PROJECT ABSTRACT Alveologenesis characterizes the later stages of normal human lung development and is the final step of lung maturation. Alveologenesis is a dynamic, coordinated process that requires the accurate spatial and temporal integration of signals to develop the intricate alveolar structure. While important progress has been made, significant knowledge gaps remain in our understanding of the molecular mechanisms underlying alveologenesis. For this reason, NHLBI established Research Centers (RCs) to create a molecular atlas of the developing lung (LungMAP) focused from birth up to early childhood (~8 years). In the first phase of LungMAP, our RC developed resources that enabled a much more detailed understanding of normal lung development up to early childhood. In LungMAP Phase 2, we will apply our successful approaches and our newest technologies to extend analysis of lung development into early adulthood as well as human disease; focusing on bronchopulmonary dysplasia (BPD), the most common morbidity of preterm infants which is characterized by delayed or deficient lung maturation. Within the lung, the relationship between space, anatomy, and function is fundamental. Therefore, our approach includes new unbiased 3D quantitative imaging approaches implemented with high spatial resolution, as well as cell-specific omics: proteomics (including activity-based proteomics and phosphoproteomics), lipidomics, and metabolomics. The integration of these complementary data collection methods facilitates the establishment of a cell-specific spatial atlas with an incredible breadth of molecular profiles across the developing lung in normal and disease states. Specifically, we will accomplish our goal of an integrated molecular atlas of lung development through the following aims: (1) Spatial imaging for a molecular atlas of the human lung in normal and diseased states, (2) Cell-specific omics for a molecular atlas of the human lung in normal and diseased states, and (3) Managing data to facilitate collaboration and data integration. Overall, these aims will create unprecedented multi-scale browsable quantitative three-dimensional “Google Maps” of proteins, lipids, and metabolites across the developing lung, providing for many novel insights toward understanding both normal human lung biology and disease pathogenesis. The novel imaging approaches and the suite of integrated pan-omics capabilities (i.e. proteomics, phosphoproteomics, activity-based proteomics, lipidomics and metabolomics) developed and available in a single laboratory at PNNL represents a unique strength of the Research Center.

PROJECT NARRATIVE High resolution molecular maps of human organs hold great promise for transforming our understanding of how organs function in health and disease states. The insights obtained from generating molecular maps of the lung will have significant impact in understanding diseases of significant burden associated with these organs, in particular rare lung diseases in children such as Bronchopulmonary Dysplasia (BPD), congenital diaphragmatic hernia, surfactant protein deficiencies, childhood interstitial lung diseases, pediatric pulmonary hypertension, and cystic fibrosis.