Characterization of how mRNA translation influences reproductive aging

mRNA 翻译如何影响生殖衰老的表征

• 批准号: 10665757
• 负责人: Michael Buszczak
• 金额: $ 33.62万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665757
• 关键词: Adopted; Affect; Age; Aging; Biochemical; Biogenesis; Biological Assay; Cell physiology; Cells; Cellular biology; Complex; Congenital Abnormality; Data; Defect; Disease; Drosophila genus; Embryonic Development; Exhibits; Female; Gene Modified; Genetic; Genetic Models; Genetic Transcription; Germ Cells; Goals; Growth; Health; Homeostasis; Human; Hydroxylation; Incidence; Intervention; Knock-out; Length; Link; Mammals; Maternal Age; Messenger RNA; Modeling; Modification; Molecular; Molecular Biology; Molecular Target; Natural regeneration; Oocytes; Oogenesis; Organelles; Organism; Ovarian aging; Ovary; Oxidation-Reduction; Oxygenases; Phenotype; Physiological; Play; Post-Translational Protein Processing; Process; Production; Protein Biosynthesis; Proteins; Publishing; Reagent; Reporter; Reproduction; Reproductive Health; Reproductive Process; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Risk Factors; Role; Signal Transduction; Spontaneous abortion; Study models; System; Techniques; Technology; Testing; Time; Tissues; Transcript; Translation Initiation; Translations; Work; age related; biochemical model; egg; experience; experimental study; female reproductive system; fly; gain of function; genetic manipulation; genomic locus; improved; in vivo; innovation; insight; loss of function; mRNA Translation; oocyte quality; paralogous gene; proteostasis; reproductive senescence; reproductive success; response; therapy design/development; tool; translation factor; translational impact

Summary Aging represents a major risk factor for a broad range of diseases and declines in tissue homeostasis and function. This is particularly true in the female reproductive system where the aging of stored oocytes has been directly linked with an increased incidence of miscarriages and birth defects. Our long-term goal is to identify and characterize the factors that contribute to reproductive aging. In mammals, eggs can be stored months, years, or decades, making the analysis of reproductive aging slow and experimentally difficult. Here, we seek to build upon previous efforts to establish the Drosophila ovary as a powerful system with which to study reproductive aging. Interestingly, the decline in egg quality has been correlated with lower levels of mRNA translation across species, from flies to humans. Despite this common defect, we know surprisingly little about the mechanisms responsible for this reduction of mRNA translation capacity within stored eggs. Here, we propose to use state of the art genetic manipulation and biochemical analysis to systematically characterize how the machinery required for mRNA translation changes with maternal age and during egg storage in Drosophila. Moreover, we seek to genetically test whether manipulating ribosome levels and translation initiation/elongation rates prolongs the quality of stored eggs. We have established an operational pipeline for conducting all the experiments outlined under this proposal and seek to take advantage of a number of innovative tools and techniques that have been adopted by our group. Under Aim 1, we will use complementary molecular and biochemical approaches to comprehensively characterize the extent to which protein synthesis and ribosome levels changes in the ovaries of aging females and in eggs stored over two weeks. We will also use biochemical and innovative reporter based assays to evaluate whether translation fidelity declines with age. In aim 2, we will test the extent to which increasing or decreasing ribosome levels and translation initiation and elongation rates improves the quality of stored eggs. Under aim 3, we will characterize how the ribosome oxygenase NO66 influences egg quality. We believe this comprehensive analysis of in vivo oocytes during the course of aging will provide key insights into why the quality of eggs declines with age and will reveal new molecular targets for the development of therapies designed to improve and extend reproduction. Given our focus on the role ribosomes play in this process, we believe our work will broadly impact the study of tissue homeostasis and regeneration in aging organisms.

概括 衰老是多种疾病和组织稳态下降的主要危险因素 和功能。这在女性生殖系统中尤其如此，其中存储的老化 卵母细胞与流产和出生缺陷发生率增加直接相关。我们的 长期目标是确定和描述导致生殖衰老的因素。在 哺乳动物的卵可以保存数月、数年或数十年，从而进行生殖衰老分析 缓慢且实验困难。在这里，我们寻求在之前建立果蝇研究中心的努力基础上再接再厉。 卵巢作为研究生殖衰老的强大系统。有趣的是，鸡蛋的减少 从果蝇到人类，跨物种的质量与较低水平的 mRNA 翻译相关。 尽管存在这种常见缺陷，但令人惊讶的是我们对造成这种情况的机制知之甚少 储存鸡蛋内 mRNA 翻译能力降低。在这里，我们建议使用最先进的技术 基因操作和生化分析系统地描述了机器如何 mRNA 翻译所需的基因会随着母体年龄和果蝇卵储存期间的变化而变化。 此外，我们试图从基因角度测试是否操纵核糖体水平和翻译 起始/延伸率可延长储存鸡蛋的质量。我们已经建立了一个可操作的 进行本提案中概述的所有实验的管道，并寻求利用 我们小组采用了许多创新工具和技术。在目标 1 下，我们 将使用互补的分子和生化方法来全面表征 老年女性卵巢中蛋白质合成和核糖体水平变化的程度 鸡蛋储存超过两周。我们还将使用生化和创新的基于报告的测定法 评估翻译保真度是否随着年龄的增长而下降。在目标 2 中，我们将测试 增加或减少核糖体水平以及翻译起始和延伸率可改善 储存鸡蛋的质量。在目标 3 下，我们将描述核糖体加氧酶 NO66 如何影响 鸡蛋质量。我们相信，这种对衰老过程中体内卵母细胞的全面分析将 提供关于卵子质量为何随着年龄增长而下降的重要见解，并将揭示新的分子靶点 开发旨在改善和延长生殖的疗法。鉴于我们的重点是 核糖体在此过程中发挥的作用，我们相信我们的工作将广泛影响组织的研究 衰老有机体的稳态和再生。