(705.2) Targeting ADAM-17 to Alleviate Cytokine Storm Associated with COVID-19

Poster Board Number: B183

Vitoria Mattos Pereira (University of Wyoming), Suyasha Pradhanang (University of Wyoming), Amit Thakar (University of Wyoming, Washington State University), John Mansell (WyoBioTech), Sreejayan Nair (University of Wyoming)

Background: Acute respiratory distress syndrome (ARDS) is the primary cause of mortality in critically ill subjects infected with the novel coronavirus (severe acute respiratory syndrome-coronavirus-2, or SARS-CoV-2). Given the elevated levels of circulating proinflammatory cytokines such as tumor necrosis factors (TNF)-⍺ and interleukin (IL)-6, it has been postulated that a “cytokine storm” is responsible for the COVID-19 ARDS. A critical step in cytokine signaling is the shedding of membrane-anchored cytokines and their receptors. 

Objective: To determine the role of the metalloprotease/shreddase ADAM-17 (A disinterring and metalloproteinase 17) in COVID-19- mediated cytokine release from lung epithelial cells. We hypothesized pharmacological inhibition of ADAM-17 attenuated COVID-19-induced release of TNF-α in lung epithelial cells. Materials and

Methods: Human primary bronchial/tracheal epithelial cells (ATCC®, PS-300-010) were cultured to sub-confluence and challenged with COVID-19 Spike Coronavirus pseudovirus (SARS-COV-2 pseudo-virus, MyBioSource), and phorbol-12-myristate-13-acetate (PMA, Sigma), which was used as a positive control in the presence or absence of the ADAM-17 inhibitor TNF-α Protease Inhibitor-0 (TAPI-0, Sigma). TNF-α levels were determined in the cell media at different time points (2-12h) following the challenge using an ELISA assay. Data are represented as mean ± SEM, and statistical significance was determined by using the Student’s t-test.

Results: Treatment with both PMA (0.1 µM) and SARS-COV-2 pseudovirus (1x108 pseudoviral particles), resulted in a robust increase of TNF-α in the culture media 8 h post-stimulation (3243.75± 55.3 and 1278.6 ± 357 pg/mL respectively, p lt;0.05, n = 4).  Furthermore, treatment with TAPI-0 resulted in a concentration-dependent inhibition of the pseudovirus-induced TNF-α release by cultured bronchial epithelial cells, with a complete inhibition observed at 100 µM of TAPI-0.

Conclusion: Targeting ADAM-17 is a potential strategy to mitigated the cytokine storm associated with COVID-19-ARDS.

This work was supported in part by the WY Health and Bioscience Innovation Hub COVID Grant. Graduate student stipend was supported by a grant from the National Institutes of Health (P20GM103432).