Novel Pathway of Adenosine Generation in the Lungs from NAD⁺: Relevance to Allergic Airway Disease

Adenosine and uric acid (UA) play a pivotal role in lung diseases such as asthma and chronic obstructive pulmonary disease (COPD). In the present experiments, we measured adenosine synthesis from nicotinamide adenine dinucleotide (NAD⁺) in membranes prepared from wild type (WT) and CD38 knockout (CD38KO) mouse lungs, from cultured airway smooth muscle and epithelial cells, and in bronchoalveolar lavage fluid after airway challenge with epidemiologically relevant allergens. Adenosine was determined using an enzymatically coupled assay that produces ATP and is detected by luminescence. Uric acid was determined by ELISA. Exposure of cultured airway epithelial cells to Alternaria alternata extract caused significant nucleotide (NAD⁺ and ATP) release in the culture media. The addition of NAD⁺ to membranes prepared from WT mice resulted in faster generation of adenosine compared to membranes from CD38KO mice. Formation of adenosine from NAD⁺ affected UA and ATP concentrations, its main downstream molecules. Furthermore, NAD⁺ and adenosine concentrations in the bronchoalveolar lavage fluid decreased significantly following airway challenge with house-dust mite extract in WT but not in CD38KO mice. Thus, NAD⁺ is a significant source of adenosine and UA in the airways in mouse models of allergic airway disease, and the capacity for their generation from NAD⁺ is augmented by CD38, a major NADase with high affinity for NAD⁺. This novel non-canonical NAD⁺-adenosine-UA pathway that is triggered by allergens has not been previously described in the airways.