Monitoring the Aging Lung Through Proteomics and Quantum Imaging

肺组织的长期健康与蛋白质折叠的可持续性及其对肺组织的影响密不可分。 与细胞的能量健康密切相关的功能。这是通过新兴的范式实现的， 蛋白质稳态或蛋白质稳态，一组整合的ATP依赖性生物途径， 产生并维持蛋白质组。蛋白质稳态平衡蛋白质的生物合成，折叠，易位， 蛋白质复合物组装和/或拆卸和清除的挑战， 外源性折叠应力是对局部物理环境和老化的反应。我们建议， 蛋白质稳态网络（PN）对正常衰老和随年龄增长的环境损伤的反应 挑战老化的肺的折叠健康。在核心蛋白B中，我们建议定量测量 肺上皮细胞，以了解“顺应性”，反映了年轻肺的抵抗力的生理学 环境挑战，“储备”，肺的诱导缓冲能力， 日常/慢性挑战，以及“脆弱”-导致对损失的敏感性增强的生理变化 所有这些都增加了个人和老年人的临床/经济负担。 医疗保健系统。在具体目标1中，核心B将建立对 蛋白质组和蛋白质稳态环境对正常衰老和年轻人对甲型流感感染的反应 老老鼠这将提供一个基线，以解决正常老化和病毒干扰的折叠/老化健康 项目1-3中列出的问题。核心B将了解不断变化的蛋白质稳定健康 在老化过程中，通过系统应用质谱（MS）来定量表征 在小鼠的整个正常和受挑战的寿命期间， 无标记多维蛋白质鉴定技术（MudPIT）的应用，串联质量标记 (TMT)MS和信号离子反应（SRM）技术。目标1中的研究将与使用 Aim 2中新开发的生物传感器成像技术，可测量蛋白质折叠的整体状态 健康的肺泡上皮细胞和肌肉在系统和定量的方式。生物传感器使我们能够 使用量子记录实时跟踪小鼠整个寿命期间的折叠健康状态 CIVIS-K卡尺的性能。成像技术是基于新的和新兴的蛋白质稳态 这些原则预计将强烈影响我们对肺泡正常健康范围的理解， 上皮环境和肌肉以及它们对压力挑战如甲型流感的反应。而Aim 1严格量化的颗粒特征的蛋白质稳态程序在老化过程中，目标2图像生活如何 蛋白质稳态计划作为一个综合单位发挥作用，以保护青年人的肺，我们建议的活动是 老化过程中受损。核心B将通过执行以下工作，为项目1-3中的每一项工作做出贡献： 整合了旨在解决衰老过程中蛋白质稳定性脆弱性的机械项目目标的分析。

The long-term health of pulmonary tissue is inextricably linked to the sustainability of the protein fold and its function that is tightly coupled to the energetic health of the cell. This is achieved by the emerging paradigm of protein homeostasis or proteostasis, a collection of integrated ATP-dependent biological pathways that generate and maintain the proteome. Proteostasis balances protein biosynthesis, folding, translocation, protein complex assembly and/or disassembly and clearance with the challenges imposed by endogenous and exogenous folding stress in response to the local physical environment and aging. We suggest that changes in the proteostasis network (PN) in response to normal aging and environmental insults that accrue with age challenge the folding health of the aging lung. In Core B, we propose to quantitatively measure proteostasis in the lung epithelium to understand `resilience', the physiology that reflects the resistance of younger lungs to environmental challenges, `reserve', the inducible buffering capacity of the lung that protects against daily/chronic challenges, and `frailty'- the physiologic changes that underlie the enhanced susceptibility to loss of healthspan, all of which contribute to the clinical/financial burden of aging for the individual and the healthcare system. In Specific Aim 1, Core B will establish a rigorous understanding of the response of the proteome and proteostasis environment to normal aging and in response to the influenza A infection in young and old mice. This will provide a baseline to address the normal aging and viral perturbed folding/aging health questions outlined in Projects 1-3. Core B will achieve an understanding of the changing proteostatic health program during aging by systematic application of mass spectrometry (MS) to quantitatively characterized the proteostasis environment of the lung throughout the normal and challenged lifespan of the mouse through application of label-free Multidimensional Protein Identification Technology (MudPIT), tandem mass tagging (TMT) MS and Signal Ion Reaction (SRM) technologies. The studies in Aim 1 will be integrated with the use of newly developed biosensor imaging technologies in Aim 2 that measure the global state of protein folding health in the alveolar epithelium and muscle in a systematic and quantitative fashion. Biosensors allow us to follow in real-time the state of folding health throughout a mouse lifespan using the quantum recording capabilities of the Caliper CIVIS-K. The imaging technology is based on new and emerging proteostasis principles that are expected to strongly impact our understanding of the normal healthspan of the alveolar epithelial environment and muscle and their response to stress challenges such as influenza A. Whereas Aim 1 rigorously quantifies the granular features of the proteostasis program during aging, Aim 2 images live how the proteostasis program functions as an integrated unit to protect the lung in youth, activities we proposed are compromised during aging. Core B will contribute to the efforts of each of the Projects 1-3 by performing analyses that integrate mechanistic Project Aims designed to address proteostatic frailty during aging.