TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS最新文献

{"title":"COVID-19 and Obstructive Lung Disease: Are COPD and Asthma Risk Factors for Severe COVID-19? Evaluating the Data from the Largest Health System in New York State","authors":"Jasmine Schwartz, B. Birnbaum, M. Ballenberger, Omar Mahmoud, B. Mina","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1276","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1276","url":null,"abstract":"RATIONALE: An analysis of data collected between March 1, 2020 through July 1, 2020 from the largest health system in New York sought to investigate the association between COVID-19 and obstructive lung disease including asthma and chronic obstructive pulmonary disease (COPD). Methods: A total of 21,865 patients were included in the analysis, 2,518 had obstructive lung disease and COVID-19. There were 1,370 [HA1] patients who had asthma and COVID-19, and 847 patients had COPD and COVID-19. There were 301 patients who had overlapping asthma and COPD diagnoses. There were 19,347 patients who had a diagnosis of COVID-19 without asthma or COPD. Logistic regression analysis of the data was performed. Results: Analysis of our data showed COPD patients with COVID-19 were more likely to be intubated as compared to the control group (OR 1.346 95% CI 1.061-1.707 p=0.0095). Additionally, there was no association between COPD and mortality in patients with COVID-19 (OR 0.845, 95% CI: 0.685-1.042 p=0.1436). There was no association between asthma and COVID-19 and intubation (OR 1.175 95% CI 0.936-1.475 p=0.2193). There was no statistical association between asthma and mortality in patients with COVID-19 (OR 0.849 95% CI 0.669-1.079 p=0.2455). However, patients with asthma were at a decreased risk of dying from COVID-19 compared to COPD patients with COVID-19 even after controlling for comorbid conditions (OR 0.718, 95% CI 0.526-0.979 p=0.0329). Patients presenting with asthma exacerbation or COPD exacerbation in the setting of COVID-19 infection were not at increased risk of death as compared to COPD or asthma patients with COVID-19 (Asthma exacerbation OR 1.042, 95% CI 0.62-1.73 p=0.8796), (COPD exacerbation OR 0.95 95% CI 0.67-1.36 p=0.762). Conclusion: Our finding with regards to intubation may be explained by an aggressive approach early on in the pandemic towards intubation especially in patients that may have been perceived as high-risk patients' due to underlying lung disease. Our data also showed no association between COPD and death in COVID-19 and asthma and mortality in COVID-19-meaning COPD and Asthma were not risk factors for death in our patient population. As to why asthma patients were at decreased odds of dying compared to COPD patients may be related to the different pathophysiological mechanisms of asthma as compared to COPD or a small protective effect of inhaled corticosteroids. Lastly, presenting with an asthma exacerbation or COPD exacerbation in the setting of COVID-19 infection did not increase your odds of mortality.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121045796","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of ACE1 (Angiotensin Converting Enzyme 1) Polymorphism Rs1799752 on Protein Levels of ACE2, the SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) Entry Receptor, in Alveolar Lung Epithelium","authors":"M. Jacobs, L. Lahousse, H. V. Eeckhoutte, S. Wijnant, J. Delanghe, G. Brusselle, K. Bracke","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1272","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1272","url":null,"abstract":"Rationale One of the striking features of the Coronavirus Disease 19 (COVID-19) pandemic is the considerable variation in disease presentation and severity amongst patients, ethnic groups, and countries. Genetic factors could contribute to this variation. The Angiotensin-Converting Enzyme 1 (ACE1) gene is characterized by a genetic deletion/insertion (D/I) of an alu repeat and this polymorphism (rs1799752) shows an important geographical variation. Recently, we showed that COVID-19 incidence was inversely correlated to the presence of the ACE1 D-allele frequency. Also, a significant correlation between COVID-19 related mortality and the prevalence of the D-allele was observed. ACE1 is a homologue of ACE2, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) host cell entry receptor. Since ACE1 and ACE2 levels are strongly regulated by common genetic variants in their genes, ACE2 levels may also be influenced