The role of extracellular histones in COVID-19.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread from China within 2 months to become a global pandemic. Infection can cause a diversity of symptoms ranging from asymptomatic to severe acute respiratory distress syndrome (ARDS) with an increased risk of vascular hyperpermeability, pulmonary inflammation, extensive lung damage and thrombosis. One of the host defense systems in coronavirus disease 2019 (COVID-19) is the formation of neutrophil extracellular traps (NETs). Numerous studies have revealed elevated levels of NET components, such as cell-free DNA (cfDNA), extracellular histones, neutrophil elastase (NE) and myeloperoxidase (MPO), in plasma, serum and tracheal aspirates of severe COVID-19 patients. Extracellular histones, a major component of NETs, are clinically very relevant since they represent promising biomarkers and drug targets given that several studies have identified histones as key mediators in the onset and progression of various diseases, including COVID-19. However, the role of extracellular histones in COVID-19 per se remains relatively under-explored. Histones are nuclear proteins that can be released into the extracellular space via apoptosis, necrosis or NET formation and are then regarded as cytotoxic damage-associated molecular patterns (DAMPs) that have the potential to damage tissues and impair organ function. This review will highlight the mechanisms of extracellular histone-mediated cytotoxicity and focus on the role that histones play in COVID-19. Thereby this paper facilitates a bench-to-bedside view of extracellular histone-mediated cytotoxicity, its role in COVID-19, and histones as potential drug targets and biomarkers for future theranostics in the clinical treatment of COVID-19 patients. This article is protected by copyright. All rights reserved.

Impact of Dutch COVID-19 restrictive policy measures on physical activity behavior and identification of correlates of physical activity changes: a cohort study.

BACKGROUND: Identification of characteristics of individuals that are related to decreases in physical activity (PA) levels during lockdown is needed to develop targeted-interventions. This study aims to evaluate changes in domain-specific (i.e. leisure time, transportation, occupational, and household) and total PA due to the Dutch COVID-19 lockdown, which started on March 15 2020. Furthermore, we aim to identify demographic, health-related, and psychological correlates of these changes. METHODS: Individuals who participated in the Nijmegen Exercise Study during 2017-2019 were invited to this study, which was conducted between April 16 and May 12 2020. Participant characteristics (i.e. age, sex, body mass index (BMI), marital status, education, household composition, and occupation status), living environment (i.e. housing type and degree of urbanization), psychological characteristics (i.e. resilience, outcome expectations, vitality, and mental health), and medical history were collected via an online questionnaire. Short Questionnaire to Assess Health-enhancing physical activity was used to assess PA behavior before and during lockdown. Wilcoxon signed-rank test was used to compare PA levels, in metabolic equivalent of task (MET)-minutes per week (min/wk), before and during lockdown. Multivariable linear regression analyses were performed to examine correlates of PA changes. RESULTS: 4033 participants (57% male; 59 ± 13 years) were included. PA decreased significantly during lockdown with mean ± SD changes of 393 ± 2735 MET-min/wk for total, 133 ± 785 MET-min/wk for transportation, 137 ± 1469 MET-min/wk for occupation, and 136 ± 1942 MET-min/wk for leisure time PA. Household PA did not change significantly. Unemployment, COVID-19-related occupational changes, higher BMI, and living in an apartment or semi-detached/terraced house were significantly related to larger decreases in total and domain-specific PA. Higher vitality was related to smaller decreases in total and domain-specific PA. Higher age was significantly associated with a larger decrease in leisure time PA. Lower education was associated with smaller decreases in transportation and occupational PA compared to higher education. CONCLUSION: PA levels significantly reduced during lockdown compared to before lockdown. Declines were observed during transportation and occupation, but were not compensated by an increase in leisure time PA. We identified subgroups that were more susceptible to reductions in domain-specific or total PA levels and should therefore be encouraged to increase their PA levels during lockdown.

Changes in Physical Activity and Sedentary Behaviour in Cardiovascular Disease Patients during the COVID-19 Lockdown.

