Four new eudesmane-type and one new eremophilane-type sesquiterpenes from the whole plant of Carpesium abrotanoides L.

The extract of the whole plant of Carpesium abrotanoides L. yielded five new sesquiterpenes including four eudesmanes (1-4) and one eremophilane (5). The new compounds were characterized by spectroscopic analysis especially 1D and 2D NMR spectroscopy and HRESIMS data. Structurally, both compounds 1 and 2 were sesquiterpene epoxides and 2 owned an epoxy group at C-4/C-15 position to form a spiro skeleton. Compounds 4 and 5 were two sesquiterpenes without lactones and 5 possessed a carboxy group in the molecule. Additionally, all the isolated compounds were preliminarily evaluated for the inhibitory activity against SARS-CoV-2 main protease. As a result, compound 2 showed moderate activity with an IC50 value of 18.79 µM, while other compounds were devoid of noticeable activity (IC50 > 50 µM).

Viral shedding time and its influencing factors in different site samples of patients infected with Severe acute respiratory syndrome coronavirus 2 omicron BA.2 variant

Objective: To analyze the viral shedding time and its influencing factors in different site samples of patients infected with severe acute respiratory syndrome coronavirus 2 (SARA-CoV-2) omicron BA.2 variant. Methods: Real-time fluorescence polymerase chain reaction was used to detect SARS-CoV-2 nucleic acid in nasopharyngeal swab, sputum and anal swab from 217 patients with coronavirus disease 2019 (COVID-19) who were infected with severe acute respiratory syndrome coronavirus 2 omicron BA.2 variant confirmed by gene sequencing in The First Affiliated Hospital of Naval Medical University (Second Military Medical University). The differences of viral shedding time of different site samples were compared. Stratified analysis and multiple linear regression analysis were used to explore the influencing factors of viral shedding time in different site samples. Results: The age of the 217 COVID-19 patients was 32.0 (24.0, 50.5) years old, 59.0% of them were males (n=128), and 41.0% were females (n=89). Eight (3.7%) cases were diagnosed with asymptomatic infection, 184 (84.8%) cases were mild type, 21 (9.7%) cases were moderate type, 3 (1.4%) cases were severe type, and 1 (0.5%) case was critical type. A total of 70 (32.3%) patients were treated with molnupiravir. The viral shedding time of SARS-CoV-2 nucleic acid in nasopharyngeal swab, sputum and anal swab was 13.0 (11.0, 17.0) d, 16.5 (13.0, 21.0) d and 10.0 (5.3, 11.0) d, respectively, with the differences being significant between them (all P<0.001). Age 60 years old, underlying diseases (especially hypertension, coronary artery diseases, or neurological diseases), and clinical classification of moderate type were risk factors for prolonged viral shedding time in nasopharyngeal swab;male sex and underlying diseases were risk factors for prolonged viral shedding time in sputum;and male sex was a risk factor for prolonged viral shedding time in anal swab. Multiple linear regression analysis showed that critical type was an independent risk factor for prolonged viral shedding time in nasopharyngeal swab (P<0.05), and male sex and underlying diseases were independent risk factors for prolonged viral shedding time in sputum (both P<0.05). Conclusion: Among patients infected with omicron BA.2 variant, the viral shedding time in sputum is the longest and that in anal swab is the shortest. Male patients and/or patients with underlying diseases have longer viral shedding time in sputum.

Clinical significance of serum interleukin 6 in elderly patients infected with severe acute respiratory syndrome Coronavirus 2 Omicron variant and its correlation with underlying diseases

