Function of RUNX transcription factors in COCs

RUNX转录因子在COC中的功能

• 批准号: 8147698
• 负责人: MISUNG JO
• 金额: $ 24.24万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-24 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8147698
• 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

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. PUBLIC HEALTH RELEVANCE: Proper maturation of COCs is requisite for successful ovulation and fertilization. Appropriate COC expansion induced by the preovulatory gonadotropin surge is directly linked to oocyte quality and developmental competence. There are still huge gaps in our understanding of how the preovulatory gonadotropin surge induces the maturation process in COCs. Previous studies and our preliminary data suggest that RUNX transcription factor(s) plays a critical role in cumulus expansion by regulating the transcription of specific periovulatory cumulus genes. Building upon these findings, the present proposal will determine the molecular/cellular mechanism(s) involved in COC expansion by elucidating the functional role(s) of RUNX transcription factors during maturation of COCs. Information derived from the present proposal will provide new insight into the mechanisms involved in COC maturation. Such knowledge can be applied for promoting and inhibiting this critical facet of ovarian physiology, thereby allowing us to better manage 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生殖成功至关重要。我们对这一成熟过程缺乏完整的了解，限制了我们管理体内生育能力的能力，并阻碍了体外受精和体外成熟等辅助生殖技术的实施。我们实验室和其他人最近的研究已经开始揭示核转录因子 RUNX1 和 RUNX2 的一个小家族，它们是参与 COC 扩增的关键转录调节因子。我们的初步数据表明，Runx1 和 Runx2 的表达在排卵周围卵泡的卵丘细胞中高度诱导。我们还发现RUNX1和RUNX2调节壁颗粒细胞和/或卵丘细胞中Ptgs2（前列腺素生物合成中的限速酶）和Hapln1（卵丘基质的稳定剂）的表达。已知这两种基因产物在 COC 扩增中发挥着至关重要的作用。最重要的是，我们的初步研究表明，卵丘细胞中显性失活 RUNX 的过度表达可阻止 COC 扩增。基于这些新发现，我们假设 RUNX1/2 是促性腺激素激增诱导 COC 扩增所必需的重要转录调节因子。该假设将通过首先证明 Runx1/2 表达在积云细胞中的功能意义来检验（目标#1）。其次，我们将确定 RUNX1 和 RUNX2 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 Runx1 和 Runx2 表达的调节机制（目标#3）。这 3 个目标将使用大鼠 COC 进行研究。为了确保从大鼠研究中获得的数据可以转化为人类，RUNX1/2 及其靶基因表达谱也将使用整个排卵期收集的人类卵丘细胞进行验证。我们还将使用从接受 IVF 的女性获得的 COC，检查 RUNX1/2 的卵丘表达及其靶基因与卵母细胞质量、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。从拟议研究中获得的信息不仅将增进我们对 COC 成熟机制的理解，而且有助于未来的转化/临床应用，从而改善体内和体外生育管理。 公众健康相关性：COC 的适当成熟是成功排卵和受精的必要条件。排卵前促性腺激素激增诱导的适当 COC 扩张与卵母细胞质量和发育能力直接相关。我们对排卵前促性腺激素激增如何诱导 COC 成熟过程的理解仍然存在巨大差距。之前的研究和我们的初步数据表明，RUNX 转录因子通过调节特定排卵期卵丘基因的转录，在卵丘扩张中发挥着关键作用。基于这些发现，本提案将通过阐明 RUNX 转录因子在 COC 成熟过程中的功能作用来确定参与 COC 扩增的分子/细胞机制。从本提案中获得的信息将为 COC 成熟所涉及的机制提供新的见解。这些知识可用于促进和抑制卵巢生理学的这一关键方面，从而使我们能够更好地管理体内和体外的生育能力。