title: 13C-metabolic flux analysis of human adenovirus infection: Implications for viral vector production.
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). Nonstationary13C-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.
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Avenida da República, Oeiras, 2781-157, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, Oeiras, 2780-157, Portugal.
authors.forename: Nuno
authors.initials: N
authors.lastname: Carinhas
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Avenida da República, Oeiras, 2781-157, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, Oeiras, 2780-157, Portugal.
authors.forename: Alexey
authors.initials: A
authors.lastname: Koshkin
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Avenida da República, Oeiras, 2781-157, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, Oeiras, 2780-157, Portugal.
authors.forename: Daniel A M
authors.initials: DA
authors.lastname: Pais
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Avenida da República, Oeiras, 2781-157, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, Oeiras, 2780-157, Portugal.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Avenida da República, Oeiras, 2781-157, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, Oeiras, 2780-157, Portugal.
authors.forename: Ana P
authors.initials: AP
authors.lastname: Teixeira
mesh_terms: D000256:Adenoviridae / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000258:Adenovirus Infections, Human / Q000378:metabolism / Q000821:virology; D002247:Carbon Isotopes / Q000032:analysis / Q000378:metabolism*; D002460:Cell Line; D005973:Glutamine / Q000378:metabolism; D006801:Humans; D064688:Metabolic Flux Analysis / Q000379:methods*; D008954:Models, Biological; D014776:Virus Cultivation / Q000379:methods*
chemical_list: D002247:Carbon Isotopes; D005973:Glutamine
keywords: 13C-metabolic flux analysis; adenovirus infection; mammalian cell culture; viral vector production