Rationale: SARS-CoV-2 gains entrance to airway epithelial cells (AECs) through binding of the viral spike protein to the angiotensin-converting enzyme 2 (ACE2) on the cell surface. However, ACE2 also converts angiotensin II into angiotensin-(1-7) and counterbalances the renin-angiotensin-aldosterone system, with resultant protective effects in the cardiovascular system. Some data suggest that two common antihypertension medications (angiotensin II receptor antagonists, ARBs; and angiotensin-converting-enzyme inhibitors, ACEIs) may increase ACE2 expression in heart and kidney cells, fueling debate about how these widely used medications may modulate SARS-CoV-2 infectivity and risk of COVID-19. Aim: Determine whether exposure of bronchial AECs to the ARB losartan or the ACEI captopril modulate expression of ACE2 by AECs, SARS CoV2 replication, or expression of proinflammatory cytokines and type I and III interferon (IFN) responses. Methods: Primary bronchial AECs from children and adults ( n = 19; Ages 8–75 yrs) were differentiated ex vivo at an air-liquid interface to generate organotypic cultures. Cultures were treated with captopril (1 μM) or losartan (2 μM) with culture media changes starting 72 h before infection with SARS-CoV-2. In a biosafety level 3 (BSL-3) facility, cultures were infected with SARS-CoV-2 isolate USA-WA1/2020 at a multiplicity of infection (MOI) of 0.5. At 96 h following infection, RNA and protein were isolated. SARS-CoV-2 replication in cultures was assessed with quantitative PCR (qPCR). ACE2, IL-6, IL-1B, IFNB1 , and IFNL2 expression were assessed by qPCR. Results: Neither captopril nor losartan treatment significantly changed ACE2, IL-6, IL-1B, IFNB1 , or IFNL2 expression by AECs as compared to SARS-CoV-2 infected AEC cultures without captopril or losartan treatment. At 96 h following infection, SARS-CoV-2 copy number/ng RNA was not significantly different between untreated AEC cultures, cultures treated with captopril, or cultures treated with losartan. Conclusion: These findings suggest that at the level of the airway epithelium neither the ACEI captopril or ARB losartan significantly modify expression of the SARS-CoV-2 entry factor ACE2, nor does either medication increase replication SARS-CoV-2 replication. This ex vivo data is reassuring and is consistent with evolving clinical data suggesting ACEIs and ARBs do not increase the risk for poor prognosis with COVID-19 and may actually reduce the risk of COVID-19 disease.
【저자키워드】 ACE2, airway, epithelium, losartan, Captopril, SARS—CoV—2, angiotensin-converting eitzyme inhibitors, angiotensin II (A II) receptor antagonists, 【초록키워드】 Treatment, SARS-CoV-2, IL-6, children, Infection, interferon, risk, cardiovascular system, angiotensin-converting enzyme 2, inhibitors, RNA, kidney, Replication, ARB, Protein, Culture, cells, qPCR, IFN, receptor, medication, ACE2 expression, disease, expression, change, quantitative PCR, SARS-CoV-2 replication, protective effect, binding, ACEi, Air-liquid interface, airway epithelium, ACEIs, culture media, Ex vivo, multiplicity of infection, viral spike protein, Proinflammatory cytokine, Clinical data, type I, poor prognosis, antagonists, cultures, IL-1B, USA-WA1/2020, IFNB1, responses, risk of COVID-19, Cell, Biosafety Level 3, BSL-3, MOI, were infected, significantly, generate, treated, modulate, changed, reduce, modify, AEC, airway epithelial cell, convert, IFNL2, infection with SARS-CoV-2, not significantly different, the SARS-CoV-2, were assessed, with COVID-19, 【제목키워드】 SARS-CoV-2, Angiotensin II, antagonist, Effect,