Objective: To investigate the clinical significance of serum interleukin 6 (IL-6) in elderly patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) omicron variant and its correlation with underlying diseases. Methods: A total of 22 elderly patients (> 80 years old) infected with omicron variant, who were admitted to Department of Infectious Diseases, The First Affiliated Hospital of Naval Medical University (Second Military Medical University) from Apr. to Jun. 2022 and tested positive for SARS-CoV-2 RNA, were included. The level of serum IL-6 was measured by flow cytometry, and the level of serum C reactive protein (CRP) was measured by immunonephelometry. Patients were divided into pneumonia group (16 cases) and non-pneumonia group (6 cases) according to the imaging examination results, and were divided into severe group (severe and critical type, 5 cases) and non-severe group (mild and normal type, 17 cases) according to the condition. Binary logistic regression model and receiver operating characteristic (ROC) curve were used to analyze the correlation between serum IL-6 and CRP levels and the severity of the disease and whether it would progress to pneumonia. Meanwhile, the relationships between underlying diseases and serum IL-6 level were explored. Results: Among the 22 patients, 6 were mild, 11 were normal, 3 were severe, and 2 were critical. The baseline serum IL-6 level in the pneumonia group was significantly higher than that in the non-pneumonia group ([20.16+or-12.36] pg/mL vs [5.42+or-1.57] pg/mL, P=0.009), and there was no significant difference in baseline serum CRP level between the 2 groups (P > 0.05). There were no significant differences in baseline serum IL-6 or CRP levels between the severe group and the non-severe group (both P > 0.05). Logistic regression analysis showed that the baseline serum IL-6 and CRP might be related to pneumonia after infection with omicron variant (odds ratio [OR]=2.407, 95% confidence interval [CI] 0.915-6.328;OR=1.030, 95% CI 0.952-1.114). ROC curve analysis showed that the area under curve values of serum IL-6 and CRP in predicting the progression to pneumonia were 0.969 (95% CI 0.900-1.000) and 0.656 (95% CI 0.380-0.932), respectively, with statistical significance (Z=2.154, P=0.030). There were no significant differences in the baseline serum IL-6 level or proportions of severe patients or pneumonia patients among patients with or without hypertension, diabetes mellitus, coronary heart disease, chronic kidney disease or chronic obstructive pulmonary disease (all P > 0.05). The baseline serum IL-6 levels of the omicron variant infected elderly patients with 1, 2, and 3 or more underlying diseases were 12.50 (9.15, 21.75), 23.55 (9.63, 50.10), and 10.90 (5.20, 18.88) pg/mL, respectively, with no statistical significance (P > 0.05). Conclusion: For omicron variant infected patients, serum IL-6 level is significantly increased in patients with pneumonia manifestations and is correlated with disease progression. Serum IL-6 level is of great guiding significance to judge disease progression and evaluate efficacy and prognosis of elderly coronavirus disease 2019 patients.

Recent advances in double-lumen tube malposition in thoracic surgery: A bibliometric analysis and narrative literature review.

Thoracic surgery has increased drastically in recent years, especially in light of the severe outbreak of the 2019 novel coronavirus disease (COVID-19). Routine "passive" chest computed tomography (CT) screening of inpatients detects some pulmonary diseases requiring thoracic surgeries timely. As an essential device for thoracic anesthesia, the double-lumen tube (DLT) is particularly important for anesthesia and surgery. With the continuous upgrading of the DLTs and the widespread use of fiberoptic bronchoscopy (FOB), the position of DLT in thoracic surgery is gradually becoming more stable and easier to observe or adjust. However, DLT malposition still occurs during transferring patients from a supine to the lateral position in thoracic surgery, which leads to lung isolation failure and hypoxemia during one-lung ventilation (OLV). Recently, some innovative DLTs or improved intervention methods have shown good results in reducing the incidence of DLT malposition. This review aims to summarize the recent studies of the incidence of left-sided DLT malposition, the reasons and effects of malposition, and summarize current methods for reducing DLT malposition and prospects for possible approaches. Meanwhile, we use bibliometric analysis to summarize the research trends and hot spots of the DLT research.

Disease Progression of Hospitalized Elderly Patients with Omicron BA.2 Treated with Molnupiravir.

