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.

项目总结 卵巢衰老大约在30岁左右开始，在接下来的10到15年里表现为不孕不育。一个 扩大对决定颗粒细胞功能的潜在分子机制的了解将有助于 我们对年龄和不孕不育的健康差距的理解。在过去的半个世纪里，第一批- 35岁及以上的高龄产妇的母亲的时间增加了五倍多。使用 更多的妇女进入劳动力大军，社会结构的变化，高等教育和更多地使用 在避孕方面，这一趋势可能会继续增加。有一种持续的双向交流 卵母细胞和卵泡颗粒细胞之间，这是卵泡发育和成熟所必需的 高质量的毛囊。众所周知，河马信号可以调节器官的大小。我们公布的结果确立了 河马信号及其下游效应分子YAP和TAZ作为颗粒细胞关键调节因子的作用 细胞功能和生育能力。值得注意的是，我们的初步分析发现YAP/TAZ活动在 高龄妇女(≥40岁)接受体外受精且卵子产量低。我们假设YAP 和TAZ协调关键的转录事件，调节高质量卵泡的发育。然而， 在了解这些转录因子如何调节的实际机制方面存在着知识空白 颗粒细胞功能，尤其是接受体外受精的老年妇女。这项提议的首要目标是 了解YAP和TAZ介导的卵巢颗粒细胞转录和染色质构筑 年事已高。使用原发患者颗粒样本的一个缺点是样本数量少，这是一个主要的 前进的路障。该方案的主要创新是能够同时对染色质和 在同一患者样本中的转录调节，从而提供了更深入的了解的作用 YAP和TAZ转录因子在颗粒细胞功能中的作用我们的研究策略包括两个具体目标。 在特定的目标1.我们将确定YAP和TAZ在颗粒细胞中的转录靶点 Cut&Run，一种微卡核酸酶技术，可以分离特定的蛋白质-DNA复合体并区分 从少量的颗粒细胞直接结合转铁蛋白。我们的分析包括从试管受精中获得的颗粒细胞 妇女中反应差(取回6个卵母细胞)和良好反应(取回17个卵母细胞)的患者 高龄产妇(≥40岁)。我们还将使用atac-seq来确定染色质在 这些病人样本。在目标2中，我们将在人颗粒细胞中调节YAP和TAZ，并确定它们的 由此产生的转录图谱。这项研究中确定的基因将通过分析 颗粒细胞的增殖、分化和衰老。这些新颖的见解将有助于更好地理解 与年龄相关的生育力下降以及河马信号通路效应器YAP和TAZ在卵巢中的作用。