Characterization of the Various Senescent Cells in the Aging Lungs

衰老肺中各种衰老细胞的表征

基本信息

批准号：
9086628
负责人：
Claude Le Saux
金额：
$ 19.81万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9086628
关键词：
Address Affect Age Aging Aging-Related Process Alveolar Alveolar Cell Animal Model Area Automobile Driving Brain CDKN2A gene Cell Aging Cell Separation Cells Characteristics Chronic Obstructive Airway Disease Collaborations Data Detection Diagnosis Disease Documentation Dose Elderly Endothelial Cells Epithelial Epithelial Cells Extracellular Matrix Fibroblasts Flow Cytometry Fluorescence Future Ganciclovir Growth Hamman-Rich syndrome Health Human In Vitro Incidence Individual Kidney Laboratories Letters Life Expectancy Link Lung Lung diseases Modeling Modification Molecular Mus Onset of illness Organ Pathway interactions Phenotype Physiological Play Population Positioning Attribute Process Proteins Pulmonary Fibrosis Reporter Role Sampling Signal Pathway Societies Structure of parenchyma of lung System TP53 gene Testing Therapeutic Tissues Transcript Transgenic Mice Transgenic Organisms Work Wound Healing age related aged base body system cell type differential expression disability empowered health care service in vivo injured injury and repair insight irradiation mortality mouse model new therapeutic target nonhuman primate protein expression public health relevance response to injury senescence

项目摘要

DESCRIPTION (provided by applicant): Despite ample documentation of age-related changes in physiologic parameters, mechanisms governing the process of aging of the lungs have not been well studied. Among the identified age-associated changes is the modification of the extracellular matrix (remodeling). These changes result in alteration of the aged lungs response to injury and repair. As importantly, the number of senescent cells in the lungs increases with age. However, the mechanisms by which these cells provoke aging of the lungs are uncertain. All senescent cells are in growth arrest but continue to secrete large amounts of proteins that affect their microenvironment. Seemingly contradictory data indicate that, on the one hand, senescent fibroblasts are beneficial in wound healing but, on the other hand, limiting the number of senescent alveolar epithelial cells is beneficial in pulmonary fibrosis. Thus, there is a critical need to characterize the senescence profile of the various cells present in the lungs (mainly endothelial, epithelial, and fibroblast cells) to determine their relative contribution to he aging process. Despite the detection of senescent cells in aged lungs, their role in physiological remodeling associated with aging is unknown and their secretory phenotype remains undefined. Studying senescence in vivo has been technically challenging; however, we have recently obtained the p16-3MR transgenic system that empowers in vivo identification of p16ink4A+ cells (senescent cells) and their subsequent isolation by flow cytometry as well as their conditional deletion via the administration of gancyclovir. We propose to use this mouse model to more definitely define the role of senescent cells in lung aging; we will validate our data in nonhuman primate aging lungs, a translational model to human aging. We hypothesize that each senescent cell type (alveolar epithelial, endothelial and fibroblasts) has a characteristic secretory associated senescence profile to indicate their relative contribution to aging. Two specific aims are proposed to address this hypothesis: 1) To identify the senescence-associated secretory profile of each different senescent cell type (alveolar epithelial and endothelial cells and fibroblasts) to determine their contribution to lung tissue remodeling. Senescent and non-senescent cells will be isolated from non-lethally irradiated p16-3MR mouse lungs by flow cytometry and their transcriptional profiling will be determined. The expression of differentially expressed identified proteins will be validated in young and aged nonhuman primate lungs; and 2) To determine the contribution of senescent cells in the age-associated remodeling of the lungs. Using the p16-3MR transgenic mouse, we will determine whether the presence of senescent cells affects the remodeling process in the lungs by deleting p16ink4A+ cells after non-lethal dose irradiation. The data gathered with this project will characterize the secretory profile of various senescent cell types that have an important functional role in remodeling in the lungs but also in other organs such as the brain or the kidney (senescent endothelial cells and fibroblasts) and provide new information as to mechanism.

描述（由适用提供）：尽管有足够的文献记录了与年龄相关的生理参数变化，但管理肺部衰老过程的机制并没有很好地研究。在确定的与年龄相关的变化中，是细胞外基质（重塑）的修饰。这些变化导致老年肺对损伤和修复的反应改变。重要的是，肺中的感觉细胞的数量随着年龄的增长而增加。但是，这些细胞引起肺衰老的机制尚不确定。所有感觉细胞都处于生长停滞状态，但继续向影响其微环境的大量蛋白质进行秘密。看似矛盾的数据表明，一方面，感觉成纤维细胞有益于伤口愈合，但另一方面，限制感觉肺泡上皮细胞的数量对肺纤维化有益。这是迫切需要表征存在肺中各种细胞的感应谱（主要是内皮，上皮和成纤维细胞），以确定它们对衰老过程的相对贡献。尽管检测到老年肺中的敏感细胞，但它们在与衰老相关的物理重塑中的作用尚不清楚，并且其秘书表型仍然不确定。在技术上挑战了体内的感受。但是，我们最近获得了P16-3MR转基因系统，该系统赋予了P16INK4A+细胞（感觉细胞）的体内鉴定以及通过流式细胞仪及其条件缺失通过GangyClovir的给药进行分离。我们建议使用该小鼠模型更清楚地定义感觉细胞在肺部衰老中的作用。我们将在非人类灵长类动物衰老肺中验证我们的数据，这是一个翻译成人类衰老的模型。我们假设每种感觉细胞类型（牙槽上皮，内皮和成纤维细胞）具有特征性的秘密相关感应谱，以表明它们对衰老的相对贡献。提出了两个具体目的来解决这一假设：1）确定每种不同的感应细胞类型（牙槽上皮细胞和内皮细胞和成纤维细胞）的感应相关的秘书概况，以确定它们对肺组织重塑的贡献。通过流式细胞仪，将从非辐照的p16-3MR小鼠肺中分离出衰老和非传染细胞，并确定其转录分析。在年轻和年龄的非人类灵长类肺中将验证不同表达的蛋白质的表达。 2）确定在与年龄相关的肺重塑中的感觉细胞的贡献。使用p16-3MR转基因小鼠，我们将通过删除非致命剂量照射后删除P16INK4A+细胞来确定感觉细胞的存在是否会影响肺部的重塑过程。与该项目一起收集的数据将表征各种感觉细胞类型的秘书概况，这些细胞类型在重塑中具有重要功能作用肺部，以及其他器官，例如大脑或肾脏（感觉内皮细胞和成纤维细胞），并提供有关机制的新信息。