INTRODUCTION: The efficacy of molnupiravir (MLN) on Omicron sublineages is limited. We investigated the effectiveness of MLN in older adults diagnosed with Omicron BA.2. METHODS: Data of elderly COVID-19 patients (over 60 years) admitted to Chinghai Hospital (Shanghai, China) from 26 March to 31 May 2022 were reviewed. Study outcomes were a composite of undetectable viral load (VL) and disease progression [all-cause mortality, initiation of oxygen supply through high-flow device or invasive mechanical ventilation (IMV), or intensive care unit (ICU) admission] and their individual outcomes. RESULTS: A total of 42 elderly patients were enrolled: 26 of them received MLN, 17 (40.5%) were males, the median age was 84 years, and 12 were fully vaccinated (31.0%). Among these elderly COVID-19 patients, five (11.90%) experienced obvious dyspnea or were transferred to ICU [three MLN users (11.5%) versus two non-MLN users (12.5%)]. Compared with no MLN use, MLN use was associated with rapid undetectable VL. At day 10, MLN users achieved significantly greater undetectable VL than non-MLN users. Adjusted analysis showed that elderly patients who received MLN were 7.584 times more likely to achieve undetectable VL at day 10 than non-MLN users. Overall, elderly patients experienced a median hospital stay of 13 days. Compared with patients receiving standard care (SC), the median hospital stay of MLN users was reduced by 2.5 days. CONCLUSION: Early initiation of MLN in elderly COVID-19 was associated with fast undetectable VL and short hospital stay.

Comparison of Augmented Reality-assisted and Instructor-assisted Cardiopulmonary Resuscitation: A Simulated Randomized Controlled Pilot Trial.

Background: A trained lay rescuer is the most important determinant of survival from sudden cardiac arrest. Augmented Reality (AR) device may represent a powerful instrument for CPR assistance and self-training especially during the COVID-19 pandemic. Methods: A prospective, parallel, 1:1 pilot randomized clinical trial was designed. An AR CPR app was developed and 28 participants were randomly allocated into AR-assisted group and instructor-assisted group. Acceptability, usability, and mean per minute/per cycle chest compression depth, rate and accuracy were measured. Results: The mean scores for acceptability and usability were all rated good in each group. Comparing real-time AR-assisted CPR to instructor-assisted CPR, the mean difference of compression depth was 0.18 (95% CI: -0.18-0.53) cm and rate was -1.58 (95% CI: -6.11-2.95) min-1. Comparing AR self-training to instructor training, the AR group was not significantly different between two groups regarding both compression depth, rate and accuracy (p > .05). Conclusion: We found that the AR CPR app was an acceptable and usable tool both in real-time-assisted CPR and self-training CPR.

Development and Validation of a High-Throughput Next-Generation Sequencing Workflow for SARS-CoV-2 Whole Genome Sequencing: Results from Over 65,000 Clinical Cases (preprint)

Monitoring new mutations in SARS-CoV-2 provides crucial information for identifying diagnostic and therapeutic targets and important insights to achieve a more effective COVID-19 control strategy. Next generation sequencing (NGS) technologies have been widely used for whole genome sequencing of SARS-CoV-2. While various NGS methods have been reported, one chief limitation has been the complexity of the workflow, limiting the scalability. Here, we overcome this limitation by designing a workflow optimized for high-throughput studies. The workflow utilizes modified ARTIC network v3 primers for SARS-CoV-2 whole genome amplification. NGS libraries were prepared by a 2-step PCR method, similar to a previously reported tailed PCR method, with further optimizations to improve amplicon balance, to minimize amplicon dropout for viral genomes harboring primer-binding site mutation(s), and to integrate robotic liquid handlers. Validation studies demonstrated that the optimized workflow can process up to 2,688 samples in a single sequencing run without compromising sensitivity and accuracy and with fewer amplicon dropout events compared to the standard ARTIC protocol. We additionally report results for over 65,000 SARS-CoV-2 whole genome sequences from clinical specimens collected in the United States between January and September of 2021, as part of an ongoing national genomics surveillance effort.

