Introduction: SARS-CoV-2, the cause of COVID-19, belongs to the β-coronavirus subgroup that includes SARS-CoV-1 and MERS. SARS-CoV-2 utilizes the ACE2 receptor/TMPRSS2 protease complex expressed on the apical surfaces of respiratory epithelial cells as an entry mechanism. Removal of infectious agents from the airways is accomplished by mucociliary clearance (MCC), which requires a delicate balance between secretion and reabsorption of fluids. Ion channels such as the epithelial sodium channel (ENaC), the cystic fibrosis transmembrane conductance regulator (CFTR), and the calcium-activated chloride channel, TMEM16A, play key roles in regulating airway surface liquid homeostasis. SARS-CoV-2 encodes 4 major structural proteins: envelope (E), spike, membrane, and nucleocapsid proteins, which are required to produce a complete viral particle. CoV E proteins oligomerize to form viroporin channels that have been suggested to conduct cations and therefore may also affect airway hydration. Objectives: To elucidate the impact of SARS-CoV-2 on ion transport in airway epithelia and to determine whether the SARS-CoV E protein can be used as a novel and effective target to treat COVID-19 as well as future SARS outbreaks. Methods: We performed viral infections of primary airway epithelial cultures with CoV or CoV E. Because SARS-CoV-2 research requires BL3 facilities, the human CoV, NL63, which causes minor cold-like symptoms but shares the ACE2 entry mechanism with SARS-CoV-2, was used as a surrogate. We conducted measurements in Ussing chambers to determine ion channel function of CFTR, ENaC, and TMEM16A in the presence of CoV or SARS-CoV-2 E. In addition, we studied intracellular localization of CFTR, ENaC and SARS-CoV-2 E in primary airway cultures by immunofluorescence microscopy, and analyzed protein processing and expression by Western blotting. Furthermore, CoV E peptide studies in planar lipid bilayers by single-channel measurements were performed by fusion of membrane vesicles from HEK-293 expressing SARS-CoV-2 E or utilizing artificial lipid vesicles with reconstituted SARS-CoV-2 E peptides and conducting single-channel measurements. Results: CoV infection substantially affected ion channel function in primary airway epithelia with a significant reduction in ENaC activity, while function of CFTR and TMEM16A, similar to responses observed after airway inflammation (1), were enhanced. Furthermore, we demonstrated that SARS-CoV-2 E conducts cations, such as Na+ and K+ and notably, that hexamethylene amiloride (HMA) inhibits CoV E channels and reduces the titer of CoV. Conclusions: CFTR and ENaC were recently shown to be localized to secretory cells (2), where they regulate airway hydration required for MCC. Proper ENaC function requires proteolytic cleavage by the serine protease furin (3). We are currently exploring whether expression of viral spike protein, which contains the same furin cleavage sequence as ENaC, could interfere with ENaC processing. The most likely host-infection sequence for SARS-CoV-2 begins with infection of ciliated cells in the nose, followed by viral replication and shedding. Subsequent spreading of viral particles from the nasal cavity by inhalation results in infection of bronchiolar ciliated cells and, ultimately, alveolar cells (4,5). Understanding the role of CoV-2 E may be key to developing therapies to inhibit viral propagation in nasal and bronchial epithelia, before SARS-CoV-2 spreads to alveoli.