by this polymorphism. Therefore, this genetic D/I in ACE1 might influence infectivity and pathogenicity of SARS-CoV-2. Methods ACE2 protein expression was visualized by immunohistochemistry on lung tissue sections from 67 subjects and quantified in alveolar epithelium. DNA was extracted from lung tissue and the prevalence of the three genotypes of rs1799752 (II, DI, DD) in ACE1 was determined using Taqman® SNP Genotyping assays. Results Significantly higher levels of ACE2 protein in alveolar lung epithelium were observed when subjects were homozygous for the insertion (II), compared to subjects homozygous for the deletion (DD). Importantly, this correlation remained significant, even after adjusting for age, sex, BMI, COPD, smoking, and diabetes. Therefore, homozygosity of the insertion (II) is an independent determinant for increased lung ACE2 levels and is not driven by higher frequencies of the insertion in certain patient groups. Conclusion In conclusion, we suggest a genetic D/I polymorphism in ACE1 to correlate with alveolar protein expression of ACE2, the SARS-CoV-2 entry receptor, thereby potentially affecting infectivity and pathogenicity of SARS-CoV-2. The geographical variance in the ACE1 II genotype might contribute to the varying prevalence, morbidity, and mortality due to COVID-19.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"103 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123251279","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of Angiotensin Converting Enzyme Inhibitor or Angiotensin II Receptor Blocker Treatment of Differentiated Primary Airway Epithelial Cells on Ace2 Expression and SarsCoV2 Replication","authors":"L. M. Rich, K. Barrow, E. Vanderwall, S. R. Reeves, M. White, W. E. Harrington, J. Debley","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1284","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1284","url":null,"abstract":"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, ACE-2 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 blockers, ARBs;and angiotensin-converting-enzyme inhibitors, AECi) may increase ACE-2 expression in heart and kidney cells, fueling debate about how these widely used medications may modulate SARS-CoV2 infectivity and risk of COVID-19. Aim: Determine whether exposure of bronchial AECs to the ARB losartan or the ACEi captopril modulate expression of ACE-2 by AECs and/or SARS CoV2 replication. Methods: Primary bronchial AECs from children and adults (n=18;ages 8-75 yrs.) were differentiated ex vivo at an air-liquid interface to generate organotypic cultures. Cultures were treated with captopril (1μM) or losartan (2μM) added to basolateral media with each culture media change starting 72 hrs. 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 hrs. following infection, RNA and protein were isolated from cultures. SARS-CoV-2 replication in cultures was assessed with quantitative PCR, and quantified as viral copy number/ng RNA. ACE-2 expression was assessed by qPCR. Results: Neither captopril nor losartan treatment significantly changed AEC-2 expression by AECs as compared to untreated AEC cultures or SARS-CoV-2 infected AEC cultures without captopril or losartan treatment. At 96 hrs. following infection, SARS-CoV-2 copy number/ng RNA was not significantly different between untreated AEC cultures (median 1752, 95% CI 213 - 8599), cultures treated with captopril (median 448.6, 95% CI 160 - 3051), or cultures treated with lorsartan (median 640, 95% CI 127 - 1610;Kruskal-Wallis ANOVA p=0.4). 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 ACE-2, nor does either medication effect the replication of SARS-CoV-2. This ex vivo data is reassuring and is consistent with evolving clinical data suggesting ACEi and ARB medications do not increase the risk for poor prognosis with COVID-19, and may actually reduce the risk of COVID-19 disease.