Hypersensitive detection of transcription factors by multiple amplification strategy based on molecular beacon

Detection and quantification of specific targets at ultralow concentrations are crucial in biotechnological applications and biomedical diagnostics. However, traditional detection methods are time-consuming and complex. In this study, a simple, sensitive, and specific fluorescence-based strategy to detect transcription factors (TFs) was developed by combining molecular beacon (MB) with exonuclease III (Exo III). The strategy was validated using nuclear factor-kappa B (NF-κB) p65 as a model case. In this assay, the TF concentration is determined by estimating the concentration of the nucleic acid released through the reactions of TFs with DNA-1/DNA-2 duplex probe and Exo III;thus, this method can overcome the limitations of the traditional methods. The nucleic acid is then cyclically amplified in the presence of MB and Exo III, and the sequence fragments containing the fluorescent group are released. This method is sensitive, expandable, and suitable for the direct detection of TFs in crude nuclear extracts of cancer cells. Furthermore, it can provide key test data for studies of coronavirus disease 2019-induced acute respiratory distress syndrome and related research.

Evolutionary dynamics of indels in SARS-CoV-2 spike glycoprotein (preprint)

SARS-CoV-2, responsible for the current COVID-19 pandemic that claimed over 4.2 million lives, belongs to a class of enveloped viruses that undergo quick evolutionary adjustments under selection pressure. Numerous variants have emerged in SARS-CoV-2 that are currently posing a serious challenge to the global vaccination effort and COVID-19 management. The evolutionary dynamics of this virus are only beginning to be explored. In this work, we have analysed 1.79 million spike glycoprotein sequences of SARS-CoV-2 and found that the virus is fine-tuning the spike with numerous amino acid insertions and deletions (indels). Indels seem to have a selective advantage as the proportions of sequences with indels were steadily increasing over time, currently at over 89%, with similar trends across countries/variants. There were as many as 420 unique indel positions and 447 unique combinations of indels. Despite their high frequency, indels resulted in only minimal alteration, including both gain and loss, of N-glycosylation sites. As indels and point mutations are positively correlated and sequences with indels have significantly more point mutations, they have implications in the context of evolutionary dynamics of the SARS-CoV-2 spike glycoprotein.

Perceived uncertainty stress and its predictors among residents in China during the COVID-19 pandemic.

The prevalence of and risk factors for uncertainty stress among residents during the COVID-19 pandemic remain unclear. An online cross-sectional survey was conducted to explore and identify the risk factors for high perceived uncertainty stress among the general public in China during the COVID-19 outbreak. Information about the respondents' socioeconomic characteristics, knowledge of and attitudes towards COVID-19, perceived uncertainty stress, social capital, anxiety, and depressive symptoms was collected and analysed. Among the 1205 respondents, 45.3% (546) reported a high level of uncertainty stress. Multiple linear regression analysis indicated that anxiety (ß=3.871,P<0.001) and depression symptoms (ß=2.458, P<0.001), family residence (in towns or rural areas) (ß=0.947, P<0.001), lack of support for local epidemic control strategies (ß=1.253, P<0.001), worry about the pandemic (ß=1.191, P<0.001), and symptoms of weakness among family members (ß=1.525, P=0.002) were positively associated with perceived uncertainty stress. Cognitive social capital (ß=-0.883, P<0.001) and social networks (ß=-0.726, P<0.001) were negatively, but social participation (ß=0.714, P<0.001) was positively associated with perceived uncertainty stress. Our findings identify factors associated with a higher level of uncertainty stress and should be helpful in the consideration of effective policies and interventions for uncertainty stress during the initial phases of public health emergencies.

Early Treatment Effect of Chinese Herbal Medicine Formula Huashibaidu Granule on Mild COVID-19 Patients in Fangcang Hospital: An Unblinded, Cluster-Randomized Clinical Trial (preprint)

