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

项目总结/摘要 选择性剪接是一种基本的基因调控机制，它允许细胞显着多样化其功能。 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 调节剪接的限制性内切酶（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，尽管出现了这种模式，很少有剪接因子被研究， 关于衰老和与衰老有关的疾病。这个项目的长期目标是开始阐明如何 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Caper将被用作模型。Caper是必需的 果蝇感觉神经元的发育和维持，以及成年运动行为。我们 初步数据显示，跳跃功能障碍导致寿命缩短和运动行为下降 随着年龄的增长而恶化此外，这些老化表型显示出强烈的性别偏见，其中男性是 比女性更受影响。这种性别偏见经常出现在与衰老有关的疾病中，但潜在的 这种性别偏见的机制尚不清楚。尽管我们对人类的功能知之甚少 刺山柑直系同源物，人刺山柑直系同源物RBM 39，与癌症进展有关， 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）Caper 与女性相比，在男性中差异剪接;（3）Caper调节衰老过程中的性别特异性剪接。 使用高度易处理的模型，果蝇，我们将利用免疫荧光来检查各种标志物， 成年果蝇的脑解剖、细胞凋亡和线粒体标志物，以确定是否存在跳跃功能障碍 导致神经退化我们将使用差异基因表达和剪接形式分析来确定 在衰老的雄性和雌性果蝇中，雀跃本身是否有差异剪接，以及雀跃是否调节了差异剪接。 其他基因的拼接在老化的雄性和雌性果蝇中。最后，我们将进行试点遗传筛查， 鉴定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 该基金会将开展一项更全面的研究计划，旨在利用刺山柑作为衰老和性的模型， 对衰老相关疾病的偏见由于异常的选择性剪接已经成为一个共同的主题， 各种衰老疾病，从这项研究中获得的知识具有广泛的意义， 治疗这些疾病。

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.