Characterization of glutathione proteome in CHO cells and its relationship with productivity and cholesterol synthesis

Valentine Chevallier, Erwin M. Schoof, Laetitia Malphettes, Mikael R. Andersen, Christopher T. Workman*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Glutathione (GSH) plays a central role in the redox balance maintenance in mammalian cells. Previous studies of industrial CHO cell lines have demonstrated a relationship between GSH metabolism and clone productivity. However, a thorough investigation is required to understand this relationship and potentially highlight new targets for cell engineering. In this study, we have modulated the GSH intracellular content of an industrial cell line under bioprocess conditions in order to further elucidate the role of the GSH synthesis pathway. Two strategies were used: the variation of cystine supply and the direct inhibition of the GSH synthesis using buthionine sulfoximine (BSO). Over time of the bioprocess, a correlation between intracellular GSH and product titer has been observed. Analysis of metabolites uptake/secretion rates and proteome comparison between BSO-treated cells and non-treated cells has highlighted a slowdown of the TCA cycle leading to a secretion of lactate and alanine in the extracellular environment. Moreover, an adaptation of the glutathione related proteome has been observed with an up-regulation of the regulatory subunit of glutamate cysteine ligase and a down-regulation of a specific glutathione transferase subgroup, the Mu family. Surprisingly, the main impact of BSO treatment was observed on a global down-regulation of the cholesterol synthesis pathways. As cholesterol is required for protein secretion, it could be the missing piece of the puzzle to finally elucidate the link between GSH synthesis and productivity. This article is protected by copyright. All rights reserved.
Original languageEnglish
JournalBiotechnology and Bioengineering
Volume117
Issue number11
Pages (from-to)3448-3458
Number of pages11
ISSN0006-3592
DOIs
Publication statusPublished - 2020

Keywords

  • CHO cells
  • Gutathione
  • Buthionine sulfoximine
  • Cholesterol
  • Proteomic

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