BACKGROUND: Previous study suggested that Chinese Herbal Medicine (CHM) Formula Huashibaidu granule might shorten disease course of Corona Virus Disease 2019 (COVID-19) patients. Our research aims to investigate the early treatment effect of Huashibaidu granule in mild COVID-19 patients under well clinical management.METHODS: An unblended cluster-randomized clinical trial was conducted at the Dongxihu FangCang hospital. 2 cabins were randomly allocated to CHM or control group, with 204 randomly sampled mild COVID-19 patients in each cabin. All participants received a 7-day conventional treatment, and CHM group cabin used additional Huashibaidu granule 10g twice daily. Participants were followed up until they met clinical endpoint. The primary outcome was patient become worsening before clinical endpoint occurred. The secondary outcomes was discharge with cure before clinical endpoint occurred and relief of composite symptoms after 7 days treatment.FINDINGS: All 408 participants were followed up to meet clinical endpoint and included in statistical analysis. The baseline characteristics were comparable between 2 groups. The number of worsening patients in the CHM group was 5 (2.5%), and that in the control group was 16 (7.8%). There was a significant difference between groups (P=0.014). 8 foreseeable mild adverse events occurred without statistical difference between groups.INTERPRETATION: 7-day early treatment with Huashibaidu granule reduced worsening conversion of mild COVID-19 patients. Our study supports Huashibaidu Granule as an active option for early treatment of mild COVID-19 in similar medical locations with well management.TRIAL REGISTRATION: The Chinese Clinical Trial Registry: ChiCTR2000029763.FUNDING: This study was supported by “National Key R&D Program of China” (No.2020YFC0841500).DECLARATION OF INTERESTS: The authors guaranteed that there existed no competing interest in this paper.ETHICS APPROVAL STATEMENT: Ethics Review Committee of Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences Approval of Ethical Review Acceptance Number: S2020-001; Approval Number: P20001/PJ01.

Authorized, Clear and Timely Communication of Risk to Guide Public Perception and Action: Lessons of COVID-19 from China (preprint)

Backgrounds: This study examined the dynamic association between risk communication and public’s risk perception and action across the COVID-19 outbreak timeline in China.Methods: Publicly available information on COVID-19 was collected by Parehub tool for official channels (e.g. government websites and official media), and by Zhongyun Big Data Platform for mainstream Chinese social media (e.g. TikTok, Weibo, etc.). Information was examined by its release channels, content and release time. An online survey was conducted via WeChat across 33 provinces and municipalities of China to assess changes in participants’ risk perception and action against COVID-19. Information content and release-time trajectories were examined against the public’s risk perception and actions over time.Results: Altogether, 1,477 pieces of authorized information and 297,000 pieces of short videos on COVID-19 were retrieved from official channels and social media. Our online survey of 1,311 responds (25-60 years, 42% male) indicated that participants mainly relied on official channels to obtain information. Alongside the progress of the outbreak, there was a gradual rise in information quantity, publishing frequency, and content variation. Correspondingly, the public’s risk perception that “take it seriously" rose from 13% to 80%, 87.1% of who took “multiple actions” compared to 25.9% initially.Conclusions: Our findings indicated that insufficient information freely-accessible at the early stages of the outbreak may nevertheless lead to the lack of risk awareness and inadequate protective actions of the public. Given the current global situation of COVID-19, the study highlights authorized, transparent, and timely two-way risk communication is vital to guide public perception and actions. 

Problems and strategies for receiving overseas donations when public health emergency occurs: a case study of Wuhan Union Hospital / 中华医院管理杂志

Since the COVID-19 epidemic broke out in Wuhan, medical institutions in different parts the country have experienced personal protective equipment (PPE) shortages at different levels. The situation is especially severe in the coronavirus epicenter. Overseas donation is one of the effective methods to ease the shortage of protective equipment. Based on the practice of Union Hospital of Tongji Medical College of Huazhong University of Science and Technology, the article discussed the difficulties and risks for public hospitals to receive overseas donations, summarized experience and proposed suggestions which can be used as reference for other public hospitals and related departments.

