Metabolic Profiling from an Asymptomatic Ferret Model of SARS-CoV-2 Infection

Abstract

Coronavirus disease (COVID-19) is a contagious respiratory disease that is causing significant global morbidity and mortality. Understanding the impact of the Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) infection on the host metabolism is still in its infancy but of great importance. Herein, we investigated the metabolic response during viral shedding and post-shedding in an asymptomatic SARS-CoV-2 ferret model (n = 6) challenged with two SARS-CoV-2 isolates. Virological and metabolic analyses were performed on (minimally invasive) collected oral swabs, rectal swabs, and nasal washes. Fragments of SARS-CoV-2 RNA were only found in the nasal wash samples in four of the six ferrets, and in the samples collected 3 to 9 days post-infection (referred to as viral shedding). Central carbon metabolism metabolites were analyzed during viral shedding and post-shedding periods using a dynamic Multiple Reaction Monitoring (dMRM) database and method. Subsequent untargeted metabolomics and lipidomics of the same samples were performed using a Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry (LC-QToF-MS) methodology, building upon the identified differentiated central carbon metabolism metabolites. Multivariate analysis of the acquired data identified 29 significant metabolites and three lipids that were subjected to pathway enrichment and impact analysis. The presence of viral shedding coincided with the challenge dose administered and significant changes in the citric acid cycle, purine metabolism, and pentose phosphate pathways, amongst others, in the host nasal wash samples. An elevated immune response in the host was also observed between the two isolates studied. These results support other metabolomic-based findings in clinical observational studies and indicate the utility of metabolomics applied to ferrets for further COVID-19 research that advances early diagnosis of asymptomatic and mild clinical COVID-19 infections, in addition to assessing the effectiveness of new or repurposed drug therapies.

Dissecting the Role of the Human Microbiome in COVID-19 via Metagenome-assembled Genomes

“…Moreover, our analysis enables the direct microbial genome comparison between permissive and protective nrMAGs. Consistent with previous reports of a relationship of the pentose phosphate pathway and SARS-CoV-2 infection 50-52 , our findings support that overrepresentation of permissive nrMAGs and underrepresentation of protective nrMAGs may upregulate the pentose phosphate pathway as their genome are shown to be highly intact in those relevant modules. In contrast, the pentose phosphate pathway related KEGG modules showed significant lower module completeness in genomes of protective nrMAGs.…”

“…Importantly, a previous study reported a significant increase in the levels of some intermediates of the glycolytic and pentose phosphate pathways in sera of COVID-19 positive patients 50 . Moreover, SARS-CoV-2 infection was found to be associated with changes in the regulation of the pentose phosphate pathway in both in vivo (Caco-2 cells) 51 and in vitro (ferret model) 52 studies. These findings provide evidence of potential metabolic pathways linking the gut microbes to the clinical manifestations of COVID-19.…”

Implication of COVID-19 on Erythrocytes Functionality: Red Blood Cell Biochemical Implications and Morpho-Functional Aspects

“…The metabolic shift towards ATP production in the glycolytic pathway clearly, also influences other metabolic pathways, such as the pentose phosphate pathway (PPP). In these conditions, the non-oxidative phase of PPP is increased, in which glycolysis intermediates are converted into the ribose-5-phosphate required for the synthesis of the nucleic acids, which is necessary for the virus replication [25,26]. In addition, high levels of oxidized glutathione and a significant decrease in enzymes forming part of the cellular defense line against oxidative stress were found, including superoxide dismutase 1 (SOD1), glucose-6-phosphate dehydrogenase, and peroxiredoxin [27][28][29].…”

Targeting the Pentose Phosphate Pathway for SARS-CoV-2 Therapy

“…The non-oxidative PPP branch converts glycolytic intermediates into ribose-5-phosphate, required for the synthesis of nucleic acids, as well as sugar phosphate precursors that are necessary for the synthesis of amino acids [ 14 ]. Moreover, SARS-CoV-2 infection was associated with changes in the PPP in a ferret model [ 15 ]. Hence, in this study, we investigated whether the anti-SARS-CoV-2 activity of 2DG can be increased by additionally targeting the non-oxidative PPP.…”