A model system for pathological transitions of RNA-binding protein aggregates

项目摘要/摘要 与年龄相关的神经退行性疾病是一类无法治愈的疾病，导致进展性 神经细胞变性。最近的研究表明，RNA结合蛋白在年龄中的作用- 相关神经退行性疾病：因为许多RNA结合蛋白具有聚集倾向 本质上无序的区域，它们聚集在不同类型的病理性包裹体中 神经退行性疾病和一些RNA结合蛋白的突变与 神经退行性疾病。许多与神经退行性疾病有关的RNA结合蛋白 在应激条件下，它们也会浓缩成mRNP颗粒。这是一种 应激过程中的MRNP颗粒对细胞是有益的，因为这些颗粒的破坏使细胞对 各种压力源。我们的假设是mRNP颗粒最初形成为无毒的、潜在的 在衰老过程中的有益结构，但它们经历了一种病理转变，从而导致 致细胞死亡。这项提议试图建立一个模型系统，用于这种病理转变，使用 酵母菌联合微流控跟踪mRNP颗粒的形成和转化 衰老。为了实现这一点，我们将(目标1)建立在初步数据的基础上，探索沙门氏菌的致病性 用微流控和荧光显微镜跟踪单个酵母的AGE诱导的mRNP大颗粒 细胞的整个生命周期。其次，我们将(目标2)确定这种病理的标志物 通过测量AGE诱导的物质状态和大分子组成的变化来实现转变 MRNP颗粒。从长远来看，这些标记将有助于阐明这一机制的细节 过渡。总而言之，这个系统将建立一个新的范式，以了解关于如何 MRNP颗粒可以从无毒的、潜在有益的结构转变为病理性包裹体 会导致细胞退化。因为许多基因和途径从酵母到 哺乳动物，包括影响衰老的途径，这项研究可能指出新的靶点，以帮助 改善人类神经退行性疾病。

Project Summary/Abstract Age-related neurodegenerative diseases are a class of incurable diseases that result in the progressive degeneration of neuronal cells. Recent research has pointed to a role for RNA-binding proteins in age- related neurodegenerative diseases: as many RNA-binding proteins have aggregation-prone intrinsically disordered regions, they accumulate in pathological inclusions of various neurodegenerative diseases, and mutations in a number of RNA-binding proteins have been linked to neurodegenerative diseases. For many RNA-binding proteins linked to neurodegenerative diseases they also become concentrated into mRNP granules during stressful conditions. This formation of mRNP granules during stress is beneficial to cells, as disruption of these granules sensitizes cells to a variety of stressors. Our hypothesis is that mRNP granules initially form as non-toxic, potentially beneficial structures during the aging process, but that they undergo a pathological transition that leads to cell death. This proposal seeks to establish a model system for this pathological transition using yeast in combination with microfluidics to follow the formation and transition of mRNP granules during aging. To accomplish this, we will (Aim 1) build on our preliminary data exploring the pathogenicity of large age-induced mRNP granules using microfluidics and fluorescent microscopy to follow single yeast cells across their entire lifespan. Secondly, we will (Aim 2) identify markers for this pathological transition by measuring changes in the material state and macromolecular composition of age-induced mRNP granules. In the long term these markers will help to elucidate the mechanistic details of this transition. Together this system will establish a new paradigm to understand basic details on how mRNP granules can transition from non-toxic, potentially beneficial structures to pathological inclusions that drive the degeneration of cells. As many genes and pathways are conserved from yeast to mammals, including pathways that affect aging, this research may point to new targets to help ameliorate neurodegenerative diseases in humans.