Overexpression of transcription factor Foxa1 and target genes remediate therapeutic protein production bottlenecks in Chinese hamster ovary cells.

Details

Serval ID
serval:BIB_D3B3130E78A9
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Overexpression of transcription factor Foxa1 and target genes remediate therapeutic protein production bottlenecks in Chinese hamster ovary cells.
Journal
Biotechnology and bioengineering
Author(s)
Berger A., Le Fourn V., Masternak J., Regamey A., Bodenmann I., Girod P.A., Mermod N.
ISSN
1097-0290 (Electronic)
ISSN-L
0006-3592
Publication state
Published
Issued date
04/2020
Peer-reviewed
Oui
Volume
117
Number
4
Pages
1101-1116
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Despite extensive research conducted to increase protein production from Chinese hamster ovary (CHO) cells, cellular bottlenecks often remain, hindering high yields. In this study, a transcriptomic analysis led to the identification of 32 genes that are consistently upregulated in high producer clones and thus might mediate high productivity. Candidate genes were associated with functions such as signaling, protein folding, cytoskeleton organization, and cell survival. We focused on two engineering targets, Erp27, which binds unfolded proteins and the Erp57 disulfide isomerase in the endoplasmic reticulum, and Foxa1, a pioneering transcription factor involved in organ development. Erp27 moderate overexpression increased production of an easy-to-express antibody, whereas Erp27 and Erp57 co-overexpression increased cell density, viability, and the yield of difficult-to-express proteins. Foxa1 overexpression increased cell density, cell viability, and easy- and difficult-to-express protein yields, whereas it decreased reactive oxygen species late in fed-batch cultures. Foxa1 overexpression upregulated two other candidate genes that increased the production of difficult- and/or easy-to-express proteins, namely Ca3, involved in protecting cells from oxidative stress, and Tagap, involved in signaling and cytoskeleton remodeling. Overall, several genes allowing to overcome CHO cell bottlenecks were identified, including Foxa1, which mediated multiple favorable metabolic changes that improve therapeutic protein yields.
Keywords
CHO cells, Foxa1 transcription factor, cell viability, protein folding, recombinant protein production
Pubmed
Web of science
Open Access
Yes
Create date
23/01/2020 14:13
Last modification date
03/05/2020 7:01
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