Multiplex Mass Spectrometric Protein Assays for Precise Monitoring of the Pathophysiology of Obesity

用于精确监测肥胖病理生理学的多重质谱蛋白质分析

• 批准号: 10020391
• 负责人: Wei-Jun Qian
• 金额: $ 75.56万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10020391
• 关键词: Adopted; Adult; Affinity; Anti-Inflammatory Agents; Area; Benchmarking; Biochemistry; Biological Assay; Biological Markers; Blinded; Blood Proteins; Body Weight decreased; Buffaloes; C-reactive protein; Cardiovascular Diseases; Child; Clinical; Collaborations; Communities; Coupled; Data; Detection; Diabetes Mellitus; Disease; Epidemic; Failure; Family; Functional disorder; GDF15 gene; Goals; Hormones; IGF1 gene; Immunoassay; Individual; Inflammatory; Insulin; Insulin Resistance; Insulin-Like Growth-Factor-Binding Proteins; Intelligence; LCN2 gene; Laboratories; Leptin; Mass Spectrum Analysis; Matrix Metalloproteinases; Medicine; Metabolism; Monitor; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Overweight; Patient Care; Peptides; Performance; Phase; Plasma; Prevalence; Preventive Intervention; Procedures; Protein Isoforms; Proteins; Protocols documentation; RBP4 gene; Reagent; Reporting; Reproducibility; Research; Research Institute; Resolution; Sampling; Site; Solid; Specificity; Standardization; Technology; Translational Research; Translations; Universities; Validation; Vermont; Weight; Work; adipokines; adiponectin; assay development; base; biomarker panel; blood glucose regulation; cancer type; clinical application; cohort; combat; data warehouse; disorder prevention; energy balance; experience; experimental study; ghrelin; inflammatory marker; insight; mortality risk; multiplex detection; personalized care; personalized medicine; precision medicine; pressure; prohormone; resistance exercise; validation studies

PROJECT SUMMARY/ABSTRACT The prevalence of overweight and obesity has increased dramatically in recent decades in both children and adults, reaching an epidemic proportion. In order to gain new insights into the pathophysiology of obesity towards better personalized patient care, there is a critical need to develop reliable multiplex protein assays that can be easily implemented in clinical laboratories to enable precise monitoring of many hormones and inflammatory markers closely associated with obesity. Unfortunately, current clinical assays often lack of reproducibility, especially between different clinical laboratories due to the lack of assay standardization. Targeted mass spectrometry is a promising technology for providing robust multiplex assays for blood proteins. Therefore, the overall objective of this application is to develop and validate reproducible and transferable mass spectrometry-based multiplex protein assays for enabling precise quantification of a panel of obesity markers. The panel will cover multiple aspects of obesity pathophysiology, including glucose homeostasis and energy balance (e.g., insulin, leptin, and other hormones), pro-inflammatory markers (e.g., C-reactive protein), and anti-inflammatory markers (e.g., adiponectin). To facilitate full validation of the robustness and transferability of the assays, an inter-lab assay validation will be pursued. Specifically, Aim 1 will be focused on initial assay development for optimal configurations and on demonstrating confident multiplex detection of endogenous analytes in blood plasma. Aim 2 will be centered on assay optimization and full assay characterization in the aspects of reproducibility, stability, selectivity, linearity, and limit of quantification. Aim 3 will demonstrate the utility of the assays through inter-lab quantification of endogenous analytes in a pilot cohort of clinical plasma samples from normal weight and obese subjects, and obese individual before and after weight loss and benchmarking against well-established immunoassays for selected analytes such as insulin and leptin. To facilitate the easy- implementation of the validated assays in other laboratories, all assay data including chromatographic data and detailed standard operating procedures will be shared through public data repositories and assay portals. Together, the project will establish robust and easy-to-transfer multiplex assays that will enable precise monitoring of the pathophysiology of obesity.

项目摘要/摘要 最近几十年，在儿童和 成人，达到流行比例。为了获得对肥胖病理生理学的新见解 为了更好地个性化的患者护理，迫切需要开发可靠的多重蛋白质测定法 可以在临床实验室中轻松实施，以便对许多激素和 炎症标记与肥胖密切相关。不幸的是，当前的临床测定通常缺乏 可重复性，特别是由于缺乏测定标准化而在不同的临床实验室之间。 靶向质谱是一种有前途的技术，用于为血液蛋白提供鲁棒的多重测定。 因此，本应用的总体目标是开发和验证可再现且可转移的质量 基于光谱的多重蛋白质测定法，以实现肥胖标记面板的精确定量。 该面板将涵盖肥胖病理生理学的多个方面，包括葡萄糖稳态和能量 平衡（例如胰岛素，瘦素和其他激素），促炎性标记（例如C反应蛋白）和 抗炎标记（例如脂联素）。促进充分验证鲁棒性和可转移性 在测定中，将进行LAB间测定验证。具体而言，AIM 1将重点放在初始测定上 开发最佳配置并展示内源性的自信多重检测 血浆分析物。 AIM 2将以测定优化和完整的测定为中心 可重复性，稳定性，选择性，线性性和定量极限的各个方面。 AIM 3将证明 通过LAB间定量内源性分析物在临床等离子体中的内源性分析物的实用性 体重正常和肥胖受试者的样本，以及体重减轻之前和之后的肥胖个人 针对胰岛素和瘦素等选定分析物的公认的免疫测定基准测试。到 促进在其他实验室中轻松实施经过验证的测定法，所有测定数据包括 色谱数据和详细的标准操作程序将通过公共数据共享 存储库和测定门户。该项目将共同建立强大且易于转移的多重测定 这将实现对肥胖病理生理学的精确监测。