Metabonomic Profiling Displayed Differing Effects Of Two Borna Disease Viruses Infection In Rat Cortical Neurons

Background and ObjectiveBorna disease virus(BDV) persists in the central nervous systems of a wide variety of vertebrates and causes behavioral disorders. Previous studies have revealed that metabolic perturbations are associated with BDV infection. However, the pathophysiological effects of different viral strains remain largely unknown. Strain V deprived of equine serving as the reference virus; while Hu-H1 deprived of human but its authenticity of human strains has been questioned simply based on the high level of sequence conservation of BDV(>95%) in very short gene stretches instead of complete genome sequencesThe aim of this study was to compare BDV infection with human strain Hu-H1 and lab reference strain Strain V differentially alters metabolic pathways of rat cortical neurons in vitro and to provided strong evidence for the authenticity of Hu-H1.MethodsRat cortical neurons infected with human strain BDV Hu-H1, laboratory BDV Strain V, and non-infected control(CON) cells were cultured in vitro. At day 12 post-infection, a gas chromatography coupled with mass spectrometry(GC-MS) metabonomic approach was used to differentiate the metabonomic profiles of 35 independent intracellular samples from Hu-H1-infected cells(n=11), Strain V-infected cells(n=12), and CON cells(n=12). Principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), and orthogonal partial least squares discriminant analysis(OPLS-DA) were performed to demonstrate discrimination between the three groups.ResultsWe identified 31 differential metabolites in the Hu-H1 and CON groups(21 decreased and 10 increased in Hu-H1 relative to CON), 35 differential metabolites in the Strain V and CON groups(30 decreased and 5 increased in Strain V relative to CON), and 21 differential metabolites in the Hu-H1 and Strain V groups(8 decreased and 13 increased in Hu-H1 relative to StrainV).ConclusionEmploying a GC-MS-based metabonomic approach, this is the first study to show that BDV infection with human strain Hu-H1 and lab reference strain Strain V differentially alters metabolic pathways of rat cortical neurons in vitro. Comparative metabonomic profiling revealed statistically significant perturbations in key energy, amino acid metabolites, cholesterol and fatty acid metabolites that were divergent between the two BDV strains with providing strong evidence for the authenticity of Hu-H1.This remarkably different effects of a natural human and a horse-derived cross-species adapted laboratory strain of BDV on neuronal cell metabolism found in our study also provides new avenues for future exploration of the pathogenic mechanism of BDV infection in vivo.