In a recent study published on the medRxiv * prepress server, researchers conducted an integrative analysis of clinical outcomes and viral entry networks in severe coronavirus disease infection 2019 (COVID-19).
Study: Integrative analysis of viral entry networks and clinical outcomes identifies a protective role of spironolactone in severe COVID-19. Image credit: Irina Anosova / Shutterstock
Fund
Viral entry into the host cell is a critical step in the coronavirus 2 (SARS-CoV-2) replication cycle of severe acute respiratory syndrome responsible for virulence and tropism of viral variants. The entry of SARS-CoV-2 into the host depends primarily on the binding interactions between the host angiotensin-2 converting enzyme (ACE-2) and the SARS-CoV-2 ear protein. . However, research is needed to understand the entire process of host cell entry and the involvement of the network of various host genes.
About the study
In the present study, researchers used high-performance functional screens and retrospective cohort analyzes to recognize drugs that can alter the course of COVID-19 disease by targeting factors influencing host entry.
The team recognized the host subnets that allow the entry of SARS-CoV-2 by collecting all short palindromic repetition (CRISPR) screens clustered throughout the genome. These screens estimated the effect of individual gene expression on SARS-CoV-2 infection. They provided data related to various cellular contexts, such as lung and non-lung cell types, as well as functional disturbances such as loss of function or CRISPR-KO and gain of function or CRISPRa. The final data set consisted of three CRISPRa and five CRISPR-KO screens that allowed the resolution of viral entry mechanisms.
The team also quantified the enrichment of the functional pathways between each CRISPR display. In addition, unweighted networks represented known interactions between Food and Drug Administration (FDA) approved drugs and input genes detected on individual screens. Subsequently, the team performed downstream analyzes of the drugs according to the centrality of each successful network.
In addition, a retrospective clinical analysis was performed with matching propensity scores to estimate whether the use of the candidate drugs was related to the severity of COVID-19 disease. The team collected electronic medical records, including a total of 64,349 cases from patients with a positive diagnosis of SARS-CoV-2. In addition, the team evaluated whether inhibition of SARS-CoV-2 entry could interfere with the mechanism of spironolactone. This was achieved by performing a SARS-CoV-2 pseudoviral entry assay at different doses of spironolactone on a cell line derived from the human pulmonary epithelium.
Results
The results of the study showed that the three CRISPRa and five CRISPR-KO screens showed different levels of correlation at single-gene levels consistent with the corresponding heterogeneous cell contexts. Nearly 88% of the screams were substantially correlated with a minimum of another CRISPR display, and 26% of the pairwise comparisons showed a noticeable positive correlation after adjusting for multiple tests. The team also noted that gene-level similarities were higher between each CRISPR screen type. A positive correlation was found between 60% and 33% of the CRISPR-KO and CRISPRa screen pairs, respectively.
The team also observed that a total of 20 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were substantially enriched in a minimum of two screens, including several pathways that were directly involved in SARS entry. -CoV-2. The pathways related to phosphoglycerides and glycosaminoglycans were the most enriched, according to their important function in viral binding. In addition, significant enrichment was also observed in pathways related to neurodegenerative diseases such as Huntington’s, Parkinson’s and Alzheimer’s. The team also found that the correlations between the enrichment scores of the normalized pathways for each screen were higher than those of the individual genes.
The study showed that each unweighted network comprised an average number of 116 genes, 605 drugs and 758 edges each, which corresponded to an average density of almost 1.06%. For each graph, the mean grade was estimated to be 6.50 for the genetic nodes, 1.23 for the drug nodes, and 2.03 in general, with approximately 3.81% of the genes detected per screen with unknown pharmacological interactions.
Spironolactone analysis revealed a substantial negative correlation between spironolactone use and progression of COVID-19 disease to admission to the intensive care unit (ICU). In addition, the association between spironolactone use and the need for a mechanical ventilator was significantly negative. Metformin, the positive control, also showed a marked negative correlation with ICU admission and a nominally substantial correlation with intervention with a mechanical ventilator.
The team observed a dose- and time-dependent effect of spironolactone at SARS-CoV-2 entry. Interestingly, there was an initial peak of SARS-CoV-2 entry between four and eight hours after infection when there was a slight but substantial increase in viral infection at the highest dose administration. After the initial peak, the team observed a larger and more robust reduction in viral infection at higher doses of spironolactone, which coincided with a global inhibition of SARS-CoV-2 entry.
Overall, the study’s findings showed that epironolactone could serve as a potential candidate as a modulator of SARS-CoV-2 infection.
* Important news
medRxiv publishes preliminary scientific reports that are not peer-reviewed and therefore should not be considered conclusive, guided by clinical practice or health-related behavior, or treated as established information.