key: cord-1055008-731mdjyh authors: Zhang, M.; Du, L.; Faleti, O. D.; Huang, J.; Xiao, G.; Lyu, X. title: Higher pharyngeal epithelial gene expression of Angiotensin-Converting Enzyme-2 in upper respiratory infection patients date: 2020-06-29 journal: nan DOI: 10.1101/2020.06.27.20141754 sha: 9b4782eff9ed509f07943123ff4f4536423d2a90 doc_id: 1055008 cord_uid: 731mdjyh We analyzed the expression of ACE2 in pharyngeal epithelium and examined its relationship with clinical features and serological parameters in the upper respiratory infection (URI) patients. The expression of ACE2 were significantly higher in URI patients than in healthy controls individuals, and positively correlated with age and body temperature. We analyzed the expression of ACE2 in pharyngeal epithelium and examined its relationship with clinical features and serological parameters in the upper respiratory infection (URI) patients. The expression of ACE2 were significantly higher in URI patients than in healthy controls individuals, and positively correlated with age and body temperature. . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 29, 2020. . https://doi.org/10.1101/2020.06.27.20141754 doi: medRxiv preprint Co-infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and different respiratory pathogens has been reported recently [1] . It is hypothesized that the higher risk among coronavirus disease 2019 (COVID-19) patients with co-infection is due to differential expression of angiotensin-converting enzyme 2 (ACE2), the receptor that SARS-CoV-2 uses for host entry. We analyzed ACE2 gene expression in the pharyngeal epithelium of upper respiratory infection (URI) patients and healthy individuals. Real-time transcription-polymerase chain reaction analysis (RT-PCR) was used to determine expression of ACE2 gene. The relative expression level of ACE2 was normalized to the internal control GAPDH expression and calculated by the comparative C T (△△C T ) method. All data were statistically analyzed using Graph Pad Prism5 (version 5.0) software. Quantitative data are expressed as the mean±SD. Data with a Gaussian distribution was analyzed using an unpaired t test or one-way . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 29, 2020. . https://doi.org/10.1101/2020.06.27.20141754 doi: medRxiv preprint analysis of variance (ANOVA). Spearman's correlation test was performed to assess the correlation of ACE2 gene and clinical variables. P values less than 0.05 were considered statistically significant. Considering the evidence that ACE2 is the main host cell receptor of SARS-CoV-2 and plays a crucial role in the entry of virus into the cell to cause the final infection, ACE2 gene expression was the focus of this study. The cohort of 177 individuals aged 1 to 95 years, the median age of all URI patients was 33.69±15.88years, 57.6% were males (72/125). Among all URI patients, 65.6%(82/125) got a fever with body temperature over 37.4℃. The expression level of ACE2 gene was significantly higher in URI patients (n=125) than in Healthy control (HC) individuals (n=52) (p<0.0001, Fig.1) . ACE2 gene expression level was significantly and positively correlated with age (r=0.1799, p=0.0447) and body temperature (r=0.1927, p=0.0427) ( Table 1) . . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 29, 2020. . https://doi.org/10.1101/2020.06.27.20141754 doi: medRxiv preprint Data are means and 95% confidence intervals (error bars) for ACE2 gene expression in the pharyngeal epithelium of URI patients and HC individuals, *** (p<0.0001). Data were log-transformed by taking the base 10 logarithm to account for the skewness. After the above processing, the treated data were normal distributed and had constant variances. . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 29, 2020. In this study, we showed that ACE2 gene level was significantly upregulated in the pharyngeal epithelium of URI patients. We also found that ACE2 gene level was significantly and positively correlated with age and body temperature. So far as we know, there is few reports demonstrating that ACE2 gene level in the pharyngeal epithelium is increased in URI patients. A recent study reported ACE2 is an interferon-stimulated gene in human airway epithelial cells, which may help explain increasing coinfections with SARS-CoV-2 and other respiratory pathogens (2) . The Chinese experts from China-Japan Friendship Hospital have reported a case coinfection with influenza A virus and SARS-CoV-2 [3] . A recent study also showed that a total of 5 of 115 patients confirmed with COVID-19 was diagnosed with . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 29, 2020. . https://doi.org/10.1101/2020.06.27.20141754 doi: medRxiv preprint influenza virus infection [4] . There are still other case reports have described coinfection with other respiratory pathogens [1] . Those raise the concerns that there might be mixed infections of seasonal influenza and the novel coronavirus. Measures should be taken to enhance the respiratory infectious diseases surveillance systems and avoid lethal secondary infections. This study provides novel results on ACE2 gene expression in the pharyngeal epithelium and its relationship with URI disease. The molecular mechanism that how to activate ACE2 in URI patients might need to be future explored. Funding: This work was supported by grants from National Natural Science Foundation of The Importance of Novel Coronavirus Disease (COVID-19) and Coinfection with Other Respiratory Pathogens in the Current Pandemic SARS-CoV-2 Receptor ACE2 Is an Interferon-Stimulated Gene in Human Airway Epithelial Cells and Is Detected in Specific Cell Subsets across Tissues Co-infection with SARS-CoV-2 and Influenza A Virus in Patient with Pneumonia The clinical characteristics of pneumonia patients coinfected with 2019 novel coronavirus and influenza virus in Wuhan International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted June 29, 2020. . https://doi.org/10.1101/2020.06.27.20141754 doi: medRxiv preprint Potential conflicts of interest: None reported.The copyright holder for this preprint this version posted June 29, 2020. . https://doi.org/10.1101/2020.06.27.20141754 doi: medRxiv preprint