"Lung MAP" Atlas Research Center

描述（由申请人提供）：“The LungMap”RFA要求研究中心在一个联盟中工作，建立一个开放获取的分子资源，用于研究人类和小鼠肺的后期发展阶段。我们的研究计划将集中于生成和分析人类和小鼠肺从小管-囊泡晚期到肺泡发育时期的详细的NexGen基因表达数据。肺由不同的实质细胞和造血细胞组成，它们的丰度、分化和功能在发育、区域和物种之间存在差异。不同细胞类型之间的时空相互作用协调了肺的形成和功能。虽然调节肺形成的许多基因和网络在脊椎动物物种之间是共享的，但小鼠和人类肺的生理、结构和区域特异性细胞类型以及基因表达模式存在差异。为了有效地将小鼠模型的数据转化为人类，了解它们在细胞水平上的分子相似性和差异性至关重要。我们将为人类和小鼠肺实质生成详细的细胞特异性RNA数据库。单细胞RNA-seq将使用来自激光捕获显微解剖（LCM）、FACS、RNA分析和分子成像的数据进行解释，以创建人和小鼠肺的表达图谱。计算和“系统生物学”方法将整合这些数据，创建易于访问的数据库，从而协同与肺发育、疾病和功能相关的研究。该应用程序将重点分析正常肺实质细胞(例如，导气管和肺泡上皮细胞；血管内皮细胞，包括静脉、动脉、淋巴和毛细血管细胞

DESCRIPTION (provided by applicant): "The LungMap" RFA tasks Research Centers to work in a consortium to build an open access molecular resource for the study of human and mouse lung during later stages of development. Our research plan will focus to the generation and analysis of detailed NexGen gene expression data for both human and mouse lung from the late canalicular-saccular through the alveolar periods of development. The lung is comprised of diverse parenchymal- and hematopoietic derived cells, whose abundance, differentiation, and functions vary developmentally, regionally, and among species. Temporal and spatial interactions among diverse cell types orchestrate formation and function of the lung. While many of the genes and networks regulating lung formation are shared among vertebrate species, the physiology, structure and regional specific cell types, and gene expression patterns vary between murine and human lung. To effectively translate data from mouse models to humans, it is critical to understand their molecular similarities and differences at the cellular level. We will generate a detailed cell specific RNA database for human and mouse lung parenchyma. Single cell RNA-seq will be interpreted with data derived from laser capture microdissection (LCM), FACS, RNA analyses, and molecular imaging to create an expression map of human and mouse lung. Computational and "systems biology" approaches will integrate these data to create readily accessible databases that will synergize research related to pulmonary development, disease, and function. This application will focus to the analysis of cells from normal lung parenchyma (e.g., conducing airway and alveolar epithelial cells; vascular endothelial cells, including venous, arterial, lymphatic, and capillary cells; stromal and adventitial cells, including fibroblasts, pericytes, myofibroblasts, and cartilage; and neuroepithelial and neuronal cells). Aim 1: Cell Specific NexGen RNA Analyses to Create an "Atlas" of Gene Expression In the Murine Lung. Single cell isolation, FACS purification, and LCM of lung parenchymal cells, and RNA-Seq will create a temporal-spatial map of gene expression in the mouse from E15.5 to postnatal day 28. Aim 2: Cell Specific NexGen RNA Analysis to Create an Atlas of Gene Expression In Human Fetal Lung. Single cell isolation, LCM, and FACS analysis followed by RNA-Seq will create a map of the temporal-spatial patterns of the gene expression during human lung formation from 22 weeks of gestation through the postnatal-alveolar period. Aim 3: Bioinformatics Analysis of NexGen RNAs to Define Gene Expression During Murine and Human Lung Formation. Bioinformatics and computational biology of RNA sequence data from the murine and human lung studies will be analyzed, curated using defined anatomic ontologies, and integrated to provide a sensitive and quantitative measure of gene expression patterns of the precise RNA splice forms, microRNAs and other noncoding RNAs expressed during lung formation. (End of Abstract)