Normal Aging Lung Cell Atlas (NALCA)

项目总结 《正常衰老肺细胞图谱》(NALCA)的总体目标，我们对RFA-HL-19-012的反应 (解密人类肺老化的分子景观1)，是为了生成一个概要 在肺老化过程中发生的mRNA、microRNA和表观遗传标记的动态细胞特异性变化 关注单个细胞和肺泡微环境。我们计划使用这个概要来生成一个动态的 正常人肺老化的时间调控模型。这个应用程序的调查人员有重要的 在肺基因组学、表观基因组学、生物工程、衰老、肺泡发育、系统和 计算生物学。这一提议的前提是资源的可用性，包括正常的人力资源 不同年龄、衰老的野生型以及与衰老相关的转基因小鼠的肺，在 高通量技术包括单细胞RNAseq、激光捕获显微切割、甲基化分析、 蛋白质组学和组织工程，以及在开发分析方法方面的成熟记录 剖析时间监管网络。它也在很大程度上受益于与其他非重叠的协同效应 NIH-NHLBI和NIH-NIA由共同调查人员领导的这项提案。我们将解决这些目标 本项目的具体目标如下：具体目标1.确定特定的细胞和微环境 人类肺老化过程中编码和非编码RNA的变化。我们将执行所有单细胞RNASEQ 人类肺中的细胞跨越了正常衰老的光谱，利用一组独特的老化肺。 LCM RNAseq对肺微环境的分析将在相同的肺上并行执行，分析 肺泡、毛细血管和炎性浸润物的改变。老化甲基化标记与细胞特异性验证 也将上演。具体目标2.使用详细的时间分析确定肺老化的关键时间点 对小鼠肺老化的影响。我们将对不同性别的野生型小鼠的肺进行单细胞RNASEQ 确定基因表达发生关键变化的确切时间点的时间点。然后我们将分析 这些时间点的细节，利用了寿命和衰老机制改变的转基因小鼠。 蛋白质组学分析将在重叠的老鼠和人类关键时间点上进行。具体目标3.至 建立一个全面的肺老化转录动态调控多细胞模型。我们将使用和 扩展我们的分析工具以对特定的动态信号、监管网络和蜂窝交互进行建模，请访问 单细胞水平在肺老化中的作用。该模型将提供见解，并将与科学界共享， 通过一个称为老化肺动态调节多细胞模型的高度交互和直观的网络界面 (AgeDREMM)，这将允许可视化和共享我们的结果。

PROJECT SUMMARY The overall objective of the ‘Normal Aging Lung Cell Atlas’ (NALCA), our response to RFA-HL-19-012 (Deciphering the Molecular Landscape of Lung Aging in Humans UO1), is to generate a compendium of the dynamic cell specific changes in mRNA, microRNA, and epigenetic marks that happen during lung aging with a focus on single cells and the alveolar microenvironment. We plan to use this compendium to generate a dynamic temporal regulatory model of normal human lung aging. The investigators on this application have significant expertise in lung genomics, epigenomics, bioengineering, aging, alveolar development, systems and computational biology. The premise of this proposal lies in the availability of resources, including normal human lungs at different ages, aged wild-type as well as aging relevant genetically modified mice, unique expertise in high throughput technologies including single cell RNAseq, laser capture microdissection, methylation profiling, proteomics and tissue engineering, and proven track record in developmental of analytical approaches that dissect temporal regulatory networks. It is also largely benefitting from synergisms with other non-overlapping NIH-NHLBI and NIH-NIA projects headed by the coinvestigators on this proposal. We will address the objectives of this project by the following specific aims: Specific Aim 1. To identify cell and microenvironment specific changes in coding and non-coding RNAs across human lung aging. We will perform single cell RNAseq of all the cells in the human lung across the spectrum of normal aging utilizing a unique collection of aging lungs. Analysis of lung microenvironments by LCM RNAseq will be performed in parallel on the same lungs, analyzing changes in alveoli, capillaries and inflammatory infiltrates. Aging methylation markers and cell specific validations will also be performed. Specific Aim 2. To identify key time points in lung aging using detailed temporal analysis of mouse lung aging. We will perform single cell RNAseq of lungs of wild-type mice of both sexes at multiple time points to identify the exact time points in which key changes in gene expression occur. We will then analyze these time points in detail, utilizing genetically modified mice with altered lifespan and aging mechanisms. Proteomic analysis will be performed on overlapping mouse and human key time points. Specific Aim 3. To develop a comprehensive transcriptional dynamic regulatory multicellular model of lung aging. We will use and extend our analytical tools to model specific dynamic signaling, regulatory networks, and cellular interactions, at the single cell level in lung aging. The model will provide insights and will be shared with the scientific community, through a highly interactive and intuitive web interface called Aging Lung Dynamic Regulatory Multicellular Model (AgeDREMM) that will allow visualization and sharing of our results.