Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of human neural stem cell culture after infection with HCMV reveals HCMV infection disrupts the self-renewal capacity of neural progenitor cells and inhibits neuronal differentiation

ABSTRACT: Our results suggest that HCMV infection disrupts the self-renewal capacity of NPCs and influences their differentiation. Whole genome expression analysis revealed many changes in cellular gene expression, including downregulation of genes pertinent to the neuronal lineage. Experiment Overall Design: For gene expression analysis, cells were either mock- or virus-infected. Mock-infectedÂ cells were harvested at 12 hpi, virus infected cells were harvested at 4, 12 and 24 hpi. Virus- and mock-infections were performed in duplicate, with each individual sample harvested separately, thereby acting as biological replicates for analysis.

ORGANISM(S): Homo sapiens

SUBMITTER: Philip Hitchins Schwartz

PROVIDER: E-GEOD-19345 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:To combat virus infections, which are major human killers, a deeper understanding of how viruses reprogram their hosts to create optimal production of progeny is needed. Most knowledge on the regulation of cellular gene expression during adenovirus infection is derived from studies of mRNA expression. Here, we investigated the changes in cellular protein expression during the late phase of adenovirus type 2 (Ad2) infection of primary human cells by stable isotope labeling in cell culture (SILAC) with subsequent liquid chromatography-high resolution tandem mass spectrometric (LC-MS/MS) analysis using a Q-Exactive Orbitrap instrument. Cells were in-depth evaluated 24 and 36 hours post infection (hpi) and two biological replicates were investigated using swapped labeling. In total, 2648 and 2394 proteins were quantified at 24 and 36 hpi, respectively. Among them, 659 and 645 were deregulated more than 1.6-fold at the two time points, and approximately 80% of them had sustained regulation from 24 to 36 hpi. For the deregulated genes the change in protein expression was compared with that of RNA expression using cDNA sequencing data from the same time points of infection. The comparison revealed a surprisingly low correlation (r = 0.3). Consistency between changes in RNA and protein expression was observed for approximately 30% of the genes, whereas a majority showed either divergent or opposite expression profiles. The deregulated proteins were subjected to pathway and biological function analysis using bioinformatics tools. A surprising observation was that proteins related to carbohydrate metabolism were up-regulated at the protein level but unchanged at the RNA level whereas histone proteins were down-regulated at the protein level but up-regulated at the RNA level. Our results show that regulation of cellular gene expression in the response to viral infection is more complex than hitherto believed and several examples of posttranscriptional regulation of gene expression were observed. The results contribute to our understanding of host cell gene regulation during infection at a deeper level.

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Transcription profiling of human neural stem cell culture after infection with HCMV reveals HCMV infection disrupts the self-renewal capacity of neural progenitor cells and inhibits neuronal differentiation

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