PROJECT SUMMARY / ABSTRACT Asthma is a complex, inflammatory disease of the lung whose prevalence continues to rise in the United States, with one in twelve individuals afflicted. Although inhaled corticosteroids and β2-agonists have been the mainstay of treatment regimens, these therapies have varying efficacy in a heterogeneous patient population and are recommended based on symptom alleviation and control, rather than targeting pathogenic mechanisms. Individuals most commonly suffer from allergic asthma, characterized by hypersensitivity of airway epithelial cells to inhaled allergens, leading to a type 2 adaptive immune response. Immune cells release type 2 cytokines, which trigger airway epithelial cells through members of the STAT (signal transducer and activator of transcription) family, causing many of the clinicopathologic features of asthma. Thus, airway epithelial cells represent a critical interface in the development of asthma. Despite this, the regulation of downstream events in airway epithelium following type 2 cytokine stimulation is not well understood. While many studies have elucidated the role of the coding genome in the development of allergic asthma, regulation by the non-coding genome is not well described. Long noncoding RNAs (lncRNAs) have increasingly been identified as important functional molecules in regulating an immune response and cellular identity. We discovered the lncRNA WFDC21P is highly expressed following IL-13 stimulation in primary human bronchial epithelial cells. WFDC21P was previously found to regulate dendritic cell differentiation and function, however its role in airway epithelial cells has not yet been studied. Our preliminary data suggest WFDC21P directs differentiation of airway epithelial cells via regulating the level of STAT3 phosphorylation. To test this hypothesis, we propose a combination of advanced genomic and molecular tools to study the effects of WFDC21P on a genomic, cellular, and molecular level. First, we will determine the cell-type specificity of WFDC21P following IL-13 induction and determine its cellular and molecular effects on airway epithelial morphology and STAT signaling. These studies will be performed via single-cell RNA sequencing, histology, immunofluorescence, and flow cytometry. Subsequently, we will assess the effects of WFDC21P on the genome using RNA interference, ATAC-sequencing, and RNA- sequencing followed by a series of computational analyses. Completion of these studies will provide key biological insight into the precise downstream regulation of STAT signaling in airway epithelial cells and how lncRNAs contribute to airway biology and the type 2 immune response. These studies will thus provide novel pathways and regulatory targets to be used for future therapeutics in the treatment of asthma.