Effect of COVID-19 during pregnancy: Studying the maternal and neonatal outcomes and assessing the placental changes related to SARS-CoV-2 (preprint)

Abstract: Background: Pregnant females affected with COVID-19 are reported to have poorer disease outcomes as compared to non-pregnant females of a similar age group. COVID-19 may lead to adverse changes in the placenta, which needs to be studied. Methods: This is a case series of 63 pregnant women hospitalized with COVID-19 from May 2020 to February 2021.The primary outcomes were maternal death or complications. Results: 63 women were studied. 83.3% of women were in the age group of 26 to 35 years. 33% women had associated comorbidities. 68.3% of women tested positive in their third trimester, 15.9% and 11% tested positive in their second and first trimesters respectively. 73% women had mild disease and 27% women required oxygen support. 3/63 women died. One woman in the second and two women in the third trimester died respectively. Histopathological examination in 13 placentae (of 19 placentae examined) were suggestive of maternal and fetal malperfusion. Conclusion: Pregnant COVID-19 women may develop disease-related as well as obstetric complications.

Remdesivir for the treatment of COVID-19 disease: A retrospective comparative study of patients treated with and without Remdesivir (preprint)

BackgroundRemdesivir (RDV) in coronavirus disease 2019 (COVID-19) has been found to be beneficial in patients with severe disease; however, its role in mild-moderate disease and its optimal timing need to be identified. ObjectiveTo assess the course of illness and final outcome in patients who received RDV at various stages of illness, and compare it to the non-RDV group. MethodsThis is a retrospective data analysis of 1262 COVID-19 patients hospitalized from May5, 2020 to August 31, 2020. The primary outcomes were progression to mechanical ventilation (MV) or death. Kaplan Meier survival analysis and log rank test were used for evaluating primary outcomes. Results398 patients comprised the RDV group and 260 patients comprised the non-RDV group. 2/3rd of patients were above 50 years of age in both the groups and 3/4th patients were male. Mortality rate was 5.8% in RDV group (10.4% in non-RDV group). Mortality rate was 3.6%, 4% and 16.7% when RDV was started within 5 days, 5 to 10 days and after 10 days of symptom onset respectively. Fewer patients in RDV group progressed to MV (4.0% v/s 8.2%). Earlier discharge occurred in RDV group. Use of supplemental oxygen was observed in 44.7% patients in RDV group (54.2% in non-RDV group). No significant adverse events were observed with RDV. Survival analysis showed that probability of event (death) was significant for patients with hypertension (HT) and/or diabetes mellitus (DM) in RDV group. ConclusionEarly initiation of RDV is associated with shorter hospital stay, lower mortality as well as reduced need for supplemental oxygen and mechanical ventilation.

Molecular Profiling of COVID-19 Autopsies Uncovers Novel Disease Mechanisms (preprint)

Background: Current understanding of COVID-19 pathophysiology is limited by disease heterogeneity, complexity, and a paucity of studies evaluating patient tissues with advanced molecular tools. Methods: Autopsy tissues from two COVID-19 patients, one of whom died after a month-long hospitalization with multi-organ involvement while the other died after a few days of respiratory symptoms, were evaluated using multi-scale RNASeq methods (bulk, single-nuclei, and spatial RNASeq next-generation sequencing) to provide unprecedented molecular resolution of COVID-19 induced damage. Findings: Comparison of infected/uninfected tissues revealed four major regulatory pathways. Effectors within these pathways could constitute novel therapeutic targets, including the complement receptor C3AR1, calcitonin-like receptor or decorin. Single-nuclei RNA sequencing of olfactory bulb and prefrontal cortex highlighted remarkable diversity of coronavirus receptors. Angiotensin I converting enzyme 2 was rarely expressed, while Basignin showed diffuse expression, and alanyl aminopeptidase was associated with vascular/mesenchymal cell types. Comparison of lung and lymph node tissues from patients with different symptomatology with Digital Spatial Profiling resulted in distinct molecular phenotypes. Interpretation: COVID-19 is a far more complex and heterogeneous disease than initially anticipated. Evaluation of COVID-19 rapid autopsy tissues with advanced molecular techniques can identify pathways and effectors at play in individual patients, measure the staggering diversity of receptors in specific brain areas and other well-defined tissue compartments at the single-cell level, and help dissect differences driving diverging clinical courses among patients. Extension of this approach to larger datasets will substantially advance the understanding of the mechanisms behind COVID-19 pathophysiology. Funding: No external funding was used in this study.

Evaluating the Efficacy of Tocilizumab in Moderate to Severe COVID-19 with Progressive Illness despite Steroids: Identifying the Optimal Timing of its Administration in C3G study (preprint)