Authorized, Clear and Timely Communication of Risk to Guide Public Perception and Action: Lessons of COVID-19 From China (preprint)

Backgrounds: This study examined the dynamic association between risk communication and public’s risk perception and action across the COVID-19 outbreak timeline in China.Methods: Publicly available information on COVID-19 was collected by Parehub tool for official channels (e.g. government websites and official media), and by Zhongyun Big Data Platform for mainstream Chinese social media (e.g. TikTok, Weibo, etc.). Information was examined by its release channels, content and release time. An online survey was conducted via WeChat across 33 provinces and municipalities of China to assess changes in participants’ risk perception and action against COVID-19. Information content and release-time trajectories were examined against the public’s risk perception and actions over time.Results: Altogether, 1,477 pieces of authorized information and 297,000 pieces of short videos on COVID-19 were retrieved from official channels and social media. Our online survey of 1,311 responds (25-60 years, 42% male) indicated that participants mainly relied on official channels to obtain information. Alongside the progress of the outbreak, there was a gradual rise in information quantity, publishing frequency, and content variation. Correspondingly, the public’s risk perception that “take it seriously" rose from 13% to 80%, 87.1% of who took “multiple actions” compared to 25.9% initially.Conclusions: Our findings indicated that insufficient information freely-accessible at the early stages of the outbreak may nevertheless lead to the lack of risk awareness and inadequate protective actions of the public. Given the current global situation of COVID-19, the study highlights authorized, transparent, and timely two-way risk communication is vital to guide public perception and actions.

Modeling COVID-19 with Human Pluripotent Stem Cell-Derived Cells Reveals Synergistic Effects of Anti-inflammatory Macrophages with ACE2 Inhibition Against SARS-CoV-2 (preprint)

Dysfunctional immune responses contribute critically to the progression of Coronavirus Disease-2019 (COVID-19) from mild to severe stages including fatality, with pro-inflammatory macrophages as one of the main mediators of lung hyper-inflammation. Therefore, there is an urgent need to better understand the interactions among SARS-CoV-2 permissive cells, macrophage, and the SARS-CoV-2 virus, thereby offering important insights into new therapeutic strategies.  Here, we used directed differentiation of human pluripotent stem cells (hPSCs) to establish a lung and macrophage co-culture system and model the host-pathogen interaction and immune response caused by SARS-CoV-2 infection. Among the hPSC-derived lung cells, alveolar type II and ciliated cells are the major cell populations expressing the viral receptor ACE2 and co-effector TMPRSS2, and both were highly permissive to viral infection. We found that alternatively polarized macrophages (M2) and classically polarized macrophages (M1) had similar inhibitory effects on SARS-CoV-2 infection. However, only M1 macrophages significantly up-regulated inflammatory factors including IL-6 and IL-18, inhibiting growth and enhancing apoptosis of lung cells. Inhibiting viral entry into target cells using an ACE2 blocking antibody enhanced the activity of M2 macrophages, resulting in nearly complete clearance of virus and protection of lung cells. These results suggest a potential therapeutic strategy, in that by blocking viral entrance to target cells while boosting anti-inflammatory action of macrophages at an early stage of infection, M2 macrophages can eliminate SARS-CoV-2, while sparing lung cells and suppressing the dysfunctional hyper-inflammatory response mediated by M1 macrophages.    

Cardiac Troponin I Associated with Poor Prognosis and Death Risk in 726 Severe and Critical COVID-19 Patients: A Retrospective Cohort Study (preprint)

Background: A few patients with coronavirus disease 2019 (COVID-19) may progress into irreparable outcomes. Early identification of patients with serious symptoms who may develop critical illness and even death is of considerable importance for personalizing treatment and balancing medical resources.Methods: In this retrospective study, demographic, clinical characteristics and laboratory tests from 726 patients with serious COVID-19 from Tongji Hospital (Wuhan, China) were analyzed. The standards for the serious type are guided by the Chinese management guideline for COVID-19. Patients were classified into critical group (174 cases) and severe group (552 cases) based on whether the composite endpoint was reached, and the former group was divided into the survivors (47 cases) and non-survivors (127 cases). Univariable and multivariable logistic regression and receiver operating characteristic (ROC) curve analysis were performed to investigate the risk factors associated with poor prognosis and mortality outcomes.Results: Male patients accounted for 62.1% and 51.6% in the critical group and severe group, with a median age of 68 and 65 years, respectively. Among critical cases there was a higher prevalence of chronic obstructive lung disease (p = 0.029) and chest distress (p = 0.040) than in severe cases. In the multivariable analysis, the risk factors associated with poor prognosis in severe cases were advanced age (p = 0.002), high respiratory rate (RR) (p < 0.0001), high lactate dehydrogenase (LDH) level (p = 0.021), high hypersensitive cardiac troponin I (hs-cTnI) level (p < 0.0001), and low platelet counts (p = 0.005) at admission. In the adjusted models, higher mortality outcomes in critical patients were associated with high hs-cTnI level (p = 0.037). By plotting ROC curves of different indices, hs-cTnI and LDH were found to be predictive factors for poor prognosis in patients with severe COVID-19.Conclusions: For the risk assessment of serious COVID-19 patients on admission, advanced age, high level of RR, LDH, hs-cTnI, and low platelet counts, constitute important risk factors for poor prognosis in severe cases, and the hs-cTnI level can be helpful in predicting fatal outcomes in critically ill patients.