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"23 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128515557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Relationship Between Ace2 and Tmprr2 Expression by Differentiated Primary Bronchial Airway Epithelial Cells and SarsCoV2 Replication","authors":"E. Vanderwall, K. Barrow, L. M. Rich, S. R. Reeves, M. White, N. Sather, W. Harrington, J. Debley","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1281","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1281","url":null,"abstract":"RATIONALE: SARS-CoV-2 gains entrance to airway epithelial cells (AECs) via binding of the viral spike protein to the angiotensin-converting enzyme 2 (ACE2) on the cell surface, and the serine protease TMPRSS2 is thought to play an important role in facilitating SARS-CoV-2 entry by priming the spike protein. There is some data suggesting that ACE-2 expression by AECs is greater in adults than children, leading many to hypothesize that airway ACE-2 expression is a risk factor for SARS-CoV-2 replication and COVID-19 disease. Aim: Determine whether expression of ACE-2 and/or TMPRSS2 by bronchial AECs from children and adults is associated with SARS CoV2 replication. Methods: Primary bronchial AECs from children and adults (n=18;ages 8-75 yrs.) were differentiated ex vivo at an air-liquid interface to generate organotypic cultures. 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 hrs. following infection, RNA and protein were isolated from cultures. SARS-CoV-2 replication in cultures was assessed by PCR, and quantified as viral copy number/ng RNA. ACE-2 expression was assessed by qPCR in both SARS-CoV-2 infected AEC cultures and uninfected control cultures. In a subset of subjects (n=6), ACE-2 expression was measured in paired nasal and bronchial AEC cultures. Finally, we assessed the effect of apical treatment of AEC cultures with recombinant ACE-2 (rACE-2) prior to SARS-CoV-2 and once daily for 96rs. Results: In the primary bronchial AECs studied we observed marked between subject heterogeneity in ACE-2 expression (14-fold), TMPRSS2 expression (8-fold), and SARS-CoV-2 replication (range 167-89,040 copies/ng RNA). Baseline ACE-2 expression in uninfected AECs correlated with SARS-CoV-2 replication in infected AECs (Spearman r=0.6, p=0.02), whereas TMPRSS2 expression was not associated with viral replication (r=-0.2, p=0.5). In paired nasal and bronchial AEC cultures ACE-2 expression was strongly correlated (Pearson R2=0.66, p=0.05). Treatment of AECs with rACE-2 added apically immediately prior to infection and refreshed daily for 96 hrs. across a range of concentrations (0.1-1000 ng/mL rACE in 100μL of PBS;n=4 AEC primary lines) led to a marked reduction in SARS-CoV-2 replication (mean of 5040 viral copies/ng RNA in untreated AECs to 16 viral copies/ng RNA at 10ng/mL). Conclusion: Expression of ACE-2 by primary bronchial AECs from children and adults is heterogenous, and is associated with SARS-CoV-2 replication ex vivo. ACE-2 expression by AECs may partially explain the between subject variability in the risk and severity of COVID-19.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"57 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121735918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Impact of Age, Gender and Active Cancer in the Mortality and the Length of In-Patient Stay of Patients with COVID-19 at A UK District General Hospital","authors":"H. Ahmad, M. Butt, R. Ragatha, S. Naik, U. Ekeowa, P. Russell, K. Khan, A. Mirza, M. Anwar","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1286","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1286","url":null,"abstract":"RATIONALE: An age-related mortality risk has been discovered during the COVID-19 pandemic;the elderly being at greater risk. This could be explained by age-related impairment of immunity. Social factors such as congregate housing could also play a role[1]. Similarly, cancer patients have been identified as being high-risk for mortality. Thromboembolic events arising as a result of a cytokine storm has been theorised as a potential cause of death as illustrated in figure 1[2]. Previous studies have highlighted male sex as being another high-risk group for morbidity and mortality[2] . We aim to investigate the effects of age, gender and active cancer on the mortality rate and the length of in-patient stay in patients with COVID-19. Methods: A retrospective study of all in-patients aged ≥ 18 years with a confirmed diagnosis of COVID-19 during the first wave of the pandemic. Statistical analysis was performed using the chi squared and Mann-Whitney U test. Results: 445 COVID-19 positive patients were included in the study, of which 69 had active cancer, 329 were aged <65 years and 116 were aged > 65 years. The study contained 261 males and 263 females. Mortality in patients with active cancer was higher (70%) compared to those without active cancer (48%) (P=0.001). There was no significant difference in the number of patients who had an inpatient stay of >7 days between both groups. We also found