Alzheimer's Disease Characterization via a Novel Native Mass Spectrometry Platform

摘要 目前的医疗环境还不具备应对阿尔茨海默氏症的能力，预计 2010至2050年间，美国人口增加了两倍，医疗保健成本高达数千亿美元 每年。为了更好地了解和诊断这种疾病，人们的注意力已经转移到生物标志物上。这个 低聚形式的淀粉样β蛋白-可以被金属离子稳定-已经被标记为最多的 阿尔茨海默氏症背后的破坏性驱动力。在这种疾病中，质谱学是一种突出的力量 特征，但由于显著的非共价景观的淀粉样β蛋白，质量 光谱分析必须以一种保留内源性分子背景的方式来应用，以获得最大的效果。 自然自上而下的质谱学改进了从标准方法获得的信息 通过保持分子间和分子内的非共价相互作用。但是，该方法在 由于大型分析物的数据采集需要手动操作，因此可实现高吞吐量。另一个障碍是 通过质谱学对非共价蛋白质组件的表征是缺乏用于 不稳定改性的本地化(例如，金属)。拟议的工作将通过以下方式解决第一个障碍 自动重新配置质谱仪的软件，以增强每个分析物的传输 人物刻画。优化可以在稳定喷雾过程中或在 引用用于高吞吐量实施的校准剂。建立在以前收集的数据基础上的神经网络 将允许持续和轻量级的模型改进。将解决提到的第二个障碍 通过在一维和三维空间中定量识别和放置不稳定的修改的软件。双重的-- 软件平台将对淀粉样β蛋白及其寡聚体进行严格的筛选和定量，从而导致 关于阿尔茨海默病时空进展的数据驱动的结论。 培训将首先通过在Thermo Fisher Science的实习进行。在操作仪器时 硬件和软件要增强高距离信号的传输，申请者将变得熟练 仪器代码和使用方法。然后，申请者将学习如何在 蛋白质组学卓越中心，同时通过Protin精通生物信息学软件。每个人 该中心聚集了从事工业的高影响力科学家，这将促进可概括性设计。 拟议的工作在短期和长期都与NIA的任务直接一致 这意味着什么。一个平台将实现对淀粉样蛋白的高通量、严谨和前所未有的表征 贝塔及其低聚物在其固有的结构背景下。此外，创建的平台将很容易 适用于任何其他疾病或生物系统，无论是否与原始研究无关。结论 断言使用建议的平台应该有助于整个过程中对阿尔茨海默病的理解 进展和大脑区域，希望导致更知情的治疗和治愈。

ABSTRACT The current medical landscape is not equipped to handle Alzheimer’s Disease, with the projected afflicted population tripling between 2010 and 2050 and care costing the U.S. hundreds of billions of dollars each year. To better understand and diagnose the disease, attention has shifted towards biomarkers. The oligomeric forms of amyloid beta—which can be stabilized by metal ions—have been labeled one of the most destructive driving forces behind Alzheimer’s Disease. Mass spectrometry is a prominent force in the disease characterization, but due to the prominent noncovalent landscape of amyloid beta proteoforms, mass spectrometry must be applied in a way that preserves endogenous molecular context for maximal effect. Native top-down mass spectrometry improves on the information accessible from standard approaches by preserving inter- and intramolecular, noncovalent interactions. However, the approach is inaccessible in high throughput due to the required manual nature of data acquisition for large analytes. Another barrier to the characterization of noncovalent protein assemblies via mass spectrometry is the lack of software for the localization of labile modifications (e.g., metals). The proposed work will address the first obstacle through software that autonomously reconfigures the mass spectrometer to enhance each analyte’s transmission for characterization. The optimizations can happen in real time with the actual analyte during steady spray or in reference to a calibrant for high-throughput implementation. Neural networks built on previously collected data will allow for continual and lightweight model improvement. The second obstacle mentioned will be addressed through software that quantitatively identifies and places labile modifications in 1D and 3D space. The dual- software platform will enable rigorous screening and quantitation on amyloid beta and its oligomers, leading to data-driven conclusions on the spatiotemporal progression of Alzheimer’s Disease. Training will first take place via an internship at Thermo Fisher Scientific. While manipulating instrument hardware and software to enhance high-range signal transmission, the applicant will become proficient in instrument code and use. Then, the applicant will learn how to conduct top-down proteomics workflows at the Proteomics Center of Excellence while becoming proficient in bioinformatics software via Protinaceous. Each center is populated with high-impact scientists that engage in industry, which will prompt generalizable design. The proposed works align directly with the NIA’s mission in both their short-term and long-term implications. A platform will enable high-throughput, rigorous, and unprecedented characterization of amyloid beta and its oligomers within their native structural contexts. Furthermore, the created platform will be readily applicable to any other disease or biological system, tangential to the original research or not. The conclusions asserted using the proposed platform should inform understanding of Alzheimer’s Disease throughout progression and brain region, hopefully leading to better-informed treatments and a cure.