Proteome Biology of Noise Induced Hearing Loss

描述(申请人提供)：听力障碍是最常见的感官障碍之一，全球有2.5亿人患有中到重度听力损失(38人)，由于语言交流的核心作用，显著降低了生活质量。在经济规模上，听力损失的总体负面影响大于多发性硬化症、脊髓损伤、中风、癫痫、帕金森氏症和亨廷顿病的总和，影响人数是后者的4倍(68人)。在已定义的听力损失病因中，最常见的原因是过度噪音，数百万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结果将有助于通过识别导致听力损失的重要蛋白质和途径来减少噪音导致的听力损失。具体地说，我们开发了一个定量蛋白质组分析平台来探索过量噪声对耳蜗蛋白质组的影响。我们的初步数据显示，这种方法可以准确地测量一只小鼠的数千种蛋白质，并且已经揭示了噪音暴露后明显扰动的蛋白质。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低丰度但可能发生变化的蛋白质。更具体地说，我们将对暴露在多种水平噪音中的小鼠的内耳提取物进行定量分析。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。这些蛋白质组实验的候选者将用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。接下来，我们将开发生物化学方法，以确保稀有低丰度蛋白的准确测量。我们还将测试蛋白质之间的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，我们将使用已知的保护NIHL的生物活性分子，并重复蛋白质组分析，以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，我们建议通过应用定量蛋白质组学工具来识别和研究NIHL的分子缺陷，这种工具可以在一次分析中同时灵敏地研究数千种蛋白质。我们相信这一提议代表了定量蛋白质组学在研究中的首次应用。 可能拥有所需的分析力量，以启动向最终治疗和预防NIHL的有效疗法的发展。

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.