that there was a higher mortality rate in patients aged > 65 years (61%) compared to those aged < 65 years (25%) (P<0.05), with a greater number of patients aged > 65 years staying >7 days in-hospital (63%) compared to those aged < 65 years (49%) (P=0.03). There were no significant differences in the mortality rates and the length of in-patient stays of >7 days between male and female patients. However, interestingly males had a greater intubation rate (14%) compared to females (6%) (P=0.025). Conclusion: Our study demonstrated increasing age and active cancer status to be linked to greater risk of mortality. Furthermore, males showed a more severe disease course as compared to females. This data should be considered when highlighting at risk groups and prioritising them for treatment and isolation. Figure 1-A potential mechanism of death in COVID-19 patients References: 1. Kang SJ et al Age-related Morbidity and Mortality among patients with COVID-19. Infect Chemother 2020;52(2):154-164 2. Curigliano G et al Cancer Patients and Risk of Mortality for COVID-19. Cancel Cell 2020;38(2):161-163.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"43 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133177694","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Severe Asthma Patients Experience and Satisfaction with Virtual Clinics in a Large Tertiary Care Hospital During COVID-19 Period","authors":"H. Al-Jahdali, Mohammad A Khan, A. Al-Harbi, R. Rajkumar, M. Al-Gamedi","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1271","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1271","url":null,"abstract":"Background: Enforced social distancing (i.e. lockdowns) greatly facilitated control of COVID-19. Whilst access to hospitals was restricted, outpatient care continued remotely. At our institute, the biologic therapy for severe asthma patients is administered on-site by specialist nurses who follow manufacturers' recommendations. Aim: The aim of this study was to determine the satisfaction of patients with severe asthma with telemedicine, and the impact of COVID-19 lockdown on their receipt of biologics and other treatments for asthma. Methods: A crosssectional survey of 58 patients with severe asthma scheduled to receive biologic therapy at our hospital during the lockdown was performed with ethical approval. Results: Fifty-four patients participated (F 37;mean age 46.7 years;response rate 93.1%). Meantime since diagnosis was 19.2 years (SD 11.5 years). All had been on biologic therapy Omalizumab (45), Mepolizumab (7), or Dupilumab (2) for over three months (mean 38.4 months ± SD 26.5 months). Fifty (92.6%) had telephone follow-up, 31 (57.4%) were satisfied with telemedicine, 45 (81.4%) agreed that biologic therapy improved their asthma, and 40 (74.1%) received scheduled biologic therapy. Of the 45 patients living in the city, nine did not receive biologic therapy, two cited the lockdown as the reason for this;two did not receive an appointment;two did not perceive any benefit;2 had other reasons. Five of the nine patients living outside the city did not receive biologic therapy, 3 because of the lockdown, and 1 for fear of acquiring COVID-19. Alarmingly, 16 (29.6%) suggested that they had insufficient medications, and 27 (50%) reported difficulty obtaining medications. Conclusions: Many patients were satisfied with telemedicine, so this could be used to deliver routine outpatient tertiary care post-pandemic. However, logistics around supplying medications, and biologics must be considered in plans preparing for the second wave of COVID-19.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"26 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133510049","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Phase IIa, Double-Blinded, Randomized Placebo-Controlled Trial of MAPKAPK2 Inhibition by ATI-450 in Treatment of Moderate-Severe COVID-19 Pneumonia","authors":"D. Polineni, M. Jandali, C. Streiler, U. Nazir, M. P. Jasahui, D. Morgan, H. Smith, L. Holets-Bondar, M. Markiewicz, J. He, E. Fruhauf, A. Godwin, S. Soper, D. Koestler, D. Gordon, M. Castro, G. Gan","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1287","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1287","url":null,"abstract":"RATIONALE: The novel coronavirus disease 2019 (COVID-19) is a rapidly spreading global viral pandemic with a high-risk of mortality in selected populations (i.e. elderly, immunocompromised, cardiopulmonary diseases). A