Mapping Age-Related Changes in the Lung

摘要 老龄化与肺部疾病的增加有关，如特发性肺纤维化(IPF)， 然而，慢性阻塞性肺疾病(COPD)和急性肺损伤(ALI)的分子水平 衰老过程对年龄相关性肺部疾病发病机制的影响尚未见报道 已经完全阐明了。 在细胞水平上，对定义不同肺细胞健康衰老的因素的了解有限 可用。在此基础上，我们建议建立一个人类衰老肺图谱(Ahal)，该图谱包括 单个肺细胞类型的转录图谱。这将填补现有的知识缺口，并提供丰富的 未来研究的资源。转录差异可能反映了基因组的不稳定、压力的变化 线粒体功能障碍、衰老和SASP标志物的反应和丰富。我们也 研究吞丝分裂缺陷与衰老肺的相互联系的假说。 基于我们的研究和其他研究，我们假设初级自分泌与年龄相关的衰老是不同的。 在年龄相关性肺部疾病中观察到的次级旁分泌衰老程序的转录 例如IPF 我们提出了三个独立的目标来定义与年龄相关的肺部变化。 目的1：定位和描述人类肺中转录的年龄相关变化。我们将表演 健康青年和老年人的单细胞RNA序列(ScRNAseq)和本体RNA序列(RNAseq) 并建立了每种肺细胞类型以及细胞转录变化的全面图谱 老龄化肺部的人口转移。为了了解时间和功能年龄的不同，我们将 还将已识别的与年龄相关的基因签名与IPF的上叶和下叶的签名进行比较，年龄- 相关的肺部疾病。数据、结果以及肺细胞和转录组图谱将公开提供 通过一个基于云的综合门户网站，我们将开发。 目的2.探讨线粒体吞噬在衰老肺和衰老表型中的作用。我们会 探讨线粒体吞噬功能增强减轻衰老肺线粒体功能障碍的假说 并调节衰老表型。我们已经鉴定出线粒体去泛素酶USP30，一个 丝裂原吞噬的负调节因子，是吞丝下降的关键因素。作为概念验证，我们将 利用原代小鼠研究USP30在细胞衰老过程中的潜在致病作用 以及人类细胞和USP30-/-小鼠。 目的3.确定衰老相关的肺细胞隔室的变化。我们将研究如何 衰老的细胞及其SASP谱系因细胞隔间不同而不同，对组织有贡献 损伤后的重塑。我们将在年轻和老年肺的分离衰老细胞中进行scRNAseq和 描述它们细胞特有的衰老转录组。我们还将调查与年龄相关的衰老是否 类似于观察到的IPF肺细胞的衰老。

ABSTRACT Aging is associated with increased prevalence of lung diseases, such as idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), and acute lung injury (ALI), however, the molecular mechanisms of the aging process that contribute to the pathogenesis of age-related lung diseases have not been completely elucidated. At a cellular level, only limited knowledge of the factors that define healthy aging of different lung cells is available. Based on the above, we propose to create an atlas of human aging lung (AHAL) consisting of transcriptomic maps of individual lung cell types. This will fill an existing knowledge gap and provide a rich resource for future research. Transcriptional differences might reflect genomic instability, alterations in stress responses and enrichment of markers of mitochondrial dysfunction, senescence and SASP. We also investigate the hypothesis that defective mitophagy is interconnected with senescence in the aging lung. Based on ours and other studies, we hypothesize that primary autocrine age-related senescence differs transcriptionally from the secondary paracrine senescence program observed in age-related lung diseases such as IPF We are proposing 3 independent aims to define age-related changes in the lung Aim 1: To map and characterize transcriptomic age-related changes in human lungs. We will perform single cell RNA sequence (scRNAseq) and bulk RNA sequence (RNAseq) from healthy young and aged human lungs and build a comprehensive atlas of transcriptomic changes in each lung cell type, as well as cell population shifts in aging lungs. In order to understand how chronological and functional ages differ, we will also compare the identified age-related gene signatures with those of the upper and lower lobe in IPF, an age- related lung disease. Data, results and lung cellular and transcriptome maps will be made publicly available through a cloud based comprehensive web portal that we will develop. Aim 2. To characterize the role of mitophagy in the aging lung and the senescence phenotype. We will investigate the hypotheses that augmenting mitophagy attenuates mitochondrial dysfunction in the aging lung and modulates the senescence phenotype. We have identified the mitochondrial deubiquitinase USP30, a negative regulator of mitophagy, as a key factor for mitophagy decline. As a proof of concept, we will investigate the potential pathogenic role of USP30 in the development of cell senescence using primary murine and human cells and USP30-/- mice. Aim 3. To identify senescence-related changes in aging lung cell compartments. We will examine how aging senescent cells and their SASP repertoire differ between cell compartments and contribute to tissue remodeling after injury. We will perform scRNAseq in isolated senescent cells from young and aged lungs and characterize their cell-specific senescent transcriptome. We will also investigate if age-related senescence is similar to senescence observed in cells from IPF lungs.!