Background High mortality has been described in coronavirus disease 2019 (COVID-19) with cytokine release syndrome (CRS). Tocilizumab (TCZ), an interleukin-6 (IL-6) receptor antagonist may be associated with improved outcomes in such patients; however, the subgroups of patients who benefit the most need to be identified. Objective To analyze the efficacy and optimal timing of administration of TCZ in moderate to severe COVID-19 with features of CRS, where the response to steroids was poor. Methods This is a retrospective study of 125 patients admitted between May 5 to July 31, 2020, in a tertiary care hospital in western India, with moderate to severe COVID-19 who were treated with TCZ along with steroids. The primary outcomes were the need for mechanical ventilation (MV) or death, and secondary outcomes were a decrease in oxygen requirement and inflammatory markers; the incidence of secondary infections, and renal or hepatic dysfunction. Kaplan Meier survival analysis and log rank test were used for evaluating primary outcomes. Secondary outcomes were analyzed using the Wilcoxon Signed-Rank test. Results Among 1081 patients admitted during the study period, 125 were administered TCZ (median age, 56 [95% CI 54 - 60] years; 100 [80%] male). The commonest symptoms were fever (96%), cough (64%), and dyspnea (48.8%). 78.4% patients had comorbidities (hypertension 51.2%, diabetes 43.2%, obesity 25.6% and chronic cardiac disease 13.6%). Of 117 patients who were treated with TCZ before requiring MV, 18.8% progressed to MV. Overall, 25% of the patients needed MV support. 65.3% of patients were discharged by day 14 after TCZ administration. Mortality was nil, 16.2%, 50%, and 62.5% in patients who received TCZ on room air, low flow oxygen, high flow nasal cannula (HFNC) and bilevel positive airway pressure (BiPAP), and MV respectively; overall 24.8% of patients died. Survival analysis showed no difference in outcome with respect to age and gender, while progression to MV showed a statistically significant reduction for the event death (90.9% of patients who progressed to MV died as compared to 6.3% who did not; log rank test with p < 0.0001). No adverse events were noticed. Conclusion Mortality was least in patients of COVID-19 with CRS who received TCZ while on low flow oxygen. When administered in the early hypoxemic phase, TCZ is associated with reduced mortality and decreased need for mechanical ventilation.

A Comparative COVID 19 Characterizations and Clinical Course Analysis between ICU and Non ICU Settings (preprint)

Objective: With COVID-19 pandemic severely affecting India and Ahmedabad city being one accounting for half COVID cases, objective was to determine disease course and severity of in patients at a COVID care hospital. Design: A Clinical trial registry of India registered observational study (CTRI/2020/05/025247). Setting: Certified COVID hospital located in Ahmedabad, Gujarat, India. Participants: 549 COVID positive patients hospitalized between 15 th May to 10 th August, 2020 and treated in ICU and non ICU settings. Main Outcome Measure: Comparative analysis of demographic, clinical characteristics, investigations, treatment, complications and outcome of COVID patients in ICU and non ICU settings. Results: Of the 549 hospitalized COVID positive patients, 159 were admitted in ICU during disease course while 390 had ward admissions. Overall median age was 52 (1-86) years. The ICU group was older (>65years), with associated comorbidities like hypertension and diabetes (p<0.001); higher proportion of males (79.25%); with dyspnea as a major clinical characteristic and consolidation in lungs as a major radiological finding as compared to ward patients. C - reactive protein, D-Dimer and Ferritin were higher in ICU patients. Overall 50% females depicted elevated Ferritin levels. Steriods(92.45%)and tocilizumab (69.18%) were more frequently used for ICU patients . Remdesivir was prescribed to both ICU and non ICU patients. Favirapir was also a line of treatment for 25% of ICU patients. Convalescent plasma therapy was given to 7 ICU patients. Complications like acute kidney injury (13.84%), shock (10.69 %), sepsis and encephalopathy were observed in ICU patients. Overall mortality rate was 5.47 % with higher mortality among males in comparison to females (p<0.0001). Conclusion: About 29% of overall patients required ICU admission that was commonly elderly males. Chances of ICU admission were higher with baselines comorbidities (1.5 times) and dyspnea (3.4 times) respectively. A multi-specialty COVID care team and updated treatment protocols improves outcomes.

Cytotoxic lymphocytes are dysregulated in multisystem inflammatory syndrome in children (preprint)

Multisystem inflammatory syndrome in children (MIS-C) presents with fever, inflammation and multiple organ involvement in individuals under 21 years following severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. To identify genes, pathways and cell types driving MIS-C, we sequenced the blood transcriptomes of MIS-C cases, pediatric cases of coronavirus disease 2019, and healthy controls. We define a MIS-C transcriptional signature partially shared with the transcriptional response to SARS-CoV-2 infection and with the signature of Kawasaki disease, a clinically similar condition. By projecting the MIS-C signature onto a co-expression network, we identified disease gene modules and found genes downregulated in MIS-C clustered in a module enriched for the transcriptional signatures of exhausted CD8+ T-cells and CD56dimCD57+ NK cells. Bayesian network analyses revealed nine key regulators of this module, including TBX21, a central coordinator of exhausted CD8+ T-cell differentiation. Together, these findings suggest dysregulated cytotoxic lymphocyte response to SARS-Cov-2 infection in MIS-C.

SARS-CoV-2 causes severe alveolar inflammation and barrier dysfunction (preprint)

Infections with SARS-CoV-2 lead to mild to severe coronavirus disease-19 (COVID-19) with systemic symptoms. Although the viral infection originates in the respiratory system, it is unclear how the virus can overcome the alveolar barrier, which is observed in severe COVID-19 disease courses. To elucidate the viral effects on the barrier integrity and immune reactions, we used mono-cell culture systems and a complex human alveolus-on-a-chip model composed of epithelial, endothelial, and mononuclear cells. Our data show that SARS-CoV-2 efficiently infected epithelial cells with high viral loads and inflammatory response, including the interferon expression. By contrast, the adjacent endothelial layer was no infected and did neither show productive virus replication or interferon release. With prolonged infection, both cell types are damaged, and the barrier function is deteriorated, allowing the viral particles to overbear. In our study, we demonstrate that although SARS-CoV-2 is dependent on the epithelium for efficient replication, the neighboring endothelial cells are affected, e.g., by the epithelial cytokine release, which results in the damage of the alveolar barrier function and viral dissemination.