hyperinflammatory state caused by excessive inflammatory cytokine production (i.e. TNFα, IL-1β, IL6, IL-8) has been attributed to the pathobiology of COVID-19-mediated acute respiratory distress syndrome, worsened lung fibrosis and increased mortality. Inflammation and inflammatory disorders signal primarily through the MAPK pathway. Activation of p38α is important for regulating inflammation, and aberrant p38α activation is associated in the pathobiology of diseases such as idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease and tissue fibrosis. The downstream target of p38α is the protein MAPKAPK2 (MK2) and is responsible for transcriptional production of pro-inflammatory cytokines also elevated in COVID-19 inflammation. Further evidence in other severe viral illnesses (i.e. Dengue, Influenza, CMV) show activation of p38-MK2 signaling axis for mediating inflammation. Potentially, COVID-19 mediated inflammatory cytokine production may signal through p38α-MK2 axis and MK2 pathway blockade may suppress unwanted inflammation. However, it remains unknown whether reducing the inflammatory state can improve COVID-19 outcomes particularly in those with pre-existing conditions. We hypothesized that blockading this pathway would reduce inflammatory cytokine burden and improve respiratory failure-free survival in moderate-severe COVID-19 infected patients. Methods: We designed an investigator-initiated trial (IND#:149790;ClinicalTrials.gov Identifier: NCT04481685) using an oral MK2 inhibitor (ATI-450, Aclaris Pharmaceuticals) in COVID-19. This single-center trial is a Phase IIa, doubleblinded, randomized placebo-controlled proof-of-concept study. COVID-19 positive hospitalized patients with pulmonary signs and symptoms of moderate-severe hypoxic respiratory distress were randomized to ATI-450 or placebo twice-daily for up to 14 days. Results: Study enrollment is completed (n=20 subjects;n=11 male;median age 63 years old). The primary endpoint of this trial is respiratory failure-free survival at 14 days. Secondary endpoints include: changes in WHO-Ordinal scale, additional respiratory and survival outcomes, biochemical assays of circulating cytokines, and safety endpoints. Given the incomplete knowledge of MK2 pathway blockade effects on immune cell function, this study will further explore immune cell characterization in COVID-19 patients treated with ATI-450 via immunophenotyping and 10X Genomics single-cell gene expression analysis. We surmise that myeloid cell activation following COVID-19 infection contributes to localized and systemic tissue injury and will examine the effect of MK2 pathway blockade on eliciting myeloid cell inflammatory activation-suppression. Conclusion: Analyses of the safety, efficacy, and biology","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134233078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cigarette Smoke Exposure Increases ACE-2 Expression and SARS-CoV-2 Infection Severity in Human and Ferret Airways and Induces Apoptotic Cell Injury In Vitro","authors":"S. S. Hussain, J. Tipper, S. Phillips, J. Campos-Gómez, Q. Li, K. Harrod, J. E. P. Lever, S. Rowe","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1277","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1277","url":null,"abstract":"RATIONALE: Cigarette smoking is associated with COVID-19 prevalence and severity, but the mechanistic basis for how smoking alters SARS-CoV-2 pathogenesis is unknown. A potential explanation is that smoking could alter expression of angiotensin converting enzyme-2 (ACE2), which functions as the cellular receptor and point of entry. Here we investigated the severity of SARS CoV-2 infection ex vivo and in vitro and using tissue samples and primary ferret and human airway epithelial cells with and without antecedent exposure to cigarette smoke. Methods: ACE2 expression measured by Quantitative PCR (Q-PCR) of ferret lungs exposed to 6 months of cigarette smoke, and findings validated by immunofluorescence (IF). Primary airway cells isolated from airways of ferrets or human non-smokers or COPD subjects were grown until terminally differentiated at air liquid interface. Cells were then exposed to cigarette smoke extract (CSE) or vehicle control, then infected with SARS CoV-2 (3 MOI) or mock control. Viral copy was measured by Q-PCR. Viral infection was quantified by foci forming assay (FFU/mL) using VeroE6 cells. Results: Ferret lungs following 6 months of smoke exposure had increased ACE2 as compared