Identification of Candidate COVID-19 Therapeutics using hPSC-derived Lung Organoids (preprint)

Summary ParagraphThe SARS-CoV-2 virus has caused already over 3.5 million COVID-19 cases and 250,000 deaths globally. There is an urgent need to create novel models to study SARS-CoV-2 using human disease-relevant cells to understand key features of virus biology and facilitate drug screening. As primary SARS-CoV-2 infection is respiratory-based, we developed a lung organoid model using human pluripotent stem cells (hPSCs) that could be adapted for drug screens. The lung organoids, particularly aveolar type II cells, express ACE2 and are permissive to SARS-CoV-2 infection. Transcriptomic analysis following SARS-CoV-2 infection revealed a robust induction of chemokines and cytokines with little type I/III interferon signaling, similar to that observed amongst human COVID-19 pulmonary infections. We performed a high throughput screen using hPSC-derived lung organoids and identified FDA-approved drug candidates, including imatinib and mycophenolic acid, as inhibitors of SARS-CoV-2 entry. Pre- or post-treatment with these drugs at physiologically relevant levels decreased SARS-CoV-2 infection of hPSC-derived lung organoids. Together, these data demonstrate that hPSC-derived lung cells infected by SARS-CoV-2 can model human COVID-19 disease and provide a valuable resource to screen for FDA-approved drugs that might be repurposed and should be considered for COVID-19 clinical trials.

Identification of Drugs Blocking SARS-CoV-2 Infection using Human Pluripotent Stem Cell-derived Colonic Organoids (preprint)

Summary ParagraphThe current COVID-19 pandemic is caused by SARS-coronavirus 2 (SARS-CoV-2). There are currently no therapeutic options for mitigating this disease due to lack of a vaccine and limited knowledge of SARS-CoV-2 biology. As a result, there is an urgent need to create new disease models to study SARS-CoV-2 biology and to screen for therapeutics using human disease-relevant tissues. COVID-19 patients typically present with respiratory symptoms including cough, dyspnea, and respiratory distress, but nearly 25% of patients have gastrointestinal indications including anorexia, diarrhea, vomiting, and abdominal pain. Moreover, these symptoms are associated with worse COVID-19 outcomes1. Here, we report using human pluripotent stem cell-derived colonic organoids (hPSC-COs) to explore the permissiveness of colonic cell types to SARS-CoV-2 infection. Single cell RNA-seq and immunostaining showed that the putative viral entry receptor ACE2 is expressed in multiple hESC-derived colonic cell types, but highly enriched in enterocytes. Multiple cell types in the COs can be infected by a SARS-CoV-2 pseudo-entry virus, which was further validated in vivo using a humanized mouse model. We used hPSC-derived COs in a high throughput platform to screen 1280 FDA-approved drugs against viral infection. Mycophenolic acid and quinacrine dihydrochloride were found to block the infection of SARS-CoV-2 pseudo-entry virus in COs both in vitro and in vivo, and confirmed to block infection of SARS-CoV-2 virus. This study established both in vitro and in vivo organoid models to investigate infection of SARS-CoV-2 disease-relevant human colonic cell types and identified drugs that blocks SARS-CoV-2 infection, suitable for rapid clinical testing.