Emergent properties of signaling network degradation that mediate homeostatic failure during aging

信号网络退化的新兴特性介导衰老过程中的稳态失败

• 批准号: 10207412
• 负责人: MATT KAEBERLEIN
• 金额: $ 54.89万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10207412
• 关键词: Age; Aging; Biological Assay; Catalogs; Cell physiology; Cells; Cellular Stress; Computer Models; Data Set; Dependence; Disease; Engineering; Environment; Event; Exposure to; Failure; Gene Deletion; Goals; Health; Homeostasis; Human; Individual; Knowledge; Libraries; Longevity; Maps; Mass Spectrum Analysis; Measurement; Measures; Mediating; Microfluidics; Modeling; Molecular; Organism; Penetrance; Performance; Phosphorylation; Phosphorylation Site; Phosphotransferases; Population; Prevalence; Process; Property; Protein Kinase; Proteins; Proteomics; Reporter; Research; Resolution; Saccharomycetales; Series; Signal Pathway; Signal Transduction; Stress; Structure; System; Systems Biology; Time; Vertebral column; Work; Yeasts; acute stress; age related; aged; base; biological systems; design; environmental change; experience; experimental study; frailty; functional decline; genetic manipulation; healthspan; improved functioning; insight; microfluidic technology; middle age; mortality risk; mutant; network models; phosphoproteomics; predictive test; single cell proteins

PROJECT SUMMARY Loss of homeostatic capacity is a fundamental and defining property of aged organisms from yeast to humans. The global protein kinase-substrate network forms an essential backbone of the homeostatic signaling network (HSN) that allows cells to respond appropriately to a dynamic environment and cellular needs. We propose that aging can be modeled as a series of changes to the HSN that directly impinge on core cellular functions. The overarching goal of this proposal is to model the HSN in yeast, and to understand the mechanisms by which degradation of the HSN results in functional declines and increasing risk of mortality with age. To accomplish this goal we will use a combination of global and targeted mass spectrometry approaches to map at high resolution the kinases and substrates that comprise the HSN. We will then expose aging yeast to perturbations, and measure changes in signaling using phosphoproteomics. Finally, we will combine these population level analyses with single-cell microfluidics measurements in order to define the penetrance and temporal dynamics of key components of the network (kinases and substrates) and reporters of cellular functions that are most prone to degradation throughout aging. This approach will allow us to determine, for the first time, the extent to which individual cells experience distinct aging trajectories. We will use this information to develop models for network degradation with age and to predict key components of the network prone to failure, which could potentially be strengthened to build a more robust network. We will test these predicted improvements by engineering them within yeast strains and assessing whether the strains indeed maintain important network structures with age, keep cellular functions of their youthful state, and, perhaps, live longer.

项目概要 体内平衡能力的丧失是从酵母到人类的衰老生物体的一个基本和决定性的特性。 全局蛋白激酶底物网络构成稳态信号网络的重要支柱 （HSN）使细胞能够对动态环境和细胞需求做出适当的反应。我们建议 衰老可以被建模为 HSN 的一系列变化，这些变化直接影响核心细胞功能。 该提案的总体目标是对酵母中的 HSN 进行建模，并通过以下方式了解其机制： 随着年龄的增长，HSN 的退化会导致功能下降并增加死亡风险。到 为了实现这一目标，我们将结合使用全局和目标质谱方法来绘制地图 以高分辨率显示构成 HSN 的激酶和底物。然后我们将老化酵母暴露于 扰动，并使用磷酸蛋白质组学测量信号传导的变化。最后，我们将把这些结合起来 使用单细胞微流体测量进行群体水平分析，以确定外显率和 网络关键成分（激酶和底物）和细胞报告基因的时间动态 在衰老过程中最容易退化的功能。这种方法将使我们能够确定，对于 第一次，单个细胞经历不同的衰老轨迹的程度。我们将使用这个 用于开发网络随时间退化的模型并预测网络关键组件的信息 容易出现故障，这可能会得到加强，以建立一个更强大的网络。我们将测试这些 通过在酵母菌株中进行改造并评估这些菌株是否确实可以预测改进 随着年龄的增长，维持重要的网络结构，保持年轻状态的细胞功能，也许， 活得更久。