Role of KLF6 in Lung Development - SUMMARY/ABSTRACT Despite the lungs’ crucial role in health and disease, significant gaps remain in our understanding of the mechanisms that govern the development of the normal, healthy lung. Adressing these knowledge gaps will inform therapeutic options for disease and is vital for ensuring long-term respiratory health in both children and adults. As a Pediatric Pulmonologist and physician-scientist, my long-term goal is to identify and validate new therapeutic targets to improve the care of children with respiratory diseases. Toward this effort, our group recently uncovered a novel regulator of lung development, Krüppel-like factor-6 (KLF6). In mice, my preliminary data show that inactivation of Klf6 in the developing lung epithelium leads to a lethal lung growth defect, and inactivation of Klf6 in alveolar epithelial type 1 (AT1) cells just after birth leads to a reduction in lineage traced AT1s. Taken together, this suggests that KLF6 is required in distinct biological processes in prenatal lung growth versus postnatal alveolar formation. By leveraging single-cell profiling, multimodal data analysis and integration, and mouse genetic approaches, my overall objective is to 1) define the roles of KLF6 in branching morphogenesis and postnatal alveolar formation, and 2) define the KLF6-regulated target genes and signaling pathways. My central hypothesis is that KLF6 is a critical promoter of lung growth, airway branching, and alveolar epithelial cell formation. By delineating the role and mechanism of a novel transcriptional regulator in lung development, my findings will contribute to new strategies to address childhood lung diseases such as bronchopulmonary dysplasia – a chronic lung disease in premature infants with lifelong impacts on lung health. The central hypothesis will be tested by pursuing two specific aims: 1) Delineate the mechanism of KLF6 function in lung growth during prenatal branching morphogenesis; and 2) Determine the mechanism of KLF6 function in alveolar epithelium formation during postnatal alveologenesis. Mouse Cre lines will be used to define the cellular mechanisms underlying the phenotypes observed, and sequencing-based omics including state of the art 10X Multiome – simultaneous profiling of gene expression and chromatin accessibility from the same nucleus – will be used to precisely define cellular states and their regulation. The research proposed in this application is innovative, in the applicant’s opinion, because it will determine the role of a poorly studied transcription factor regulator, KLF6, in lung development. The proposed research is significant because it will provide the scientific premise for my future R01 proposals to advance our understanding of both normal lung development and disease pathogenesis.
