IDENT OF DEAFNESS-CAUSING MUTATIONS USING PROTEOMICS-IDENTIFIED CANDIDATES

使用蛋白质组学鉴定的候选物鉴定导致耳聋的突变

• 批准号: 8365481
• 负责人: Peter Gordon Barr-Gillespie
• 金额: $ 2.87万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365481
• 关键词: Affect; Auditory; Auditory system; Biology; Chromosome Mapping; Complex; Couples; Disease; Epithelium; Equilibrium; Family; Funding; Genes; Genomics; Grant; Hair; Hair Cells; Hearing; Hereditary Disease; Human; Iowa; Knowledge; Labyrinth; Link; Membrane; Molecular; Molecular Genetics; Mus; Mutation; National Center for Research Resources; Organelles; Principal Investigator; Proteins; Proteome; Proteomics; Research; Research Infrastructure; Resources; Sensory Hair; Source; Techniques; United States National Institutes of Health; Universities; Work; cost; deafness; disease phenotype; hearing impairment; insight; member; mutant; next generation; novel; novel strategies; programs

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This new project couples proteomics (Gillespie), human genomics (Richard J. Smith, University of Iowa), and mouse molecular genetics (Ulrich M¿ller, Scripps) in an integrated program for identification and description of molecules essential for hearing and balance. The proteomics work will be done at OHSU/PNNL and is described here. When studying monogenic and complex genetic diseases, standard linkage mapping and/or association approaches to identify disease-relevant genes suffer from low throughput and lack of insight into function. Our approach provides high throughput in a contextual functional framework. We begin by noting that specific organelles are often implicated in disease. Focusing on the auditory system, we will first define the complete proteome (especially the membrane proteome) of the hair bundle, the mechanically sensitive organelle of sensory hair cells, which comprises <<1% of total protein in inner-ear epithelia. This work will be carried out at OHSU/PNNL by the Gillespie lab. Second, to link new genes to auditory disease, we will use next-generation sequencing to interrogate en masse the genes encoding bundle proteins in hundreds of affected families with uncharacterized forms of hearing loss. Sequencing will be carried out at Iowa by the Smith lab. Third, we will generate mouse lines carrying the orthologous human mutations and analyze the functional consequences of these mutations on bundle-protein networks. These mice will be generated by the M¿ller lab at Scripps, then mutant hair bundles will be examined using proteomic techniques by the Gillespie lab at OHSU/PNNL. This project is iterative, so novel members of disrupted networks will be screened in human families as well. This composite approach provides an understanding of a disease phenotype at the molecular level, knowledge requisite to developing novel approaches to disease treatment.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 这一新项目将蛋白质组学(Gillesbie)、人类基因组学(Richard J.Smith，爱荷华大学)和小鼠分子遗传学(Ulrich M？ler，Scripps)结合在一个综合程序中，用于识别和描述对听力和平衡至关重要的分子。蛋白质组学工作将在OHSU/PNNL完成，并在这里进行描述。在研究单基因和复杂遗传病时，识别疾病相关基因的标准连锁图谱和/或关联方法存在吞吐量低和缺乏对功能的洞察。我们的方法在上下文功能框架中提供高吞吐量。我们首先注意到，特定的细胞器通常与疾病有关。以听觉系统为中心，我们将首先定义毛束的完整蛋白质组(特别是膜蛋白质组)，毛束是感觉性毛细胞的机械敏感细胞器，占内耳上皮细胞总蛋白质的1%。这项工作将由吉莱斯皮实验室在OHSU/PNNL进行。其次，为了将新基因与听力疾病联系起来，我们将使用下一代测序技术对数百个患有不明形式听力损失的受影响家庭的编码束蛋白的基因进行集体询问。测序将由史密斯实验室在爱荷华州进行。第三，我们将产生携带人类同源突变的小鼠系，并分析这些突变对束-蛋白质网络的功能后果。这些小鼠将由斯克里普斯的M？ler实验室培育出来，然后OHSU/PNNL的吉莱斯皮实验室将使用蛋白质组学技术对突变的发束进行检测。这个项目是迭代的，所以被破坏的网络的新成员也将在人类家庭中进行筛选。这种综合方法在分子水平上提供了对疾病表型的理解，这是开发新的疾病治疗方法所必需的知识。