GSE163572
Targeting In Vivo Metabolic Vulnerabilities of Th2 and Th17 Cells Reduces Airway Inflammation
T cells promote inflammation in asthmatic patients and both Th2 and Th17 CD4 T cells have been implicated in severe forms of the disease. The metabolic phenotypes and dependencies of these cells, however, remain poorly understood in the regulation of airway inflammation. Here we show the bronchoalveolar lavage fluid of asthmatic patients had markers of elevated glucose and glutamine metabolism. Further, peripheral blood T cells of asthmatics had broadly elevated expression of metabolic proteins when analyzed by mass cytometry compared to healthy controls. Therefore, we hypothesized that glucose and glutamine metabolism were important in allergic airway inflammation. We tested this hypothesis in two murine models of airway inflammation. T cells from lungs of mice sensitized with Alternaria alternata extract displayed genetic signatures for elevated glucose and oxidative metabolism by single cell RNAseq.  This result was most pronounced when protein levels were measured in IL17 producing cells and was recapitulated when airway inflammation was induced with House Dust Mite plus lipopolysaccharide (HDM+LPS), a model which led to abundant IL4 and IL17 producing T cells. Importantly, inhibitors of the glucose transporter 1 (Glut1) or glutaminase (GLS) in vivo attenuated HDM+LPS eosinophilia, T cell cytokine production, and airway hyperresponsiveness as well as augmented the immunosuppressive properties of dexamethasone. These data show that T cells induce markers to support metabolism in vivo in airway inflammation and that this correlates with inflammatory cytokine production. Targeting metabolic pathways may provide a new direction to protect from disease and enhance the effectiveness of steroid therapy. Overall design: Adult mice were administered Alt Ext Ext (7.5ug) or vehicle control (PBS) on days 0, 3, 6. Lung were harvested on day 7 and digested using DNAse (0.1%) and collagenase (2mg/ml) for 30 minutes at 370C. Single cell lung suspensions were enriched for CD45+ cell using Miltenyi  microbeads (Catalog #130-052-301) using manufacturer's protocol. CD45+ cells were further purified by cell sorting and then loaded onto 10X Genomics Chromium Controller for single cell RNA sequencing.