Metabolic reprogramming as a novel therapeutic target for Coxsackievirus B3

Figures

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  Figure 1.. CVB3 induces metabolic alteration through an increase in glycolysis. (A) Hela cells were infected with CVB3 for 12 h or 24 h. Then, plaque assays were then performed to quantify CVB3 titers. Means ± S.D., N = 3. (B) Measurement of the extracellular acidification rate (ECAR) (black line: post infection 0 h, blue line: post infection 12 h, and red line: post infection 24 h) (**P < 0.01, two-way ANOVA followed by Bonferroni’s post test). Means ± S.D., N = 3. (C) Flow cytometric analysis of mitochondrial membrane potential using JC-1 (**P < 0.01, Student’s t-test). Means ± S.D., N = 3.

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  Figure 2.. Confirmation of CVB3-mediated metabolic changes through metabolite analysis. Principal component analysis (PCA) score plots (A) and loading plots (B) derived from 23 metabolites. When metabolites belonging to PC1 were classified according to similarity, fructose, leucine, isoleucine, valine, phenylalanine, glucose, glycerol and threonine were classified as one cluster (red rectangle). Moreover, regarding to PC2, aspartic acid, malic acid, phosphoric acid, fumaric acid, β-alanine, alanine, proline and glycine were classified one cluster (blue rectangle).

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  Figure 3.. Confirmation of CVB3-mediated metabolic changes through metabolite analysis. (A) Correlation matrix of 23 metabolites. The intensity of the blue or red color in each square represents the Pearson's correlation coefficient. One group, shown mostly in blue, comprised 6 amino acids (proline, alanine, glycine, β-alanine, aspartic acid, glutamic acid) and 3 organic acids (phosphoric acid, malic acid, fumaric acid). The other group, shown mostly in red, contained 7 amino acids (glutamine, serine, leucine, phenylalanine, threonine, valine, isoleucine), 3 organic acids (lactic acid, succinic acid, citric acid), 2 sugars (fructose, glucose), 1 amine (ethanolamine) and 1 sugar alcohol (glycerol). (B) Box plot of metabolite with the most significant change after CVB3 infection (**P < 0.01, Student’s t-test). Means ± S.D., N = 3.

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  Figure 4.. Glycolysis inhibitors as novel therapeutic strategies for CVB3 treatment. (A) Quantification of CVB3 titers in Hela cells treated with medium with or without glucose (1 g/L) (**P < 0.01, Student’s t-test). Means ± S.D., N = 3. (B) Quantification of CVB3 titers in Hela cells treated with glycolysis inhibitors, 2-deoxy-D-glucose (2-DG) or sodium oxamate (SO) after fixing the glucose concentration in the medium to 1 g/L. (**P < 0.01, Student’s t-test). Means ± S.D., N = 3.