Normal Aging Lung Cell Atlas (NALCA)

项目摘要 “正常老化肺细胞图谱”（NALCA）的总体目标，我们对RFA-HL-19-012的回应 （破译人类肺衰老的分子景观UO 1），是为了产生一个纲要， mRNA、microRNA和表观遗传标记的动态细胞特异性变化发生在肺老化过程中， 关注单个细胞和肺泡微环境。我们计划利用这个纲要来产生一个动态的 正常人肺老化的时间调节模型。这份申请的调查人员对 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 计算生物学这一建议的前提在于资源的可用性，包括正常的人力资源。 肺在不同年龄，老龄野生型以及老龄相关的转基因小鼠，独特的专业知识， 高通量技术，包括单细胞RNAseq、激光捕获显微切割、甲基化谱分析， 蛋白质组学和组织工程，以及在分析方法的发展方面的良好记录， 剖析时间调控网络。它也在很大程度上受益于与其他非重叠的 NIH-NHLBI和NIH-NIA项目由该提案的共同研究者领导。我们将致力于 本项目的具体目标如下：具体目标1。识别细胞和微环境特异性 人类肺老化过程中编码和非编码RNA的变化。我们将对所有人进行单细胞RNAseq 利用老化肺的独特集合，在正常老化的范围内的人肺中的细胞。 通过LCM RNAseq对肺微环境的分析将在相同的肺上平行进行，分析 肺泡、毛细血管和炎性浸润的变化。老化甲基化标志物和细胞特异性验证 也将被执行。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 小鼠肺老化的证据我们将在多个时间点对两种性别的野生型小鼠的肺进行单细胞RNAseq。 时间点，以确定基因表达发生关键变化的确切时间点。我们将分析 这些时间点的细节，利用基因改造的小鼠与改变寿命和老化机制。 将在重叠的小鼠和人关键时间点进行蛋白质组学分析。具体目标3。到 开发一个全面的转录动态调节肺衰老的多细胞模型。我们将使用和 扩展我们的分析工具，以模拟特定的动态信号，调节网络和细胞相互作用， 肺老化的单细胞水平。该模型将提供见解，并将与科学界分享， 通过一个高度互动和直观的网络界面，称为老化肺动态调节多细胞模型， （CMDREMM），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.