Project-002

Project 2 Summary Therapies for asthma that target type-2 (T2) inflammation are ineffective in many patients with asthma in part due to the increasing number of individuals with non-T2 asthma who have a suboptimal response to standard therapy with inhaled corticosteroids or biologics targeting T2 cytokines. Millions of viral-triggered asthma exacerbations occur annually in this group of patients in the U.S. leading to significant morbidity and cost to the health care system, due in part to the lack of effective treatments for non-T2 asthma for children and adults. The clinical features and airway inflammatory cell infiltrate patterns of non-T2 asthma have been characterized in detail, but the specific role of the airway epithelium in coordinating immune cell responses and influencing exacerbations, lung function, and persistent inflammation in donors with non-T2 asthma remains poorly characterized. Our preliminary studies demonstrate that airway epithelial cells (AECs) from non-T2 asthmatic children exhibit more robust type I and III interferon (IFN) responses and greater “non-T2 cytokine” (IL-1β, TNF-α, IL-17C, and GCSF) production following RV infection as compared to AECs from T2-high donors. In Project 2, we will use bronchial AECs from well characterized cohorts of children and adults with and without asthma to conduct mechanistic ex vivo experiments using organotypic models, to test our central hypothesis that in non-T2 asthma RV infection triggers excessive epithelial production of non-T2 cytokines and/or excessive IFN responses that activate the NLRP3 inflammasome, leading to neutrophilic inflammation, airway remodeling, and lung function decline through interactions with Th17 cells and/or lung macrophages. We will extend our ex vivo AEC model to include coculture studies of AECs with both Th17 and lung macrophages, and specifically assess bidirectional interactions utilizing bulk and single cell transcriptomics to assess epithelial-dependent neutrophil activation. We will also utilize cutting edge bioinformatics techniques to assess the genomic and epigenetic alterations in the epithelium that drive non-T2 inflammation as well as relevant in vivo modeling of HRV infection in the setting of non-T2 inflammation.