13 C-metabolic flux analysis of human adenovirus infection: Implications for viral vector production.
Biotechnol Bioeng
; 114(1): 195-207, 2017 01.
Article
em En
| MEDLINE
| ID: mdl-27477740
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.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cultura de Vírus
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Isótopos de Carbono
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Adenoviridae
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Infecções por Adenovirus Humanos
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Análise do Fluxo Metabólico
Limite:
Humans
Idioma:
En
Revista:
Biotechnol Bioeng
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Portugal