Proteome Profiling of Recombinant DNase Therapy in Reducing NETs and Aiding Recovery in COVID-19 Patients

Abstract

Severe COVID-19 can result in pneumonia and acute respiratory failure. Accumulation of mucus in the airways is a hall mark of the disease and can result in hypoxemia. Here, we show that quantitative proteome analysis of the sputum from severe COVID-19 patients reveal high levels of neutrophil extracellular trap(s) (NETs) components, which was confirmed by microscopy. Extracellular DNA from excessive NET formation can increase sputum viscosity and can lead to acute respiratory distress syndrome (ARDS). Recombinant human DNase (rhDNase/Pulmozyme) has been shown to be beneficial in reducing sputum viscosity and improve lung function. We treated 5 COVID-19 patients presenting acute symptoms with clinically approved aerosolized Pulmozyme. No adverse reactions to the drug were seen, and improved oxygen saturation and recovery in all severely ill COVID-19 patients was observed after therapy. Immunofluorescence and proteome analysis of sputum and blood plasma samples after treatment revealed a marked reduction of NETs and a set of statistically significant proteome changes that indicate reduction of haemorrhage, plasma leakage and inflammation in the airways, and reduced systemic inflammatory state in the blood plasma of patients. Taken together, the results indicate that NETs contribute to acute respiratory failure in COVID-19 and that degrading NETs may reduce dependency on external high flow oxygen therapy in patients. Targeting NETs using rhDNase may have significant therapeutic implications in COVID-19 disease and warrants further studies.

Neutrophil Extracellular Trap Formation Potential Correlates with Lung Disease Severity in COVID-19 Patients

“…Further investigation is needed to determine the source(s) and clinical implications of circulating DNA fragments in COVID-19 patients to better target them therapeutically. Broad targeting of DNA fragments with DNase has demonstrated promising results [ 53 , 54 ] in pilot studies, but there is some evidence that the timing and route of treatment may be critical for positive outcomes [ 55 ].…”

Pulmonary fibrosis from molecular mechanisms to therapeutic interventions: lessons from post-COVID-19 patients

“…Generally, the cytokine storm and ARDS have been demonstrated to be promoted by NETs that play the main role in the interaction between neutrophils and macrophages during the early acute phase of acute lung injury [199] . Indeed, NETs contribute to acute respiratory failure in COVID-19 and the use of tools able to degrade NETs may reduce inflammation and improved oxygenation in patients [200] . However, the mechanisms regulating NET formation are currently unclear as is its implication in COVID-19-induced PF.…”

A Review of the Neutrophil Extracellular Traps (NETs) from Cow, Sheep and Goat Models

“…Phytochemicals and probiotics [99,105,128,129] are being studied to augment innate immunity against microbes and gastrointestinal parasites to address concerns regarding antibiotic resistance. These include extracts such as: garlic, neem, wormwood, tobacco, cowpea [87,130,131] and Sericea lespedeza [132].…”

“…In cattle sheep and goats, changes in NET markers and the proteome are associated with disease of concern to animal health and production [72]. Increased definition of the proteomes, associated post-translational modifications, and markers of NETosis in sheep cattle and goats will contribute to the development of therapeutics for inhibition of excess NETs formation to ameliorate pathologies associated with diseases [101,129,152,158].…”