التفاصيل البيبلوغرافية
| العنوان: |
Transient vitamin B5 starving improves mammalian cell homeostasis and protein production. |
| المؤلفون: |
Pourcel, Lucille1 (AUTHOR) Lucille.Pourcel@sib.swiss, Buron, Flavien1 (AUTHOR), Garcia, Fanny1 (AUTHOR), Delaloix, Margaux-Sarah1 (AUTHOR), Le Fourn, Valérie2 (AUTHOR), Girod, Pierre-Alain2 (AUTHOR), Mermod, Nicolas1 (AUTHOR) |
| المصدر: |
Metabolic Engineering. Jul2020, Vol. 60, p77-86. 10p. |
| مصطلحات موضوعية: |
*PANTOTHENIC acid, *PEROXISOME proliferator-activated receptors, *RECOMBINANT proteins, *TRANSCRIPTION factors, *HOMEOSTASIS, *CHO cell, *BIOLOGICAL fitness, *LIPID metabolism |
| مستخلص: |
Maintaining a metabolic steady state is essential for an organism's fitness and survival when confronted with environmental stress, and metabolic imbalance can be reversed by exposing the organism to fasting. Here, we attempted to apply this physiological principle to mammalian cell cultures to improve cellular fitness and consequently their ability to express recombinant proteins. We showed that transient vitamin B5 deprivation, an essential cofactor of central cellular metabolism, can quickly and irreversibly affect mammalian cell growth and division. A selection method was designed that relies on mammalian cell dependence on vitamin B5 for energy production, using the co-expression of the B5 transporter SLC5A6 and a gene of interest. We demonstrated that vitamin B5 selection persistently activates peroxisome proliferator-activated receptors (PPAR), a family of transcription factors involved in energy homeostasis, thereby altering lipid metabolism, improving cell fitness and therapeutic protein production. Thus, stable PPAR activation may constitute a cellular memory of past deprivation state, providing increased resistance to further potential fasting events. In other words, our results imply that cultured cells, once exposed to metabolic starvation, may display an improved metabolic fitness as compared to non-exposed cells, allowing increased resistance to cellular stress. • A novel vitamin B5 metabolic selection approach yields mammalian cells highly and stably expressing a gene of interest. • B5 deprivation may leads to a fasting state in CHO cells and to the persistent activation of PPAR transcription factors. • PPARα overexpression combined with B5 starvation effectively improves recombinant protein production by recombinant cells. • Cultured cells exposed to such metabolic starvation display improved metabolic fitness and resistance to cellular stress. [ABSTRACT FROM AUTHOR] |
| قاعدة البيانات: |
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