A pilot study to investigate the contribution of alternative splicing to sex biases of aging related phenotypes

一项初步研究，调查选择性剪接对衰老相关表型性别偏差的影响

• 批准号: 10176350
• 负责人: Jeremy Matthew Bono
• 金额: $ 7.2万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176350
• 关键词: Adult; Affect; Afferent Neurons; Age; Aging; Alternative Splicing; Amyotrophic Lateral Sclerosis; Anatomy; Animals; Apoptosis; Behavior; Behavioral Assay; Brain; Capers; Cells; Collection; Data; Data Set; Defect; Development; Disease; Distant; Drosophila genus; Female; Foundations; Functional disorder; Gene Expression Regulation; Genes; Genetic Screening; Goals; Grooming; Human; Immunofluorescence Immunologic; In Situ Nick-End Labeling; Knowledge; Link; Locomotion; Longevity; Maintenance; Malignant Neoplasms; Mammals; Mediating; Mitochondria; Modeling; Molecular; Molecular Genetics; Morphology; Mutation; Nerve Degeneration; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neurologic; Neurons; Organ; Orthologous Gene; Outcome; Outcome Study; Pattern; Phenotype; Pilot Projects; Population; Protein Isoforms; Proteins; Proteome; Public Health; RNA; RNA Splicing; RNA-Binding Proteins; Reagent; Regulation; Regulator Genes; Research; Research Proposals; Role; Sensory; Sex Bias; Site; Stains; Testing; Time; Tissue-Specific Gene Expression; behavioral phenotyping; differential expression; fly; gene function; genetic manipulation; insight; knock-down; male; mature animal; mutant; neuron development; normal aging; novel; programs; sex; transcriptomics; tumor progression

Project Summary/Abstract Alternative splicing is a fundamental gene regulatory mechanism that allows cells to significantly diversify their protein products. Splicing defects, often caused by mutations that disrupt the function of RNA-binding proteins (RBPs), which regulate splicing, have increasingly been implicated in aging-related disorders including cancer and neurodegenerative disease. Yet, despite this emerging pattern, few splicing factors have been studied with regard to aging and aging-related disease. The long-term goal of this project is to begin to elucidate how aberrant splicing that results from the dysfunction of a highly conserved RBP, leads to abnormal aging phenotypes. To this end, the highly conserved splicing factor Caper will be used as a model. Caper is required for the development and maintenance of Drosophila sensory neurons, and for adult locomotor behavior. Our preliminary data show that caper dysfunction leads to a shortened lifespan and a decline in locomotor behavior that is exacerbated with age. Moreover, these aging phenotypes show a strong sex bias, where males are more affected than females. Such sex biases are often seen in aging-related diseases, but the underlying mechanism for such sex biases remains unknown. Though little is known about the function of the human caper ortholog, the human caper ortholog, RBM39, is implicated in cancer progression and is expressed throughout the nervous system. The research proposed within this application will test the following hypotheses: (1) Caper regulates the maintenance of neurons in the adult brain and nervous system; (2) caper is differentially spliced in males compared to females; (3) Caper regulates sex-specific splicing during aging. Using the highly tractable model, Drosophila, we will utilize immunofluorescence to examine various markers of brain anatomy, apoptosis and mitochondrial markers over time in adult flies to determine if caper dysfunction results in neurodegeneration. We will use differential gene expression and spliceform analyses to determine whether caper itself is differentially spliced in aging male versus female flies, and if caper regulates differential splicing of other genes in aging male versus female flies. Finally, we will perform a pilot genetic screen to identify genes that interact with caper to regulate aging phenotypes. The outcomes of this study will establish the foundation for a more comprehensive research program aimed at using caper as a model for aging and sex biases in aging-related disease. Since aberrant alternative splicing has emerged as a common theme in various aging disorders, the knowledge gained from this study has broad implications for understanding and treating such disorders.

项目概要/摘要 选择性剪接是一种基本的基因调控机制，可以使细胞显着多样化 蛋白质产品。剪接缺陷，通常由破坏 RNA 结合蛋白功能的突变引起 调节剪接的 RBP（RBP）越来越多地与包括癌症在内的衰老相关疾病有关 和神经退行性疾病。然而，尽管存在这种新兴模式，但很少有研究人员对剪接因子进行了研究。 关于衰老和与衰老相关的疾病。该项目的长期目标是开始阐明如何 高度保守的 RBP 功能障碍导致异常剪接，导致异常衰老 表型。为此，高度保守的剪接因子Caper将被用作模型。刺山柑是必需的 用于果蝇感觉神经元的发育和维持，以及成年运动行为。我们的 初步数据显示雀跃功能障碍会导致寿命缩短和运动行为下降 随着年龄的增长，这种情况会加剧。此外，这些衰老表型表现出强烈的性别偏见，其中男性是 比女性受影响更大。这种性别偏见常见于与衰老相关的疾病，但其根本原因是 这种性别偏见的机制仍然未知。尽管人们对人体的功能知之甚少 caper 直向同源物，人类 caper 直向同源物 RBM39，与癌症进展有关并表达 整个神经系统。本申请中提出的研究将测试以下内容 假设：（1）Caper 调节成人大脑和神经系统中神经元的维持； (2) 刺山柑 与女性相比，男性的剪接存在差异； (3) Caper在衰老过程中调节性别特异性剪接。 使用高度易处理的模型果蝇，我们将利用免疫荧光来检查果蝇的各种标记物 成年果蝇的大脑解剖结构、细胞凋亡和线粒体标记随时间的变化，以确定雀跃功能障碍是否存在 导致神经退行性变。我们将使用差异基因表达和剪接形式分析来确定 刺山柑本身在衰老的雄性果蝇和雌性果蝇中是否存在差异剪接，以及刺山柑是否调节差异剪接 衰老雄性果蝇与雌性果蝇中其他基因的剪接。最后，我们将进行试点基因筛查 识别与刺山柑相互作用以调节衰老表型的基因。这项研究的结果将确定 为更全面的研究计划奠定了基础，旨在利用刺山柑作为衰老和性的模型 与衰老相关的疾病存在偏见。由于异常选择性剪接已成为一个常见主题 各种衰老疾病，从这项研究中获得的知识对于理解和 治疗此类疾病。

项目成果

Roles for the RNA-Binding Protein Caper in Reproductive Output in Drosophila melanogaster.

• DOI: 10.3390/jdb11010002
• 发表时间: 2022-12-23
• 期刊: JOURNAL OF DEVELOPMENTAL BIOLOGY
• 影响因子: 2.7
• 作者: Tixtha, Erika J.;Super, Meg K.;Titus, M. Brandon;Bono, Jeremy M.;Olesnicky, Eugenia C.
• 通讯作者: Olesnicky, Eugenia C.