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13 C‐metabolic flux analysis of human adenovirus infection: Implications for viral vector production
Author(s) -
Carinhas Nuno,
Koshkin Alexey,
Pais Daniel A.M.,
Alves Paula M.,
Teixeira Ana P.
Publication year - 2017
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26063
Subject(s) - metabolic flux analysis , glutamine , biology , metabolic engineering , glycolysis , cell culture , adenoviridae , flux (metallurgy) , citric acid cycle , metabolic pathway , virology , viral vector , metabolism , cell cycle , viral replication , virus , biochemistry , cell , gene , genetic enhancement , amino acid , chemistry , recombinant dna , genetics , organic chemistry
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‐ 13 C]glucose or [U‐ 13 C]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 13 C‐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. © 2016 Wiley Periodicals, Inc.