未来转化/临床应用的基础。从拟议的研究中获得的信息不仅将促进我们对COC成熟机制的理解，而且还有助于未来的翻译/临床应用，从而改善体内和体外生育力的管理。