Increased dose and duration of infection with human adenovirus serotype 5 shifts amino acid, nucleic acid, TCA, and carbohydrate metabolism in human kidney cells

Abstract

Viruses are obligate intracellular parasites that overtake host cellular processes and establish an environment optimal for replication. Human adenoviruses are double stranded DNA viruses known to possess a unique tissue tropism including respiratory, gastrointestinal, ocular, and renal tissues. Furthermore, infection can result in mild to moderate symptoms with increased severity in those with dampened immunity. Disruptions to host metabolism is a well-known consequence of viral infection. Nonetheless, the relationship between adenoviruses and host metabolism remains a largely unelucidated area. Few studies have coupled metabolomic techniques with the examination of infection across the adenoviruses’ range of tissue targets. Fewer studies have investigated metabolic shifts with manipulation of viral dosage and duration of infection simultaneously. The objective for this study was to determine metabolic shifts in human kidney cells with different dosages of human adenovirus serotype 5 and couple this to duration of infection. It was hypothesized that prolonged viral infection with increased viral dosage would result in greater shifts in metabolites involved in amino acid metabolism, carbohydrate metabolism, nucleotide metabolism, and the tricarboxylic acid cycle. Three dosages of human adenovirus serotype 5 were used: 0.5. 1.0, and 2.0 multiplicity of infection. Infections were prolonged for 6, 12, 24, and 36 hours and cytosolic fractions were subsequently collected. Samples were then subjected to gas chromatography coupled to time-of-flight mass spectrometry. We found distinct separations in many primary carbon classes between uninfected controls and infected cells. Greater separation was seen in the higher dosed samples compared to the 0.5 MOI group. Analyses revealed 186 identified metabolites with significant upregulated and downregulated metabolites involved in nucleotide biosynthesis, amino acid metabolism, carbohydrate formation, and tricarboxylic acid cycle. Metabolites such as urea and pyrophosphate displayed the greatest degree of upregulation and downregulation, respectively. This was evident across all concentrations at 24 HPI. An increase in duration of infection was correlated with the emergence of more metabolites, with greater detection at increased viral concentration. Our results identify specific metabolic pathways perturbed by viral-host interactions, which have not been previously implicated in adenoviral infection and in kidney cells. Our study also provides a framework to investigate individual adenoviral protein contribution to metabolic disruptions. UC Merced USDA Hispanic Education Training Program Grant This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.