中国仓鼠卵巢细胞
未折叠蛋白反应
XBP1型
内质网
组蛋白脱乙酰酶抑制剂
细胞生物学
化学
下调和上调
组蛋白脱乙酰基酶
细胞培养
生物化学
生物
组蛋白
RNA剪接
遗传学
核糖核酸
基因
作者
Kamal Prashad Segar,Vikas Chandrawanshi,Sarika Mehra
标识
DOI:10.1016/j.jbiosc.2017.05.005
摘要
Process engineering to improve product quality and titers is gaining importance at late-stage cell culture process development. Valproic acid, a US Food and Drug Administration-approved histone deacetylase (HDAC) inhibitor, has been shown to improve cell culture performance with higher productivities and minimal effect on the product quality. However, the wider physiological impact of valproic acid on recombinant cells has not been investigated till date. In this study, we investigate the role of unfolded protein response pathway when immunoglobulin G (IgG)-secreting Chinese hamster ovary (CHO) cells are treated with valproic acid, resulting in a 3-fold increase in product titers and productivity. It is found that cells undergo an early transient endoplasmic reticulum (ER) stress on treatment with valproic acid, and subsequently adapt to perform as high producers. Induction of chaperones through enhanced XBP1 splicing activity and ATF6 activation suggests an increase in protein processing activity in these cells. We show that in addition to the enhanced recombinant mRNA expression of IgG heavy chain and light chain, the activation of unfolded protein response (UPR) pathway is critical to the increase in productivity of cells on valproic acid treatment. Further, upregulation of the UPR pathway is not through HDAC inhibition alone. To our knowledge, this is the first attempt to arrive at a phenotype-genotype mechanistic understanding of how valproic acid treatment enhances productivity in recombinant CHO cells.
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