Quantitative Proteomic Approach to Identify the Mechanism of Alzheimer's Disease

描述(由申请人提供)： 哺乳动物的大脑是一个极其复杂的器官，在驻留细胞类型、解剖组织、神经元回路、细胞间通讯模式和蛋白质表达调控方面都是如此。此外，衰老和阿尔茨海默病(AD)对大脑的影响对研究人员来说是一个巨大的挑战。以前的基于质谱学的蛋白质组学技术最擅长于识别单一细胞类型的浓缩蛋白质的蛋白质-蛋白质相互作用或翻译后修饰。哺乳动物体内同位素标记和高分辨率质谱仪的最新发展为确定组织中数千种蛋白质的相对蛋白质表达水平提供了机会。我们的目标是通过定量质谱学计算在阿尔茨海默病进展和哺乳动物大脑规则衰老过程中蛋白质表达的变化。这个项目将产生一个前所未有的大规模蛋白质表达解剖清单，可以在未来几年内挖掘出来。这些定量的蛋白质组学研究将作为一个假设生成机器，可能会发现关于衰老和AD对大脑的影响的新的和意想不到的数据。那些处于AD晚期的人变得卧床不起，每天24小时依赖护理，这导致仅在美国每年就有1480亿美元的成本。衰老和阿尔茨海默病对大脑生理学的影响历来是以候选人为基础的研究方法。虽然这些候选方法对我们理解衰老和阿尔茨海默病有很大贡献，但它们的范围有限。为了扩大我们对这些过程的知识基础，并最终开发出治疗AD的有效疗法，我们将计算数千种蛋白质的表达水平。表达受干扰的蛋白质将作为识别与AD病理和衰老相关的途径的灯塔。扰动途径的确定(S)将反过来为旨在破译AD和衰老的分子基础的深入分析提供肥沃的土壤。更广泛地说，这个项目将提供在衰老和AD期间发生的蛋白质表达的大脑图谱。AD是对世界老龄化人口的重大威胁：AD被认为是世界上最常见的神经退行性疾病，影响着500多万老年美国人，并对公共健康构成越来越大的威胁。AD剥夺了个人的推理、使用语言和回忆记忆的能力，从而削弱了他们的能力，导致了巨大的照顾负担。目前，阿尔茨海默病还没有治愈或明确的治疗方法，它仍然是导致痴呆症的主要原因。AD按发病年龄分层。阿尔茨海默病的两种形式，早发性和晚发性，都有遗传联系。识别与AD病理有关的新基因和/或蛋白质可能为开发有效的AD治疗方法提供关键的第一步。 公共卫生相关性： 阿尔茨海默病(AD)使65岁或65岁以上的人的医疗费用增加了两倍，目前估计有530万美国人受到影响。在这里，我们建议完成第一个大规模的蛋白质在衰老和AD进展过程中表达的解剖学清单。这些研究应该以前所未有的水平进行分析，并通过这样做，揭示与成功治疗这种毁灭性疾病相关的新途径。

DESCRIPTION (provided by applicant): The mammalian brain is an immensely complex organ in terms of diversity of resident cell types, anatomical organization, neuronal circuitry, modes of intercellular communication, and the regulation of protein expression. Further the effect aging and Alzheimer's disease (AD) has on the brain represents a colossal challenge for researchers. Previous mass spectrometry based proteomic technologies have been most adept at identifying protein-protein interactions or post-translational modifications of enriched proteins from a single cell type. The recent development of in vivo isotopic labeling of mammals and high resolution mass spectrometers has afforded the opportunity to determine the relative protein expression level of thousands of proteins from tissue. We aim to calculate changes in protein expression during Alzheimer's disease progression and regular aging in the mammalian brain by quantitative mass spectrometry. This project will yield a large-scale anatomic inventory of protein expression on an unprecedented level which can be mined for years to come. These quantitative proteomic studies will serve as a hypothesis-generating machine that will likely uncover new and unexpected data on the effect aging and AD have on the brain. Those in the advanced stages of AD become bed-bound and reliant on care 24/7 which in total results in 148 billion dollars in annual costs in the US alone. The effect of aging and AD on brain physiology has been historically investigated in candidate-based approaches. While these candidate approaches have significantly contributed to our understanding of aging and AD, they have been limited by their restricted scope. To broaden our knowledge base of these processes and eventually develop effective therapeutics for the treatment of AD we will calculate the expression level for thousands of proteins. Proteins with perturbed expression will serve as beacons for the identification of pathways relevant to AD pathology and aging. Determination of perturbed pathway(s) will in turn serve as fertile ground for in-depth analysis aimed at deciphering the molecular basis of AD and aging. More generally, this project will deliver a brain atlas of protein expression that occurs during aging and AD. AD represents a significant threat to the aging world population: AD is considered the world's most common neurodegenerative disease, affecting over 5 million aging Americans, and is a rising threat to public health. AD debilitates individuals by stripping them of the ability to reason, use language, and recall memories, resulting in a tremendous caretaking burden. Currently there is no cure or definitive treatment for AD and it remains the leading cause of dementia. AD is stratified by the age at which pathological onset occurs. The two forms of AD, early-onset and late-onset, both have genetic links. Identification of new genes and/or proteins that contribute to AD pathology could provide a critical first step for the development of effective AD treatments. PUBLIC HEALTH RELEVANCE: Alzheimer's disease (AD) triples health care costs for those aged 65 or older and currently affects an estimated 5.3 million Americans. Here, we propose to complete the first large-scale anatomic inventory of protein expression during aging and AD progression. These studies should yield analysis at an unprecedented level and, through doing so, reveal novel pathways relevant to the successful treatment of this devastating disease.