to air controls by Q-PCR (>1.5 fold, P<0.05, N=6), and IF staining. Higher ACE2 expression was also observed in normal ferret airway cells exposed to CSE (>1.5 fold, P<0.05, N=3), normal Human Bronchial Epithelial (HBE) cells exposed to CSE (>2 fold, P<0.001, N=4), and HBE cells from COPD donors as compared to healthy controls (>2 fold, P<0.001, N=4). When ferret airway cells were inoculated with SARS-CoV-2, intracellular viral load of SARS-CoV-2 was increased in CSE exposed cells as compared to vehicle controls (103-104 vehicle Vs CSE 105 106 copy/μL). Viral infection was also increased >2 fold (P<0.01, N=5). Likewise, CSE exposed (105-107 copy/μL, P<0.0001, N=4) and COPD (106-108 copy/μL, P<0.0001, N=4) HBE had increased viral load as compared to controls (103-104 copy/μL, P <0.001, N=4), and 2 to 3-fold increase in viral infection, respectively. TUNEL staining was increased in infected cells, indicating apoptosis. Transcript analysis of HBE cells with and without SARS-CoV-2 by RNASeq to identify differentially expressed genes in CSE exposed cells as compared to controls is in progress. Conclusion: Cigarette smoke and CSE increased ACE2 expression in ferrets, and ferret and human cells respectively. CSE-induction increased viral replication and infection severity, resulting in increased apoptosis. Cigarette smoking likely influences the severity of SARS-CoV-2 infection by altering expression of ACE2, inducing airway cell apoptosis upon infection.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"17 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121348112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Circulating Neutrophils from Critically Ill COVID-19 Patients Have Increased NETosis Via Canonical and Non-Canonical Pathways","authors":"J. Olay, J. Silva, A. Moshensky, A. Patel, M. Lam, M. Odish, E. Hansen, S. Trescott, C. Nguyen, R. Kim, K. Perofsky, S. Perera, L. J. Ma, J. Pham, M. Rolfsen, J. Shin, A. Fuentes, N. Coufal, A. Meier, L. Alexander","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1295","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1295","url":null,"abstract":"Introduction. The pathophysiology of infection with SARS-CoV-2 involves the lower airways and host-launched aggressive inflammatory responses leading to exacerbated lung damage in these vital tissues. Early clinical studies found that COVID-19 patients have higher levels of neutrophils in the circulation. Neutrophils are the most abundant leukocyte in circulation and are known to be highly proinflammatory due to production of neutrophil extracellular traps (NETosis). NETs are web-like chromatin structures coated with histones and proteases that both capture and kill invading pathogens. However, while being an effective countermeasure towards foreign microbes, this process also causes undesirable damage in host tissues. Therefore, we sought to characterize NETosis in circulating neutrophils from COVID-19 patients to determine whether this immunological response might be exacerbating or driving the disease state in COVID-19, rather than mitigating the virus. Methods.Blood was drawn daily from critically ill COVID-19 patients (n=16) after consent was obtained. Healthy controls (n=13) were screened for COVID-19 and gave blood once a week. Blood was drawn into lithium heparin tubes (BD Vacutainer). Neutrophils were isolated using PolymorphprepTM(PROGEN) per manufacturer's instructions. Cells were resuspended at 2x106 cells/ml for functional assays. Neutrophils were stimulated with increasing concentrations of PMA (Phorbol 12-myristate 13-acetate) of 2.5nM, 25nM and 250nM to stimulate NETosis via the canonical pathway, and nigericin at 15uM for the non-canonical pathway. NETosis was quantified using the Quant-iT™ PicoGreen™ dsDNA Assay Kit (Invitrogen) and by NET visualization via myeloperoxidase and nuclear staining (using Polyclonal Rabbit Anti-Human Myeloperoxidase by Dako and Hoescht stain by Invitrogen). Results.Functional NETosis assays of circulating neutrophils from COVID-19 patients demonstrate overall increased NETosis determined by increased release of dsDNA. This enhanced NETosis occurred at baseline and after stimulation with PMA when compared to healthy controls (Figure 1A, p <0.0001). Fluorescent microscopy also demonstrated increased NETosis in neutrophils from COVID-19 patients (Figure 1B;MPO-green and nucleus-blue). NETosis via the non-canonical pathway (induction with nigericin) was also increased in COVID-19 patients versus controls (p=0.02). Conclusions.Circulating neutrophils from critically ill COVID-19 patients are more prone to produce NETs than circulating neutrophils from healthy individuals. This is likely to lead to NETmediated tissue injury once neutrophils enter inflamed tissue, where they can potentially drive acute lung injury and acute respiratory distress syndrome, common causes of mortality in COVID-19. The finding of increased production of NETs by both canonical and non-canonical pathways is consistent with an overall hyper-activated state in COVID-19.