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.

项目摘要/摘要 替代剪接是一种基本基因调节机制，可使细胞显着多样化它们 蛋白质产品。剪接缺陷，通常是由破坏RNA结合蛋白功能的突变引起的 （RBP）调节剪接，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，尽管有这种新兴模式，但很少研究剪接因素 关于衰老和与衰老有关的疾病。该项目的长期目标是开始阐明 由高度保守的RBP功能障碍引起的异常剪接，导致异常衰老 表型。为此，高度保守的剪接因子帽帽将用作模型。需要刺山柑 为了发展和维持果蝇感觉神经元以及成人运动行为。我们的 初步数据表明，刺山裁判功能障碍会导致寿命缩短和运动行为下降 随着年龄的增长而加剧。此外，这些老化表型显示出强烈的性偏见，男性是 比女性更受影响。这种性偏见经常在与衰老有关的疾病中看到 这种性偏见的机制仍然未知。虽然对人的功能知之甚少 开普敦直系同源物，人雀跃直系同源物rbm39，与癌症进展有关，并表达 在整个神经系统中。本应用程序中提出的研究将测试以下 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，男性的剪接是差异的。 （3）CAPER在衰老过程中调节性别特定的剪接。 使用果蝇高度可行的模型，我们将利用免疫荧光检查 脑部解剖学，凋亡和线粒体标记随着时间的流逝，成人果蝇，以确定刺囊功能障碍是否功能障碍 导致神经变性。我们将使用差异基因表达和剪接分析来确定 刺山柑本身是否在衰老的雄性与雌性苍蝇中差异化，并且云母是否调节差异 衰老与雌性苍蝇中其他基因的剪接。最后，我们将执行一个试验遗传屏幕 识别与刺山柑相互作用以调节衰老表型的基因。这项研究的结果将建立 更全面的研究计划的基础，旨在将开普敦用作衰老和性别的模型 与衰老有关的疾病的偏见。由于异常替代拼接已成为一个共同的主题 各种衰老疾病，这项研究所获得的知识对理解和 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.