13C-metabolic flux analysis of human adenovirus infection: Implications for viral vector production

Nuno Carinhas, Alexey Koshkin, Daniel A.M. Pais, Paula M. Alves, Ana P. Teixeira

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Adenoviruses are human pathogens increasingly used as gene therapy and vaccination vectors. However, their impact on cell metabolism is poorly characterized. We performed carbon labeling experiments with [1,2-13C]glucose or [U-13C]glutamine to evaluate metabolic alterations in the amniocyte-derived, E1-transformed 1G3 cell line during production of a human adenovirus type 5 vector (AdV5). Nonstationary 13C-metabolic flux analysis revealed increased fluxes of glycolysis (17%) and markedly PPP (over fourfold) and cytosolic AcCoA formation (nearly twofold) following infection of growing cells. Interestingly, infection of growth-arrested cells increased overall carbon flow even more, including glutamine anaplerosis and TCA cycle activity (both over 1.5-fold), but was unable to stimulate the PPP and was associated with a steep drop in AdV5 replication (almost 80%). Our results underscore the importance of nucleic and fatty acid biosynthesis for adenovirus replication. Overall, we portray a metabolic blueprint of human adenovirus infection, highlighting similarities with other viruses and cancer, and suggest strategies to improve AdV5 production. Biotechnol. Bioeng. 2017;114: 195–207.

Original languageEnglish
Pages (from-to)195-207
Number of pages13
JournalBiotechnology and Bioengineering
Volume114
Issue number1
DOIs
Publication statusPublished - 1 Jan 2017

Keywords

  • C-metabolic flux analysis
  • adenovirus infection
  • mammalian cell culture
  • viral vector production

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