The COVID-19 lockdown has been associated with physical inactivity. We prospectively evaluated changes in moderate-to-vigorous physical activity (MVPA) and sedentary time (ST) among 1565 cardiovascular disease (CVD) patients using validated questionnaires at 5 weeks after lockdown initiation (i.e., baseline, April 2020) and at every 4 subsequent weeks, until July 2020. Multivariate mixed model analyses were performed to identify whether age, sex, CVD-subtype, lockdown adherence and mental health factors impacted changes in physical (in)activity. Patients were 67 (interquartile range: 60-73) years and primarily diagnosed with coronary artery disease. Time spent in MVPA was 143 min/day (95% confidence interval (CI) 137; 148) at baseline. Female sex, heart-failure, fear of COVID-19 infection and limited possibilities for physical activity were independently associated with lower levels of MVPA across time. After adjusting for confounders, overall MVPA did not change. ST was 567 (95% CI 555; 578) min/day at baseline. Lack of social contact, limited possibilities for physical activity and younger age were independently associated with higher levels of ST. After adjusting for confounders, ST progressively increased following 8 (Δ+19.7 (95% CI 0.4; 39.0)) and 12 weeks (Δ+25.2 (95% CI 5.4; 47.1) min/day) of lockdown. Despite a phased relaxation of the lockdown, CVD patients progressively increased ST and reported no change in MVPA. This highlights the need to target physical inactivity during and beyond the COVID-19 pandemic.

Histone H3 Cleavage in Severe COVID-19 ICU Patients.

The severity of coronavirus disease 19 (COVID-19) is associated with neutrophil extracellular trap (NET) formation. During NET formation, cytotoxic extracellular histones are released, the presence of which is linked to the initiation and progression of several acute inflammatory diseases. Here we study the presence and evolution of extracellular histone H3 and several other neutrophil-related molecules and damage-associated molecular patterns (DAMPs) in the plasma of 117 COVID-19-positive ICU patients. We demonstrate that at ICU admission the levels of histone H3, MPO, and DNA-MPO complex were all significantly increased in COVID-19-positive patients compared to control samples. Furthermore, in a subset of 54 patients, the levels of each marker remained increased after 4+ days compared to admission. Histone H3 was found in 28% of the patients on admission to the ICU and in 50% of the patients during their stay at the ICU. Notably, in 47% of histone-positive patients, we observed proteolysis of histone in their plasma. The overall presence of histone H3 during ICU stay was associated with thromboembolic events and secondary infection, and non-cleaved histone H3 was associated with the need for vasoactive treatment, invasive ventilation, and the development of acute kidney injury. Our data support the validity of treatments that aim to reduce NET formation and additionally underscore that more targeted therapies focused on the neutralization of histones should be considered as treatment options for severe COVID-19 patients.

Evolution of NETosis markers and DAMPs have prognostic value in critically ill COVID-19 patients.

Coronavirus disease 19 (COVID-19) presents with disease severities of varying degree. In its most severe form, infection may lead to respiratory failure and multi-organ dysfunction. Here we study the levels and evolution of the damage associated molecular patterns (DAMPS) cell free DNA (cfDNA), extracellular histone H3 (H3) and neutrophil elastase (NE), and the immune modulators GAS6 and AXL in relation to clinical parameters, ICU scoring systems and mortality in patients (n = 100) with severe COVID-19. cfDNA, H3, NE, GAS6 and AXL were increased in COVID-19 patients compared to controls. These measures associated with occurrence of clinical events and intensive care unit acquired weakness (ICUAW). cfDNA and GAS6 decreased in time in patients surviving to 30 days post ICU admission. A decrease of 27.2 ng/mL cfDNA during ICU stay associated with patient survival, whereas levels of GAS6 decreasing more than 4.0 ng/mL associated with survival. The presence of H3 in plasma was a common feature of COVID-19 patients, detected in 38% of the patients at ICU admission. NETosis markers cfDNA, H3 and NE correlated well with parameters of tissue damage and neutrophil counts. Furthermore, cfDNA correlated with lowest p/f ratio and a lowering in cfDNA was observed in patients with ventilator-free days.