Elucidating the Role of YAP and TAZ in the Aging Human Ovary

阐明 YAP 和 TAZ 在人类卵巢衰老中的作用

• 批准号: 10722368
• 负责人: JOHN S DAVIS
• 金额: $ 42.21万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-07 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10722368
• 关键词: ATAC-seq; Address; Affect; Age; Aging; Architecture; Automobile Driving; Binding; Biochemical; Bioinformatics; Candidate Disease Gene; Categories; Cattle; Cell Cycle; Cell Fate Control; Cell Line; Cell Proliferation; Cell physiology; Childbirth; Chromatin; Clinical; Communication; Complex; Contraceptive methods; DNA; Data; Development; Diagnosis; Down-Regulation; Education; Elderly; Endocrine; Enzymes; Estrogens; Event; Female infertility; Fertility; Fertilization in Vitro; Follicle Stimulating Hormone; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Transcription; Genomics; Goals; Gonadotropins; Growth; Homeostasis; Human; Infertility; Irregular Menstruation; Knowledge; Mediating; Medical; Menopause; Molecular; Mothers; Mus; Natural Fertility; Nuclear; Occupations; Oocytes; Organ Size; Ovarian; Ovarian Follicle; Ovarian aging; Ovary; Pathway interactions; Patients; Pattern; Perimenopause; Play; Polycystic Ovary Syndrome; Procedures; Process; Proliferating; Proteins; Protocols documentation; Publishing; Reporting; Research; Role; Sampling; Serum; Signal Pathway; Signal Transduction; Small Interfering RNA; Social Change; Steroid biosynthesis; Techniques; Time; Transcriptional Regulation; Validation; Withdrawal; Woman; advanced maternal age; age related; connective tissue growth factor; diminished ovarian reserve; experience; fertility preservation; gain of function; genetic signature; granulosa cell; health disparity; improved; infertility treatment; innovation; insight; mechanical signal; mullerian-inhibiting hormone; multiple omics; novel; nuclease; older women; oocyte maturation; paracrine; patient response; programs; reproductive; response; senescence; social structure; success; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; trend; tumorigenesis; young woman

PROJECT SUMMARY Ovarian senescence begins about age 30 and over the next 10 to 15 years is manifested by infertility. An expanded knowledge of the underlying molecular mechanisms that dictate granulosa cell function will facilitate our understanding of the health disparities of age and infertility. Over the past half century, the number of first- time mothers of advanced maternal aged women 35 years and older has increased more than five-fold. With more women entering the workforce, changes in social structure, advanced education and increased use of contraception, this trend will likely continue to increase. There is a constant bi-directional communication between the oocytes and granulosa cells of the follicle, which is required for development and maturation of high-quality follicles. The Hippo signaling is known to regulate organ size. Our published results established the role of Hippo signaling, and its downstream effector molecules, YAP and TAZ, as critical regulators of granulosa cell function and fertility. Notably, our preliminary analysis identified YAP/TAZ activity to be downregulated in advanced aged women (≥ 40 years) undergoing IVF and having low oocyte yields. We hypothesize that YAP and TAZ orchestrate crucial transcriptional events that regulate development of high-quality follicles. However, there is a gap in knowledge in understanding the actual mechanism of how these transcription factors regulate granulosa cell function, especially in aging women undergoing IVF. The overarching goal of this proposal is to understand YAP- and TAZ-mediated transcription and chromatin architecture in granulosa cells of women with advanced age. A drawback of using primary patient granulosa samples are low sample numbers which is a major roadblock to progress. The major innovation of this proposal is the ability to simultaneous profile chromatin and transcription regulation in the same patient sample thus providing greater in-depth understanding of the role of YAP and TAZ transcription factors in granulosa cell function. Our research strategy includes two specific aims. In Specific Aim 1. We will define the transcriptional targets of YAP and TAZ in granulosa cells by using CUT&RUN, a micrococal nuclease technique which can isolate specific protein-DNA complexes and distinguish direct TF binding from low numbers of granulosa cells. Our analysis includes granulosa cells obtained from IVF patients with poor response (< 6 oocytes retrieved) and good response (> 17 oocytes retrieved) in women of advanced maternal age (≥ 40 yrs old). We will also employ ATAC-seq to determine chromatin accessibility in these patient samples. In Aim 2, we will modulate YAP and TAZ in human granulosa cells and define their resultant transcriptomic profile. Genes identified in this study will be functionally validated by analysis of granulosa cell proliferation, differentiation, and senescence. These novel insights will help to better understand age-related decline in fertility and the role of the Hippo signaling pathway effectors YAP and TAZ in the ovary.

项目摘要 卵巢敏感始于大约30岁，在接下来的10至15年中，不孕症表现出来。一个 扩展了对决定颗粒细胞功能的基本分子机制的知识，将有助于 我们对年龄和不育的健康分布的理解。在过去的半个世纪中，首先 35岁及以上的高级母性女性的时代母亲增加了五倍以上。和 越来越多的妇女进入劳动力，社会结构的变化，高级教育和增加的使用 避孕，这种趋势可能会继续增加。有持续的双向通信 在植物的卵母细胞和颗粒细胞之间，这是发展和成熟所必需的 高质量的锻炼。已知河马信号传导调节组织尺寸。我们发表的结果建立了 河马信号传导的作用及其下游效应分子，YAP和TAZ作为颗粒的关键调节剂 细胞功能和生育能力。值得注意的是，我们的初步分析确定YAP/TAZ活动将被下调 患有IVF的晚期妇女（≥40岁）的产量低。我们假设那个yap TAZ策划了调节高质量伪装发展的关键转录事件。然而， 了解这些转录因子如何调节这些转录因子的实际机制存在差距 颗粒细胞功能，尤其是在接受IVF的衰老妇女中。该提议的总体目标是 了解YAP和TAZ介导的转录和染色质结构 高龄。使用初级患者颗粒样品的缺点是较低的样本数，这是主要的 障碍到进步。该提案的主要创新是同时剖面染色质和 同一患者样本中的转录调节，从而更深入地了解 颗粒细胞功能中的YAP和TAZ转录因子。我们的研究策略包括两个具体目标。 在特定的目标1中。我们将通过使用颗粒细胞中YAP和TAZ的转录靶标 Cut＆Run，这是一种可隔离特定蛋白质DNA复合物并区分的小焦点核酸酶技术 来自颗粒数量少的颗粒细胞的直接TF结合。我们的分析包括从IVF获得的颗粒细胞 反应较差（检索到6个卵母细胞）和良好反应（> 17个卵母细胞）的患者 高级产妇年龄（≥40岁）。我们还将使用ATAC-SEQ来确定染色质的可及性 这些患者样本。在AIM 2中，我们将在人颗粒细胞中调节YAP和TAZ，并定义它们 由此产生的转录组轮廓。这项研究中确定的基因将在功能上通过分析 颗粒细胞增殖，分化和感应。这些新颖的见解将有助于更好地理解 与年龄相关的生育能力下降以及河马信号通路在卵巢中影响YAP和TAZ的作用。