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: 10537634
• 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/10537634
• 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; Genetic Translation; 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; 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; Testing; Time; Tissues; Transcript; Translation Initiation; Translations; Work; age related; base; 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; oocyte quality; paralogous gene; proteostasis; reproductive senescence; reproductive success; response; therapy design/development; tool; translation factor

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。在储存的女性生殖系统中尤其如此 卵母细胞已直接与流产和先天缺陷的发生率增加有关。我们的 长期目标是确定并表征导致生殖衰老的因素。在 哺乳动物，鸡蛋可以存储数月，数年或几十年，进行生殖衰老的分析 缓慢而实验困难。在这里，我们试图基于以前建立果蝇的努力 卵巢是一种强大的系统，可以研究生殖衰老。有趣的是，鸡蛋的下降 从苍蝇到人类，质量与跨物种的mRNA平移水平较低相关。 尽管存在这种常见的缺陷，但我们对此的机制知之甚少 储存鸡蛋内的mRNA翻译能力的降低。在这里，我们建议使用最新技术 基因操纵和生化分析系统地表征机械的方式 mRNA翻译随着产妇年龄的变化和果蝇储存期间所需的所需。 此外，我们寻求基因测试是否操纵核糖体水平和翻译 起始/伸长率延长了储存的鸡蛋的质量。我们已经建立了一个运营 管道进行该提案中概述的所有实验的管道，并寻求利用 我们小组采用的许多创新工具和技术。在AIM 1下，我们 将使用互补的分子和生化方法全面地表征 蛋白质合成和核糖体水平在衰老女性卵巢中发生变化的程度 鸡蛋存储了两个星期。我们还将使用基于生化和创新的记者测定法 评估翻译保真度是否随着年龄的增长而下降。在AIM 2中，我们将测试 提高或降低核糖体水平以及翻译起始和伸长率可改善 储存鸡蛋的质量。在AIM 3下，我们将表征核糖体氧合酶NO66如何影响 鸡蛋质量。我们认为，在衰老过程中对体内卵母细胞的全面分析将 提供关键的见解，说明为什么卵的质量随着年龄的增长而下降，并将揭示新的分子靶标 为了开发旨在改善和扩展繁殖的疗法。鉴于我们专注于 角色核糖体在此过程中发挥作用，我们认为我们的工作将广泛影响组织的研究 衰老生物的稳态和再生。