Mapping Age-Related Changes in the Lung

摘要 衰老与肺部疾病的患病率增加有关，例如特发性肺纤维化（IPF）、 然而，慢性阻塞性肺疾病（COPD）和急性肺损伤（ALI） 导致与年龄相关的肺部疾病发病机制的衰老过程的机制尚未确定 已被完全阐明。 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。基于上述，我们建议创建一个人类衰老肺图谱（AHAL），其中包括 各个肺细胞类型的转录组图。这将填补现有的知识空白并提供丰富的知识 未来研究的资源。转录差异可能反映基因组不稳定性、压力变化 线粒体功能障碍、衰老和 SASP 标记物的反应和富集。我们也 研究线粒体自噬缺陷与衰老肺部衰老相关的假设。 根据我们和其他研究，我们假设原发性自分泌年龄相关的衰老有所不同 从与年龄相关的肺部疾病中观察到的次级旁分泌衰老程序转录而来 例如IPF 我们提出 3 个独立的目标来定义肺部与年龄相关的变化 目标 1：绘制人类肺部转录组与年龄相关的变化并对其进行表征。我们将表演 来自健康年轻人和老年人的单细胞 RNA 序列 (scRNAseq) 和批量 RNA 序列 (RNAseq) 人类肺部并建立了每种肺细胞类型以及细胞的转录组变化的综合图谱 肺部老化的人口变化。为了了解实际年龄和功能年龄有何不同，我们将 还将已确定的与年龄相关的基因特征与 IPF 上叶和下叶的基因特征进行比较，IPF 是一种年龄相关的基因特征。 相关肺部疾病。数据、结果以及肺细胞和转录组图谱将公开 通过我们将开发的基于云的综合门户网站。 目标 2. 表征线粒体自噬在衰老肺和衰老表型中的作用。我们会 研究增强线粒体自噬可减轻衰老肺中线粒体功能障碍的假设 并调节衰老表型。我们已经鉴定出线粒体去泛素酶 USP30， 线粒体自噬的负调节因子，是线粒体自噬下降的关键因素。作为概念证明，我们将 使用原代小鼠研究 USP30 在细胞衰老发展中的潜在致病作用 以及人类细胞和 USP30-/- 小鼠。 目标 3. 识别衰老肺细胞区室中与衰老相关的变化。我们将研究如何 老化的衰老细胞及其 SASP 库在细胞区室之间存在差异，并且对组织有贡献 受伤后的重塑。我们将对年轻和老年肺部的分离衰老细胞进行 scRNAseq 表征其细胞特异性衰老转录组。我们还将调查与年龄相关的衰老是否是 与 IPF 肺部细胞中观察到的衰老相似。！

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.!