Proteome Biology of Noise Induced Hearing Loss

项目总结 听力障碍是最常见的感官障碍之一，2.5亿人 全世界有中到重度听力损失(38人)，并显著降低生活质量 由于言语交流的中心作用。在经济规模上，总的负面影响 听力损失的影响比多发性硬化症、脊髓损伤、中风、 癫痫、帕金森氏症和亨廷顿病加在一起，影响的人数是 (68)。在已确定的听力损失病因中，最常见的致病因素是过度 噪音，数以百万计的人在工作中暴露在危险的巨大噪音中。我们希望我们的 研究发现将有助于减少噪音引起的听力损失，因为 导致听力损失的蛋白质和通路。具体地说，我们开发了一种 探讨过量噪声对耳蜗组织影响的蛋白质组学定量分析平台 蛋白质组。我们的初步数据表明，这种方法可以准确地测量数千个 来自一只小鼠的蛋白质，并已经发现蛋白质在 噪音暴露。我们在分离科尔蒂器官方面也取得了进展，以确保 在头发和邻近的支持细胞中测量低丰度但可能发生变化的蛋白质， 以及在耳蜗神经突触中。更具体地说，我们将对内耳进行量化分析 暴露在不同水平噪音下的小鼠的提取物。通过这些可比较的暴露， 我们将区分具有受过多噪音影响的特征的蛋白质。 这些蛋白质组学实验的候选者将用生物信息学来探索 工具，并通过传统的基于抗体的方法进行验证。接下来，我们将发展生物化学 方法确保稀有低丰度蛋白的准确测定。我们还将测试 如果蛋白质之间的相互作用被破坏，而表达水平没有显著变化。 最后，我们将使用已知的生物活性分子来预防NIHL，并重复蛋白质组学 分析以研究这些药物的有效机制。特别是，我们 认为全面了解内耳蛋白质组会加速更大 听力损伤的研究领域。总而言之，我们建议在这里确定和调查 应用定量蛋白质组学工具研究NIHL中的分子缺陷 在一次分析中灵敏地研究数千种蛋白质。我们相信这项建议 代表了定量蛋白质组学首次应用于NIHL的研究和 可以拥有所需的分析力量来推动向有效发展 最终治疗和预防NIHL。

PROJECT SUMMARY Hearing impairment is one of the most common sensory disabilities, 250 million people worldwide have moderate to severe hearing loss (38), and significantly reduces quality of life due to the central role of verbal communication. On an economic scale, the total negative impact of hearing loss is greater than that of multiple sclerosis, spinal cord injury, stroke, epilepsy, Parkinson's and Huntington's disease combined and effects 4 times as many people (68). The most common causative factor among the defined hearing loss etiologies is excessive noise, and millions of people are exposed to dangerously loud noise at work. We hope that our research findings will aid in the reduction of noise-induced hearing loss by identifying significant proteins and pathways responsible for hearing loss. Specifically, we have developed a quantitative proteomic analysis platform to probe the effect of excess noise on the cochlear proteome. Our preliminary data shows this approach can accurately measure thousands of proteins from a single mouse and has already revealed proteins significantly perturbed after noise exposure. We have also made progress isolating the organ of Corti to ensure the accurate measurement of low abundant but potentially altered proteins in hair and adjacent support cells, and in cochlear nerve synapses. More specifically, we will quantitatively analyze inner ear extracts from mice exposed to multiple levels of noise. Through these comparative exposures, we will differentiate proteins with characteristics that are impacted by excess noise. The candidates from these proteomic experiments will be explored with bioinformatic tools and validated by traditional antibody based approaches. Next we will develop biochemical methods to ensure the accurate measurement of rare low abundance proteins. We will also test if protein–protein interactions are disrupted without significant changes in expression levels. Finally we will use bioactive molecules known to protect from NIHL and repeat the proteomic analysis to investigate the mechanisms by which these drugs are effective. In particular, we think that a comprehensive understanding of the inner ear proteome will accelerate the greater research field of hearing injury. In summary, we propose here to identify and investigate molecular defects in NIHL by applying quantitative proteomic tools that can simultaneously and sensitively investigate thousands of proteins in a single analysis. We believe this proposal represents the first ever application of quantitative proteomics to the investigation of NIHL and may hold the required analytical strength to kick start the development towards effective therapeutics to eventually treat and prevent NIHL.