Immunobiology and alveolar physiology of the aging lung

项目摘要 预计老年人口在未来40年将翻一番，因此与老龄化有关的疾病 很可能成为主要的医疗负担。慢性阻塞性肺疾病，包括肺气肿 肺部疾病和间质性肺疾病，以及包括急性肺损伤(ALI)在内的一般肺部病理 和相关的急性呼吸窘迫综合征(ARDS)，在老年人中急剧增加，尽管 根本原因和机制仍未确定。重要的是，衰老与许多 肺生理和肺免疫的变化，包括肺活量减少，粘液纤毛减少 清除，并增加对细菌和病毒感染的易感性。肺生理和肺功能的改变 病原体的易感性可能是相关的；然而，关于这些与年龄相关的影响，我们知之甚少。 免疫系统监测肺的生理变化。识别这些基因的分子变化 随着年龄的增长，可以导致新的治疗干预措施，以帮助保护老化的肺和减少 疾病发病率。我们已经建立了一种新的人类组织资源，在这种资源中我们获得了多个淋巴 来自所有年龄段的器官捐赠者的粘膜组织，包括所有成年至第九个十年的年龄段。 重要的是，我们获得了肺、肺相关和外周淋巴组织，使衰老的新研究成为可能。 在几十年的连续统一体中发生的相关变化。我们的中心假设是，随着年龄的增长， 肺内常驻免疫细胞和肺相关淋巴的免疫监视和保护， 导致肺细胞完整性降低，对损伤的易感性增加。在建议的研究中，我们会 采用多学科方法研究肺免疫系统与肺之间的相互作用 上皮细胞是年龄的一种功能。在目标1中，我们将确定肺常驻免疫细胞和肺相关 淋巴结(LN)随年龄变化。我们将使用高维细胞和分子轮廓，使用High 参数流式细胞术和整个转录组图谱以评估免疫细胞群体的变化，以及 免疫荧光成像以确定肺部常驻免疫细胞的频率和位置，以及 通过对LN卵泡的检查，了解LN的功能活性。在目标2中，我们将使用实时光学成像 以确定年龄对肺泡-毛细血管液体屏障质量的影响， 肺泡上皮中线粒体的功能，以及常驻AM维持缝隙连接的程度 与肺泡上皮细胞的旁分泌交通。在目标3中，我们将定义年龄驱动基因 在所有肺细胞中表达程序，使用单细胞RNA-Seq来评估基因表达程序是如何 定义肺上皮细胞、内皮细胞、先天免疫细胞和获得性免疫细胞会改变和动力学 这些变化。拟议的研究将揭示对人类肺部免疫学和生理学的新见解。 以及与年龄相关的变化的机制，这些变化使个人容易患上更多的呼吸道疾病。

Project Summary The elderly population is expected to double in the next 40 years, and therefore aging-related diseases are likely to become a major healthcare burden. Lung diseases, including emphysema, chronic obstructive pulmonary disease, and interstitial lung disease, and general lung pathologies including acute lung injury (ALI) and the associated acute respiratory distress syndrome (ARDS), increase dramatically in the elderly, although the underlying causes and mechanisms remain undefined. Importantly, aging is associated with a number of changes in lung physiology and lung immunity, including a decreased vital capacity, decreased mucociliary clearance, and increased susceptibility to both bacterial and viral infections. Changes in lung physiology and pathogen susceptibility may be related; however, there is little known regarding the impact of these age-related physiologic changes on surveillance of the lung by the immune system. Identifying molecular changes in these processes with age can lead to new therapeutic interventions to help protect the aging lung and reduce disease incidence. We have established a novel human tissue resource where we obtain multiple lymphoid and mucosal tissues from organ donor of all ages, including all ages of adulthood up to the 9th decade of life. Importantly, we obtain lungs, lung-associated and peripheral lymphoid tissues, enabling novel study of aging- associated changes over a continuum of decades. Our central hypothesis is that with age, there is decreased immune surveillance and protection both from lung-resident immune cells and lung-associated lymph nodes, resulting in decreased lung cell integrity and increased susceptibility to damage. In the proposed study, we will take a multi-disciplinary approach to study the interaction between the lung immune system and lung epithelium as a function of age. In aim 1 we will identify how lung resident immune cells and lung-associated lymph nodes (LN) alter with age. We will use high dimensional cellular and molecular profiling using high parameter flow cytometry and whole transcriptome profiling to assess changes in immune cell populations, and immunofluorescence imaging to determine the frequency and localization of lung-resident immune cells, and the functional activity of LN through examination of LN follicles. In aim 2, we will use real time optical imaging of the human lung to determine the effect of age on the quality of the alveolar-capillary fluid barrier, mitochondrial function in the alveolar epithelium, and the extent to which resident AMs maintain gap junctional and paracrine communication with the alveolar epithelium. In aim 3, we will define the age-drive gene expression program in all lung cells, using single cell RNA-Seq to assess how gene expression programs that define lung epithelial cells, endothelial cells, innate and adaptive immune cells will be altered and the kinetics of these alterations. The proposed studies will reveal new insights into human lung immunology and physiology and mechanisms for age-associated changes that predispose individuals to increased respiratory disease.