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"97 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126828337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Interferon-Inducible Mediators Are Associated with SARS-CoV-2 and Related Severity Amongst Critically Ill Patients Suspected of COVID-19","authors":"E. Morrell, P. Bhatraju, N. Sathe, J. Lawson, A. Wiedeman, C. Vega, T. Liu, Xin-Ya Chai, S. Sahi, C. Brager, M. Orlov, S. Sakr, A. Sader, D. Lum, N. Koetje, A. Garay, E. Barnes, G. Cromer, M. Bray, S. Pipavath, S. Fink, Laura E. Evans, A. Long, T. West, M. Wurfel, C. Mikacenic","doi":"10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1294","DOIUrl":"https://doi.org/10.1164/AJRCCM-CONFERENCE.2021.203.1_MEETINGABSTRACTS.A1294","url":null,"abstract":"RATIONALE: Several studies have identified host immune signatures that are associated with COVID-19. However, it is unclear whether these immune signatures are specific to COVID-19 or are merely reflective of illness severity. In vitro studies have demonstrated that human T-cell responses to SARS-CoV-2-specific antigens are mediated through interferon-gamma (IFN-γ). Methods: We prospectively enrolled a multi-site cohort of patients admitted to the ICU under suspicion for COVID-19 who were then determined to be SARS-CoV-2-positive (n = 82) or-negative (n = 97) by RT-PCR. We measured multiple molecular and cellular immune profiles from blood and endotracheal aspirates (ETAs) collected on ICU admission. Our primary analysis tested for associations between IFN-γ and interferon-inducible mediators (CXCL10 and soluble PD-L1 (sPD-L1)) in blood or ETAs and SARS-CoV-2 status. We then stratified our cohort into SARS-CoV-2-negative and-positive groups and tested for associations between interferon-inducible mediators and clinical outcomes and SARS-CoV-2-copy-number. We used cytometry time-of-flight (CyTOF) to simultaneously measure 39 cell surface and intracellular markers on peripheral blood mononuclear cells collected from a subset of patients with ARDS. We then compared immune cell signatures in subjects with vs. without SARS-CoV-2. Results: The mean APACHE III score was higher in SARS-CoV-2-negative vs.-positive subjects (80±30 vs. 69±29), but the groups were otherwise well-matched. SARS-CoV-2-positive subjects had higher plasma concentrations of IFN-γ, CXCL10, and sPD-L1 relative to SARSCoV-2-negative patients adjusting for age, sex, and severity of illness (all p ≤ 0.01). The levels of IL-6, TNF-α, IL-8, MCP-1, and IL-17A were not significantly different between the two groups. SARS-CoV-2-positive subjects also had higher CXCL10 concentrations in ETAs than SARS-CoV-2-negative subjects. Higher plasma concentrations of CXCL10 and sPD-L1 were associated with higher mortality (Table 1) and more severe respiratory disease (ventilator-free days (VFDs), ARDS) in SARS-CoV-2-positive, but not-negative, patients. In contrast, higher IL-6 was associated with a lower number of VFDs and ARDS in both groups. IFN-γ and CXCL10 (but not IL-6) were associated with SARS-CoV-2-copy-number. Using CyTOF, we found SARS-CoV-2-positive subjects had a lower proportion of CXCR3+ (CXCL10 receptor) T-cells, a higher proportion of PD-L1+ monocytes, and less T-cell and monocyte intracellular cytokine staining vs. SARS-CoV-2-negative patients. Conclusion: Our findings suggest interferon-inducible mediator responses and immune cell hypofunction are characteristic of critically ill subjects with SARS-CoV-2 compared with similar patients without SARS-CoV-2. Our identification of immune signatures that are associated with SARS-CoV-2 infection but are distinct from other forms of critical illness clarifies COVID-19 pathophysiology.","PeriodicalId":320542,"journal":{"name":"TP3. TP003 COVID-19 INFECTIONS, MECHANISMS, AND CLINICAL IMPLICATIONS","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134148838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}