Proteome Biology of Noise Induced Hearing Loss

描述（由申请人提供）：听力障碍是最常见的感觉障碍之一，全世界有2.5亿人患有中度至重度听力损失（38），由于语言交流的核心作用，大大降低了生活质量。在经济规模上，听力损失的负面影响总和大于多发性硬化症、脊髓损伤、中风、癫痫、帕金森病和亨廷顿病的总和，影响的人数是后者的4倍（68）。在确定的听力损失病因中，最常见的导致因素是过度噪音，数百万人在工作时暴露在危险的大噪音中。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，通过识别重要的蛋白质和途径负责听力损失。具体而言，我们开发了一个定量蛋白质组学分析平台，以探测过量噪声对耳蜗蛋白质组的影响。我们的初步数据表明，这种方法可以准确地测量来自一只老鼠的数千种蛋白质，并且已经揭示了噪音暴露后蛋白质的显着扰动。我们还在分离Corti器官方面取得了进展，以确保准确测量头发和邻近支持细胞以及耳蜗神经突触中含量低但可能改变的蛋白质。更具体地说，我们将定量分析暴露于多种噪音水平的小鼠的内耳提取物。通过这些比较暴露，我们将区分具有受过量噪声影响的特征的蛋白质。这些蛋白质组学实验的候选物将通过生物信息学工具进行探索，并通过传统的基于抗体的方法进行验证。下一步，我们将开发生化方法，以确保稀有低丰度蛋白质的准确测量。我们还将测试蛋白质-蛋白质相互作用是否在表达水平没有显著变化的情况下被破坏。最后，我们将使用已知的生物活性分子来保护NIHL，并重复蛋白质组学分析来研究这些药物有效的机制。特别是，我们认为对内耳蛋白质组的全面了解将加速听力损伤的更大研究领域。总之，我们在此建议通过应用定量蛋白质组学工具来识别和研究NIHL中的分子缺陷，该工具可以在一次分析中同时敏感地研究数千种蛋白质。我们相信这一建议代表了定量蛋白质组学在研究中的首次应用

DESCRIPTION (provided by applicant): 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 changs 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.